Camel Milk Resistome in Kuwait: Genotypic and Phenotypic Characterization.

Antimicrobial resistance (AMR) is one of the major global health and economic threats. There is growing concern about the emergence of AMR in food and the possibility of transmission of microorganisms possessing antibiotic resistance genes (ARGs) to the human gut microbiome. Shotgun sequencing and in vitro antimicrobial susceptibility testing were used in this study to provide a detailed characterization of the antibiotic resistance profile of bacteria and their ARGs in dromedary camel milk. Eight pooled camel milk samples, representative of multiple camels distributed in the Kuwait desert, were collected from retail stores and analyzed. The genotypic analysis showed the presence of ARGs that mediate resistance to 18 classes of antibiotics in camel milk, with the highest resistance to fluoroquinolones (12.48%) and disinfecting agents and antiseptics (9%). Furthermore, the results pointed out the possible transmission of the ARGs to other bacteria through mobile genetic elements. The in vitro antimicrobial susceptibility testing indicated that 80% of the isolates were resistant to different classes of antibiotics, with the highest resistance observed against three antibiotic classes: penicillin, tetracyclines, and carbapenems. Multidrug-resistant pathogens including Klebsiella pneumoniae, Escherichia coli, and Enterobacter hormaechei were also revealed. These findings emphasize the human health risks related to the handling and consumption of raw camel milk and highlight the necessity of improving the hygienic practices of farms and retail stores to control the prevalence of ARGs and their transmission.

Anthracyclines and the risk of arrhythmias: A systematic review and meta-analysis.

BACKGROUND: There have been controversial findings from recent studies regarding anthracyclines use and the subsequent risk of arrhythmias. This study aimed to evaluate the existing evidence of the risk of arrhythmias in patients treated with anthracyclines. METHODS: PubMed, Scopus, and Web of Science databases were searched up to April 2022 using keywords such as "anthracycline" and "arrhythmia." Dichotomous data were presented as relative risk (RR) and confidence interval (CI), while continuous data were presented as mean difference (MD) and CI. Revman software version 5.4 was used for the analysis. RESULTS: Thirteen studies were included with a total of 26891 subjects. Pooled analysis showed that anthracyclines therapy was significantly associated with a higher risk of arrhythmia (RR: 1.58; 95% CI: 1.41-1.76; P < .00001), ST segment and T wave abnormalities (RR: 1.73, 95% CI: 1.18-2.55, P = .005), conduction abnormalities and AV block (RR = 1.86, 95% CI = 1.06-3.25, P = .03), and tachycardia (RR: 1.736, 95% CI: 1.11-2.69, P = .02). Further analyses of the associations between anthracyclines and atrial flutter (RR = 1.30, 95% CI = 0.29-5.89, P = .74), atrial ectopic beats (RR: 1.27, 95% CI: 0.78-2.05, P = .34), and ventricular ectopic beats (RR: 0.93, 95% CI: 0.53-1.65, P = .81) showed no statistically significant results. Higher doses of anthracycline were associated with a higher risk of arrhythmias (RR: 1.49; 95% CI: 1.08-2.05; P = .02) compared to the lower doses (RR: 1.36; 95% CI: 1.00-1.85; P = .05). Newer generations of Anthracycline maintained the arrhythmogenic properties of previous generations, such as Doxorubicin. CONCLUSION: Anthracyclines therapy was significantly associated with an increased risk of arrhythmias. Accordingly, Patients treated with anthracyclines should be screened for ECG abnormalities and these drugs should be avoided in patients susceptible to arrhythmia. The potential benefit of the administration of prophylactic anti-fibrotic and anti-arrhythmic drugs should also be explored.

Draft genome sequence data of Enterococcus faecium R9, a multiple enterocins-producing strain.

Food contamination by pathogens results in serious health problems and economic losses. Chemical food preservatives pose a risk to human health when used in food preservation. To increase the shelf life of the products and prevent spoilage, the dairy sector is considering natural preservatives such the ribosomally synthesized peptides, bacteriocins. Here we present the draft genome sequence of Enterococcus faecium strain R9 producing three bacteriocins isolated from raw camel milk. These bacteriocins showed valuable technological properties, such as sensitivity to proteolytic enzymes, heat stability, and wide range of pH tolerance. The 2 × 250 bp paired end reads sequencing was performed on Illumina HiSeq 2500 sequencing. The genome sequence consisted of 3,598,862 bases, with a GC content of 37.94% bases. The number of raw reads was 4,670,510, and the assembly N50 score was 65,355 bp with a 310.28 average coverage. A total of 3,086 coding sequences (CDSs) was predicted with 2,126 CDSs with a known function and 127 with a signal peptide. Annotation of the genome sequence revealed bacteriocins encoding genes, namely, enterocin B, enterocin P, and two-component enterocin X (X-alfa and X-beta subunits). These enterocins are beneficial for controlling Listeria monocytogenes in the food industry. Genome sequence of Enterococcus faecium R9 has been deposited at the gene bank under BioSample accession number JALJED000000000 and are available in Mendeley Data [1].

Antibiotic Resistance Genes in Aerosols: Baseline from Kuwait.

Antimicrobial resistance (AMR) is one of the biggest threats to human health worldwide. The World Health Organization (WHO, Geneva, Switzerland) has launched the "One-Health" approach, which encourages assessment of antibiotic-resistant genes (ARGs) within environments shared by human-animals-plants-microbes to constrain and alleviate the development of AMR. Aerosols as a medium to disseminate ARGs, have received minimal attention. In the present study, we investigated the distribution and abundance of ARGs in indoor and outdoor aerosols collected from an urban location in Kuwait and the interior of three hospitals. The high throughput quantitative polymerase chain reaction (HT-qPCR) approach was used for this purpose. The results demonstrate the presence of aminoglycoside, beta-lactam, fluoroquinolone, tetracycline, macrolide-lincosamide-streptogramin B (MLSB), multidrug-resistant (MDR) and vancomycin-resistant genes in the aerosols. The most dominant drug class was beta-lactam and the genes were IMP-2-group (0.85), Per-2 group (0.65), OXA-54 (0.57), QnrS (0.50) and OXA-55 (0.55) in the urban non-clinical settings. The indoor aerosols possessed a richer diversity (Observed, Chao1, Shannon's and Pielou's evenness) of ARGs compared to the outdoors. Seasonal variations (autumn vs. winter) in relative abundances and types of ARGs were also recorded (R2 of 0.132 at p < 0.08). The presence of ARGs was found in both the inhalable (2.1 µm, 1.1 µm, 0.7 µm and < 0.3 µm) and respirable (>9.0 µm, 5.8 µm, 4.7 µm and 3.3 µm) size fractions within hospital aerosols. All the ARGs are of pathogenic bacterial origin and are hosted by pathogenic forms. The findings present baseline data and underpin the need for detailed investigations looking at aerosol as a vehicle for ARG dissemination among human and non-human terrestrial biota.

Assessment of mastitis in camel using high-throughput sequencing.

Camel milk is recognized as a functional food with significant economic value. Mastitis is one of the most common and costly diseases in the dairy industry. Mastitis, which is caused by pathogens such as bacteria, viruses, fungi, and algae, has an impact on the quality and quantity of milk produced as well as animal health and welfare. There is a paucity of data on the etiological factors that cause camel mastitis. This study reports the bacterial and fungal community involved in clinical camel mastitis using Illumina amplicon sequencing. A total of 25 milk samples were analyzed, including 9 samples with mastitis and 16 healthy samples. The bacterial community in healthy samples was significantly more diverse and abundant than in mastitis samples. The fungal population in mastitis samples, on the other hand, was more diverse and abundant. As compared to healthy samples, the genera Staphylococcus, Streptococcus, Schlegelella, unclassified Enterobacteriaceae, Lactococcus, Jeotgalicoccus. and Klebsiella were found to be abundant in mastitic milk. However, the genera Corynebacterium, Enteractinococcus, unclassified Sphingomonadaceae, Atopostipes, Paenibacillus, Pseudomonas, Lactobacillus, Sphingomonas, Pediococcus and Moraxella were reduced. In the fungal community, mastitis caused a significant increase in the relative abundance of the majority of taxa, including Candida, Phanerochaete, Aspergillus, Cladosporium and unclassified Pyronemataceae, while Penicillium and Alternaria showed a decline in relative abundance. In the bacterial and fungal communities, the discriminant analysis showed 19 and 5 differently abundant genera in healthy milk and mastitic milk, respectively. In conclusion, this study showed a microbiome dysbiosis linked to clinical camel mastitis, with opportunistic pathogens outgrowing commensal bacteria that were reduced. These findings are essential in designing an appropriate control program in the camel dairy herd, as well as in preventing and treating camel mastitis.

Data on microbial diversity of camel milk microbiota determined by 16S rRNA gene sequencing.

Raw camel milk samples were collected from three geographical locations (south, north and middle Kuwait) during two seasons. Next generation sequencing of the V3-V4 regions of the 16S rRNA gene was used to analyze the bacterial community in camel milk. DNA was extracted from one hundred thirty-three samples, and libraries were prepared using custom fusion primers of the 16S rRNA gene and sequenced on Illumina HiSeq 2500 platform. 16S rRNA gene sequences were aligned against the SILVA database SSU release 138. The high-throughput sequencing data are available at the NCBI database under the Bioproject PRJNA814013. This work describes camel milk's bacterial diversity among different geographical locations and seasons. The distribution of alpha diversity measures among camel milk sample groups collected from different geographical locations and seasons is presented. A significant effect of these parameters on camel milk's bacterial diversity was shown. Linear discriminant analysis (LefSe) showed significant differentially abundant bacteria at the phylum, class, order, family and genus level among the three locations and seasons. LefSe identified a total of 83 and 40 differentially abundant genera in the different geographical locations and seasons, respectively. More details about the bacterial composition of raw camel milk at the phylum and genus level can be found in research article [1]. These data can be used to compare the diversity of milk bacterial community between different milk producing species and camels from different parts of the world. Besides, these findings will contribute to our understanding of the camel microbiome structure and might be useful for designing an appropriate control program in the camel dairy herd. The data described in this article are available in Mendeley Data [2].

Synthesis, and characterization of metallic glassy Cu-Zr-Ni powders decorated with big cube Zr2Ni nanoparticles for potential antibiofilm coating applications.

Biofilms, are significant component that contributes to the development of chronic infections, especially when medical devices are involved. This issue offers a huge challenge for the medical community since standard antibiotics are only capable of eradicating biofilms to a very limited degree. The prevention of biofilm formation have led to the development of a variety of coating methods and new materials. These methods are intended to coat surfaces in such a way as to inhibit the formation of biofilm. Metallic glassy alloys, in particular, alloys that include copper and titanium metals have gained popularity as desirable antibacterial coating. Meanwhile, there has been a rise in the use of the cold spray coating technique due to the fact that it is a proper approach for processing temperature-sensitive materials. The present study was carried out in part with the intention of developing a new antibiofilm metallic glassy consisting of ternary Cu-Zr-Ni using mechanical alloying technique. The spherical powders that comprised the end-product were utilized as feedstock materials for cold spray coatings to stainless steel surfaces at low temperature. When compared to stainless steel, substrates coated with metallic glassy were able to significantly reduce the formation of biofilm by at least one log.

Camel milk microbiota: A culture-independent assessment.

Camel milk is renowned for its nutritional value and its therapeutic properties. It is considered a promising alternative to bovine milk due to its higher nutritional benefits, hypoallergenic characteristics and greater digestibility in the human gastrointestinal system. This study reports camel milk's bacterial and fungal microbiota, and the effect of geographical location and season on its bacterial community. We sequenced the V3-V4 regions of the16S rRNA gene for bacteria and the internal transcribed spacer (ITS) for fungi. A total of 134 samples of dromedary raw camel milk were collected from south, north and middle Kuwait during two seasons. Raw camel milk showed a diversified bacterial community, with 1196 genera belonging to 33 phyla. The four most predominant phyla of bacteria were Proteobacteria, Firmicutes, Actinobacteria and Bacteroidota. The core microbiota of raw camel milk, represented by the dominant genera shared by the majority of samples, was constituted by the genera Schlegelella, Paenibacillus, Lactobacillus, unclassified Comamonadaceae, Pediococcus, Moraxella, Acinetobacter, Staphylococcus, Enterococcus, Pseudomonas, Streptococcus, unclassified Micrococcaceae, Rothia, unclassified Sphingomonadaceae, unclassified Neisseriaceae and Sphingomonas. The fungal population was assessed in 14 raw camel milk samples, and comprised 87 genera belonging to 3 phyla. The genera Penicillium, Cladosporium, Candida, Aspergillus, Alternaria and Fusarium, dominated the fungal community. These findings shed light on raw camel milk's core bacterial and fungal microbiome. The geographical location and the season had a significant impact on the diversity and composition of camel milk microbiome.

Mechanical Alloying Integrated with Cold Spray Coating for Fabrication Cu50(Ti50-xNix), x; 10, 20, 30, and 40 at.% Antibiofilm Metallic Glass Coated/SUS304 Sheets.

Antibacterial agents derived from conventional organic compounds have traditionally been employed as a biofilm protective coating for many years. These agents, on the other hand, often include toxic components that are potentially hazardous to humans. Multiple approaches have been investigated over the last two decades, including the use of various metallic and oxide materials, in order to produce a diverse variety of usable coating layers. When it comes to material coating approaches, the cold spray technique, which is a solid-state method that works well with nanopowders, has shown superior performance. Its capacity to produce unique material coating in ways that are not possible with other thermal methods is the primary reason for its importance in contemporary production. The present work has been addressed in part to explore the possibility of employing mechanically alloyed Cu50(Ti50-xNix)x; x = 10, 20, 30, and 40 at.% metallic glass powders, for producing an antibiofilm/SUS304 surface protective coating, using the cold spray approach. In this study, elemental Cu, Ti, and Ni powders were low-energy ball milled for 100 h to fabricate metallic glassy powders with different Ni contents. The as-prepared metallic glassy powders were utilized to coat SUS304 sheets, using the cold spraying process. With high nanohardness values, the as-fabricated coating material, in particular Cu50Ti20Ni30, demonstrated remarkable performance in comparison to other materials in its class. Furthermore, it displayed excellent wear resistance while maintaining a low coefficient of friction, with values ranging from 0.32 to 0.45 in the tested range. E. coli biofilms were formed on 20 mm2 SUS304 sheet coated coupons, which had been injected with 1.5 108 CFU mL-1 of the bacterium. With the use of nanocrystalline Cu-based powders, it is feasible to achieve considerable biofilm inhibition, which is a practical strategy for accomplishing the suppression of biofilm formation.

Insights into Bacterial Community Involved in Bioremediation of Aged Oil-Contaminated Soil in Arid Environment.

Soil contamination by hydrocarbons due to oil spills has become a global concern and it has more implications in oil producing regions. Biostimulation is considered as one of the promising remediation techniques that can be adopted to enhance the rate of degradation of crude oil. The soil microbial consortia play a critical role in governing the biodegradation of total petroleum hydrocarbons (TPHs), in particular polycyclic aromatic hydrocarbons (PAHs). In this study, the degradation pattern of TPHs and PAHs of Kuwait soil biopiles was measured at three-month intervals. Then, the microbial consortium associated with oil degradation at each interval was revealed through 16S rRNA based next generation sequencing. Rapid degradation of TPHs and most of the PAHs was noticed at the first 3 months of biostimulation with a degradation rate of pyrene significantly higher compared to other PAHs counterparts. The taxonomic profiling of individual stages of remediation revealed that, biostimulation of the investigated soil favored the growth of Proteobacteria, Alphaprotobacteria, Chloroflexi, Chlorobi, and Acidobacteria groups. These findings provide a key step towards the restoration of oil-contaminated lands in the arid environment.

Integrated Phenotypic-Genotypic Analysis of Candidate Probiotic Weissella Cibaria Strains Isolated from Dairy Cows in Kuwait.

Probiotics represent a possible strategy for controlling intestinal infections in livestock. Members of the Weissella genus are increasingly being studied for health-related applications in animals and humans. Here we investigated the functional properties of two Weissella cibaria strains isolated from cows reared in Kuwait breeding facilities by combining phenotypic with genomic analyses. W. cibaria SP7 and SP19 exhibited good growth in vitro under acidic conditions and in the presence of bile salts compared to the reference probiotic Lacticaseibacillus (formerly Lactobacillus) rhamnosus GG. Both strains were able to adhere to Caco-2 and HT-29 cell lines, as well as to mucin. The cell-free supernatants of the two isolates exhibited inhibitory activity towards Escherichia coli ATCC 25,922 and Salmonella enterica UC3605, which was ultimately due to the low pH of supernatants. W. cibaria SP19 showed a co-aggregation ability similar to that of L. rhamnosus GG when incubated with S. enterica. Whole genome sequencing and analysis revealed that both strains harbored several genes involved in carbohydrate metabolism and general stress responses, indicating bacterial adaptation to the gastrointestinal environment. We also detected genes involved in the adhesion to host epithelial cells or extracellular matrix. No evidence of acquired antibiotic resistance or hemolytic activity was found in either strain. These findings shed light on the potential of W. cibaria for probiotic use in livestock and on the mechanisms underlying host-microbe interaction in the gut. W. cibaria` strain SP19 exhibited the best combination of in vitro probiotic properties and genetic markers, and is a promising candidate for further investigation.

Characterization and application of antimicrobials produced by Enterococcus faecium S6 isolated from raw camel milk.

The emergence of antimicrobial resistance in the food chain and the consumer's demand for safe food without chemical preservatives have generated much interest in natural antimicrobials. Thus, our main goal was to study the mode of action of the crude extract, the enterocins, and the organic acid produced by a bacteriocinogenic Enterococcus faecium strain S6 previously isolated from raw camel milk. Then, we aimed to evaluate their potential application in a food system. These antimicrobials exhibited antimicrobial activity against Listeria monocytogenes, Salmonella enterica, and Escherichia coli. The enterocins were synthesized as primary metabolites beginning at the lag phase, with optimal production at the exponential and stationary phases. The antimicrobials had a direct effect in extending the lag phase of L. monocytogenes, along with a significant inhibitory activity. The organic acid, in particular, inhibited both L. monocytogenes and S. enterica by inducing a total lysis and damage of the cell wall. The enterocins acted on disrupting the cell wall with pore formation, leading to cell death. Moreover, the crude extract revealed a combined inhibitory activity between enterocins and organic acid. Furthermore, the antimicrobials showed promising results through inhibiting L. monocytogenes cells in milk samples up to 1 wk at 4°C.

Characterization of semipurified enterocins produced by Enterococcus faecium strains isolated from raw camel milk.

Food safety has become an issue of great interest worldwide. Listeria monocytogenes is a food-borne pathogen that causes listeriosis and is difficult to control in the dairy industry. The use of lactic acid bacteria (LAB) and their antimicrobial substances against Listeria is promising in food applications. Here, we report the isolation from raw camel milk of LAB displaying antilisterial activity. Two isolates were selected for their secretion of bacteriocin(s) and identified by 16S rRNA sequencing as Enterococcus faecium S6 and R9. The produced bacteriocins were partially purified by ammonium sulfate precipitation and then biochemically characterized. Antimicrobial activity was estimated to be 6,400 and 400 AU (arbitrary units)/mL for E. faecium S6 and R9, respectively. The proteinaceous nature of the bacteriocins was confirmed via enzymatic reactions. Moreover, lipolytic and glycolytic enzymes completely inactivated the antimicrobial effect of the bacteriocins. These bacteriocins were heat-resistant and stable over a wide range of pH (2.0 to 10.0). To confirm its inactivation by lipolytic and glycolytic enzymes, the bacteriocin of E. faecium S6 was further purified by gel filtration, which suggested the existence of carbohydrate and lipid moieties. In addition, enterocin-coding genes were identified by PCR, showing DNA fragments corresponding in size to enterocins A, B, and P for E. faecium S6 and to enterocins B and P for E. faecium R9. In conclusion, these results indicate that partially purified bacteriocins from E. faecium S6 and R9 may be beneficial in controlling Listeria in the dairy industry.