Search Results (114)

Background/Objectives: Zoonotic tuberculosis (TB) remains a persistent public health challenge worldwide. It is particularly common in settings with close human–livestock–environment interactions. In Tanzania, progress toward TB control is increasingly threatened by multidrug-resistant tuberculosis (MDR-TB), yet genomic data from regions characterized by pastoralist and mining activities remain scarce. This study employed whole-genome sequencing (WGS) to characterize M. tuberculosis complex (MTBC) strains circulating among human and cattle populations in the Manyara region of northern Tanzania, with a focus on resistance-associated mutations and phylogenetic relationships. Methods: This cross-sectional study was conducted between September 2024 and February 2025. A total of 178 presumptive human TB cases provided sputum samples. From cattle, 161 samples were collected (110 milk samples and 51 lymph node aspirates), with each animal contributing only one type of sample. Specimen were analyzed using GeneXpert MTB/RIF, Lowenstein–Jensen culture, and WGS. Phylogenetic reconstruction was performed using SNP-based methods and IQ-TREE2 version 2.2.0. Results: Among human participants, 14 (7.8%) sputum samples were GeneXpert positive and were confirmed as members of MTBC by LJ culture. In cattle, one (0.62%) lymph node aspirate was positive for MTBC. Significant predictors of MTBC positivity included previous TB history, weight loss, and occupation involving mining and cattle keeping. WGS of five human isolates identified Lineages 1, 3, and 4. One isolate (Sample 98) harbored mutations associated with XDR-TB. Conclusions: WGS revealed M. tuberculosis Lineages 1, 3 and 4 circulating in the Manyara region, with diverse genetic mutations conferring with resistance to first- and second-line anti-TB drugs. These findings highlight the importance of integrated genomic surveillance to monitor drug resistance patterns in Tanzania and similar settings across human and animal populations. Full article

Background: Bovine milk protein-based human milk fortifiers (HMFs) are routinely used in preterm infants in low- and middle-income countries (LMICs) but are associated with potential adverse effects. Plant-derived amino acid-based HMFs are a promising alternative, but comparative data from randomized controlled trials (RCTs) are lacking. This pilot RCT aimed to evaluate the feasibility, safety, and growth outcomes of a novel plant-derived amino acid-based HMF compared to a bovine-derived whole-protein-based HMF in preterm infants at <34 weeks’ gestation. Methods: In this open-label, parallel pilot RCT, preterm infants at <34 weeks’ gestation were randomized to a plant-derived amino acid-based HMF (n = 66) or a bovine-derived whole-protein-based HMF (n = 70). Primary outcomes were time to reach 180 mL/kg/d enteral feeds and growth velocities to discharge and 40 weeks’ postmenstrual age (PMA). Feasibility and safety were also assessed. The analysis followed an intention-to-treat approach. The trial was registered (CTRI/2025/06/089133). Results: Baseline characteristics were comparable between groups. Times to reach enteral feeds of 180 mL/kg/d did not differ (adjusted Hazard Ratio 0.93, 95% CI 0.65, 1.33; p = 0.70). In-hospital weight gain was significantly higher in the plant-derived HMF group (adjusted mean difference (aMD) 3.20 g/kg/d, 95% CI 0.46, 5.95; p = 0.02). This benefit was most evident in subgroup analyses of infants with birth weight ≥1500 g (aMD 4.47 g/kg/d, 95% CI 0.09, 8.84; p = 0.04). Length, head circumference growth, and all safety outcomes were similar between groups. Conclusions: The plant-derived amino acid-based HMF demonstrated clinical feasibility and was well-tolerated. While it was associated with better in-hospital weight gain, differences in overall macronutrient density between the formulations may account for this finding. These results are hypothesis-generating and warrant future adequately powered trials. Full article

Gastric cancer (GC) remains a major global health challenge, ranking among the most lethal malignancies due to late diagnosis, high tumor heterogeneity, and limited treatment efficacy. The search for safer, nutritionally based adjuncts to conventional therapies is therefore a research priority. Donkey milk (DM), traditionally used as a hypoallergenic substitute for infants, is emerging as a functional food with remarkable bioactivity. Its composition closely resembles human milk, with high levels of bioactive proteins, a favorable polyunsaturated lipid profile, antioxidant vitamins, and immune-supportive minerals. Despite its growing nutraceutical appeal, the anticancer potential of DM in GC has not yet been explored. This study represents the first investigation of DM in human gastric adenocarcinoma (AGS) cells. Using increasing concentrations of whole DM (25–100%), a dose-dependent inhibition of cell viability and migration was observed. Mechanistic insights reveal that DM induces mitochondrial oxidative stress, disrupts cell cycle progression (S/G2 accumulation at 75%, G2 arrest at 100%), and unexpectedly triggers a pro-inflammatory gene signature suggesting stress-driven immunostimulation rather than canonical apoptosis. These findings highlight a non-classical, context-dependent cytotoxic mechanism that distinguishes DM from conventional pro-apoptotic agents. DM may represent a promising nutraceutical candidate for GC management, bridging traditional food resources with modern oncology. By inhibiting hallmark cancer traits while engaging unique immunological pathways, DM offers a sustainable, low-toxicity approach with translational potential. Future studies will focus on the characterization of active components, validation in organoid and animal models, and exploring clinical applications of DM-derived bioactive components in cancer prevention and therapy. Full article

Background/Objectives: Streptococcus agalactiae is one of the most significant pathogens causing infections in humans and mastitis in dairy cattle. This work focused on a comprehensive comparative pan-genomic analysis of bovine and human Group B Streptococcus to elucidate the genetic mechanisms underlying host adaptation and dissemination. Methods: Isolates of S. agalactiae from quarter milk samples from dairy herds in Lombardy (Italy), along with human strains, were considered. Whole genome sequencing was used to compare core and accessory genomes, assign sequence types, and find virulence and resistance factors. Results: 30 sequence types were detected, of which two (ST12 and ST23) in common between bovine and human. The allele frequencies for resistance determinants revealed elevated rates for tetM (59.8% overall, 66.5% in human clinical isolates), ermB (17.3% overall, 20.4% in human clinical isolates), and ant(6)la (8.9% overall, 12.8% in human clinical isolates). Bovine strains had accessory gene clusters linked to lactose metabolism and immunological evasion, whereas human isolates were concentrated in regions related to adhesion and antibiotic resistance. Conclusions: Comparative pan-genomics show that there is a small genetic overlap between bovine and human Group B Streptococcus populations. Full article

The Enterobacteriaceae family is a significant source of foodborne pathogens and represents a severe threat to human and animal health. These bacteria can penetrate the dairy supply chain through direct contact with cattle and the livestock environment and can survive production processes. Escherichia coli (E. coli), one of the most diffuse bacteria in raw and processed milk, exposes consumers to the risk of contaminated milk. As a result of this exposition, several milk-borne illness outbreaks have been reported worldwide, underscoring the urgent need for effective detection and prevention measures. Conventional analysis methods are effective but have significant limitations, including the requirement of pre-treatment and pre-enrichment steps. Thus, the need for advanced detection techniques that can accurately identify these pathogens without pre-treatment steps is critical. In this work, a proof-of-concept biosensor based on a spoon-shaped optical biochip was developed to detect E. coli via surface plasmon resonance (SPR) phenomena and was combined with a polyclonal antibody layer against E. coli as a molecular recognition element (MRE). The proposed label-free biosensing strategy, achieved by exploiting simple SPR spoon-shaped biochips, exhibits a remarkable detection limit (6.8 colony-forming units, CFU/mL) and high specificity towards other interfering bacteria belonging to the Enterobacteriaceae family. In addition, tests on commercial milk samples were carried out, achieving recovery values of 95% and 102% for whole milk and infant milk, respectively. The proposed spoon-shaped biosensor enables label-free biosensing without the need for microfluidic systems. It provides a rapid response (10 min), paving the way for its use as a point-of-care test (POCT) in real-world settings. Full article

Introduction: Breast milk is the best food for a growing infant during the first 6 months of life. The presence of trans fatty acids (TFAs) in breast milk can interfere with the synthesis of long-chain polyunsaturated fatty acids (LC-PUFAs), increasing the risk of developing, among other issues, asthma or atopic dermatitis. TFAs are not synthesised de novo in the human body. Their content in breast milk may be a good marker of short-term dietary intake of these compounds by breastfeeding mothers. However, the literature shows differences in the assessment of the relationship between dietary TFAs intake and TFAs content in breast milk. Furthermore, the decrease in the TFAs content in food observed recently seems to make it impossible to use Craig-Schmidt’s formula to estimate the TFAs content in the diet or in breast milk. The aim of this study was to confirm the possibility of using TFAs content in breast milk as a marker of their short-term intake within the breastfeeding mother’s diet, together with an attempt at preliminary quantitative determination of the relationship between these parameters. Materials: The study material was collected from a single breastfeeding mother and included 10 breast milk samples and 10 samples of daily food rations reconstructed based on the 24 h food consumption survey. Methods: The content of fatty acids, including TFAs, was determined by gas chromatography–mass spectrometry (GC-MS). Results: The TFAs content in the whole-day mother’s diet and in 100 mL of breast milk ranged from 0.11 to 0.54 g/day and from 0.02 to 0.07 g, respectively. A strong statistically significant (p < 0.05) positive correlation between these parameters was found. Equations for an exploratory linear relationship between the TFAs content in a breastfeeding mother’s diet and the concentration of these fatty acids in breast milk have been developed. Due to the small number of samples, these data should be interpreted very cautiously and validated in a larger cohort. Conclusions: This single-participant pilot study suggests that TFAs content in breast milk may be a marker of the dietary intake of these compounds from the previous day. It seems that the dietary habits of breastfeeding mothers of twins are a significant factor influencing the composition of breast milk and, consequently, the nutritional quality of breastfed infants. Full article

Camel milk is widely consumed in the world’s arid and semi-arid regions because of its favorable nutritional profile and associated human health benefits. The indigenous microbiota of raw camel milk is diverse and composed of different bacterial and fungal groups. This community drives spontaneous milk fermentation, resulting in a variety of traditional products, including Gariss, Shubat, Chal, Dhanaan, Lfrik, and Suusac (or Suusa), depending on geographic region and cultural practice. This fermented milk has improved sensory, nutritional, and health profiles, as well as an extended shelf life, compared to raw milk. Fermentation alters the microbial community structure, with lactic acid bacteria (LAB) consistently becoming dominant, while yeasts and molds are also detected in some products. These patterns have been identified using both culture-dependent and culture-independent approaches, including 16S rRNA gene sequencing and whole-genome shotgun metagenomics. However, the milk’s microbial composition is highly variable and is influenced by the original composition, geographical location, fermentation and hygiene practices. The detection of opportunistic pathogens such as E. coli, Salmonella and Listeria in some traditional products raises important food safety concerns. This review presents current knowledge on fermented camel milk microbiology using a cross-regional approach, identifying key gaps in microbial safety and process standardization to support wider acceptance and potential commercialization. Full article

To date, the problem of mastitis in cattle remains relevant for both the industrial sector and scientific research. Despite numerous active investigations, the causes of this disease have not been fully established. It is postulated that several factors may be involved, such as bacterial pathogens, animal husbandry practices, and weather and climatic conditions. In this study, we selected cows from farms in Yakutia to investigate microbial isolates present in the milk of cows affected by mastitis and treated with antibiotics. Five identified Staphylococcus aureus isolates were investigated using whole-genome sequencing (Illumina sequencing and nanopore sequencing), followed by analysis of virulence factors in the genomes and cultural properties of the isolates. The profile of S. aureus virulence genes (exotoxins, cytotoxins, superantigen-like proteins, adhesins) was identified via WGS. Hemolysin gene (hla) was detected in all isolates. An investigation of the cultural properties of the isolates, specifically through hemolysis of rabbit erythrocytes and Western blot analysis of the culture liquid of S. aureus, revealed different expression levels of alpha-hemolysin among the strains. One isolate (17-21) exhibited the highest secretion level of about 320 ± 37 ng, both in the hemolysis test and immunoblotting assay. An investigation of the isolates’ antibiotic resistance showed that all isolates exhibited multidrug resistance, as confirmed by the presence of antibiotic resistance genes in these isolates. One isolate (7-7) exhibited the broadest range of phenotypic resistance and was resistant to all tested antibiotics (except clindamycin). Phylogenetic analysis suggested that the evolution of these isolates occurred independently in their respective ecological niches, although their transfer from cattle to humans, and vice versa, is possible. Isolates 7-7, 18-22, 33-40, and 35-42 are most typical to Yakutian cattle, while isolate 17-21 might have been introduced from a different region. To the best of our knowledge, this is the first in-depth study into a range of S. aureus isolates associated with mastitis infection in Yakutian cattle. Full article

Background/Objectives: Appetite-regulating hormones are bioactive components of human milk. We tested the associations of leptin and adiponectin with infant growth and eating behaviors to age 6 months. Methods: In a cohort of 70 healthy, full-term infants and their mothers, human milk adiponectin and leptin were assayed at age 2 months (m). At infant ages 2, 4, and 6 m, infant anthropometry was obtained, mothers reported feeding frequency, duration, and breastfeeding intensity and completed the Baby Eating Behavior Questionnaire (Enjoyment of Food, Food Responsiveness, and General Appetite), and infant sucking vigor using an artificial nipple (burst duration and sucking frequency) was measured. Mothers reported demographics, gestational diabetes and pre-pregnancy body mass index (BMI), gestational age, and infant birthweight. Multivariate models evaluated predictors of leptin and adiponectin, and associations of leptin and adiponectin with infant growth and eating behaviors. Results: Human milk leptin was predicted by maternal BMI (β = 0.02) and breastfeeding intensity (β = −0.32). Regarding infant growth, infant weight-for-age and weight-for-length z-scores at 6 m were predicted by leptin (β = 0.91 and β = 1.22, respectively) and adiponectin (β = 0.01 and β = 0.01, respectively). Regarding infant eating behaviors, feeding duration at 2 m and feeding frequency at 4 m were predicted by adiponectin (β = 0.03 and β = −0.02, respectively). Conclusions: Human milk leptin and adiponectin may contribute to weight gain in early infancy, but the effect does not appear to be mediated substantially by infant eating behaviors. Further investigation into the metabolic programming of early infant weight gain is warranted. Full article

Background/Objectives: Antimicrobial agents have revolutionized disease management in humans and animals; however, their misuse and overuse have accelerated the emergence and spread of antimicrobial resistance (AMR) and antimicrobial resistance genes (ARGs). Dairy farms are recognized as potential hotspots for ARG dissemination, particularly through Escherichia coli, which acts as a reservoir and vector of ARGs, enabling their horizontal transfer via plasmids and other mobile genetic elements. This study aimed to characterize the genomic diversity, ARG profiles, plasmid content, and phylogenetic relationships of E. coli isolated from dairy farm environments and milk using whole-genome sequencing. Methods: A total of 31 E. coli isolates recovered from soil, effluent, cow dung, and milk samples underwent deoxyribonucleic acid extraction, library preparation, and sequencing on the Illumina MiSeq platform, followed by comprehensive bioinformatic analysis. Results: The E. coli isolates exhibited 20 distinct sequence types, including one novel sequence type. Plasmids were detected in 71% of the isolates, with the IncF plasmid family being the most predominant. Furthermore, 12 ARG groups were identified, with β-lactam resistance genes detected in 67.7% of isolates. Notably, blaCTX-M genes were identified in all phenotypically confirmed extended-spectrum β-lactamase-producing isolates. Additional ARGs, including those conferring resistance to tetracyclines (tet(A), tetX4), quinolones (qnrS1), aminoglycosides (aph, aad, ant), and folate pathway inhibitors (dfr and sul), were widely distributed throughout the samples. Phylogenetic analysis revealed clustering of isolates from different sample types, particularly among ST58 isolates, suggesting cross-environmental transmission. Conclusions: This study demonstrates that E. coli from dairy farm environments harbor diverse ARGs and plasmids, confirming their role as reservoirs of AMR. These findings underscore the importance of prudent antimicrobial use, routine genomic surveillance, and enhanced biosecurity measures to limit cross-environmental transmission. Full article

Background/Objectives: Escherichia coli is a ubiquitous commensal organism in humans, animals, and the environment, but certain strains harbour virulence and antimicrobial resistance (AMR) determinants that can cause significant disease. Food-producing animals, including dairy cattle, may act as reservoirs for AMR E. coli, and raw milk and raw milk products can serve as potential exposure pathways to humans. However, data on the prevalence and genomic characteristics of AMR E. coli in raw milk in Ireland are limited. This study aimed to describe the occurrence of commensal and clinically relevant AMR E. coli in raw milk and raw milk dairy products in Ireland and to characterise their antimicrobial susceptibility and genetic characteristics. Methods: A total of 139 raw milk and raw milk dairy product samples were collected and analysed for commensal E. coli and fluoroquinolone-resistant, extended-spectrum β-lactamase (ESBL)/AmpC β-lactamase and carbapenemase-producing E. coli. AMR patterns were determined in line with EU surveillance guidelines based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines which use minimum inhibitory concentration (MIC) breakpoints. Whole genome sequencing (WGS) was conducted on selected isolates to identify AMR genes (ARG), virulence factors, plasmid replicons, efflux pump, disinfectant resistance genes, multi-locus sequence types (MLSTs) and phylogenetic diversity. Results: A total of forty-seven E. coli isolates were recovered (33.8% isolation rate). Thirteen isolates exhibited resistance to between two and nine antimicrobials, with twelve classified as multidrug resistant (MDR). The highest resistance frequencies were to ampicillin, sulfamethoxazole, trimethoprim and tetracycline. Four fluoroquinolone-resistant isolates, one ESBL producer (bla_CTX-M-3), and one carrying a AmpC promoter mutation were identified; no carbapenemase producers were detected. WGS revealed diverse sequence types, multiple virulence determinants, plasmid replicons, intrinsic efflux pump genes, and limited presence of the disinfectant resistance gene qacEΔ1. Conclusions: Raw milk and raw milk dairy products in Ireland can harbour AMR E. coli, including MDR and potentially pathogenic strains, highlighting the need for ongoing surveillance within the dairy supply chain. Full article

Background/Objectives: Staphylococcus aureus is a multifaceted pathogen responsible for diseases in humans and in several animal species, including dairy cows. This study aimed to characterize and compare the genetic diversity, lineage distribution, and antimicrobial resistance profiles of S. aureus isolates from bovine milk with human-derived reference genomes to investigate host adaptation and inter-species transmission. Methods: Genomic analyses were performed on S. aureus isolates from quarter milk samples of dairy cows together with human-derived sequences from public databases. Whole-genome sequencing and multi-locus sequence typing (MLST) were used to determine sequence type (ST) distribution, and the presence of key antibiotic resistance genes and mobile genetic elements (MGEs) was assessed. Comparative genomics was applied to evaluate gene content, phylogenetic relationships, and lineage–host associations. Results: The dataset encompassed bovine-adapted lineages (CC97, CC133, CC151) and human-associated lineages (CC1, CC5, CC8, CC30, CC45), as well as livestock-associated ST398 in bovine samples and human-adapted ST5 and ST6 in animals. ST8 was the only ST shared between animal and human isolates and showed differing resistance profiles, with animal ST8 carrying resistance determinants absent from human ST8. Bovine-adapted strains were characterized by recurrent loss of human-associated virulence genes and acquisition of bovine-associated mobile genetic elements, and blaZ and mecA were rarely detected in bovine-adapted CC97 but were frequently present in human CC5 and CC8. Overall, animal isolates carried fewer resistance genes than human isolates. Conclusions: S. aureus from dairy cows and humans displayed substantial genetic diversity, with evidence of host-associated lineages and dynamic changes in gene and mobile element content. These findings support the need for integrated One Health surveillance to track shared and host-adapted lineages and their antibiotic resistance determinants. Full article

3′-sialyl-N-acetyllactosamine (3′-SLN) and 6′-sialyl-N-acetyllactosamine (6′-SLN) are two important human milk oligosaccharides (HMOs) which play significant functions in brain development and antiviral potential. However, their metabolism is still unknown. In this study, chemoenzymatically synthesized 3′-SLN and 6′-SLN were labeled with cyanine-7 (Cy7) via formation triazole (Tz) derivatives to investigate their metabolism and organ distribution in a mouse model. The fluorescence signals were detected in the brains of mice after 0.5 h of gavage with 3′-SLN-Tz-Cy7 and 6′-SLN-Tz-Cy7. It was found for the first time that both of them can cross the blood–brain barrier (BBB) as a whole and reach the brain in a sex-specific manner. And the results show that, whether in the in vivo imaging results or the brain fluorescence signal results, the male mice absorbed 3′-SLN-Tz-Cy7 more than 6′-SLN-Tz-Cy7; meanwhile, in the female mice, the results were exactly the opposite. Both 3′-SLN-Tz-Cy7 and 6′-SLN-Tz-Cy7 exhibited the highest fluorescence intensity in pulmonary tissues, followed by substantial hepatic deposition. This study would offer preliminary evidence for the hypothesis that the oral administration of 3′-SLN and 6′-SLN may promote brain development and provide a foundation for the further exploration of their functions in brain cognition. Full article

Background/Objectives: Streptococcus uberis is a Gram-positive bacterium and a major cause of bovine mastitis. The use of antimicrobial treatments raises concerns about resistance. This study aimed to characterize S. uberis isolates from one of the ten largest milk-producing regions in Europe. Methods: Thirty-six isolates from 36 cows with mastitis were identified using MALDI-TOF and VITEK^®MS. Susceptibility to 9 antibiotics (penicillin G, ampicillin, tetracycline, erythromycin, clindamycin, cefotaxime, ceftriaxone, levofloxacin, and moxifloxacin) was determined with VITEK^®2. Whole-genome sequencing was performed using MinION Mk1C. Results: Alleles were identified for 7 loci: arcC, ddl, gki, recP, tdk, tpi, and yqiL. Only 10 isolates had alleles for all the loci. The loci with the highest number of alleles were ddl and tdk (33/36 strains), while arcC had the fewest (19/36). Four isolates were assigned to known sequence types (ST6, ST307, and ST184), and novel alleles were detected in 32 of the 36 isolates. Twelve isolates showed phenotypic resistance to one or more of the following antibiotics: tetracycline, erythromycin, clindamycin, and ceftriaxone. The lnu was the most frequently detected resistance gene (27 out of 102 total gene appearances). A total of 19 virulence factors were identified. All strains were predicted to be capable of infecting human hosts. Conclusions: Streptococcus uberis is a potential reservoir of antimicrobial resistance genes. The use of antimicrobials to treat bovine mastitis has reduced the susceptibility of this microorganism to several antibiotics, underscoring the importance of monitoring antimicrobial use in veterinary practice. The results also highlight the high genetic diversity of the isolates, suggesting a strong capacity to adapt to different environmental conditions. Full article

(1) Background: Water buffaloes (Bubalus bubalis) are important for dairy and meat production. Up to now, genomic analysis has focused on female subjects, leaving the Y chromosome essentially unknown. Advances in third-generation sequencing (TGS) made it possible to improve the study of complex genome sequences, such as buffalo and other mammalian species including humans. (2) Methods: In this study, we applied TGS-based long-read sequencing to generate, in one step, high-quality whole-genome sequences, which can take full advantage of a rapid bioinformatic pipeline, such as that described in the companion paper. (3) Results: Five male buffalo genomes have been fully sequenced at relatively high depth (20–40×) which, combined with the read length typical of TGS, provide the basis for important insights into male-specific genetic traits, including those linked to meat and milk production. (4) Conclusions: With the use of TGS technologies, we offer a complete strategy for fast, one-step genome sequencing which can also be applied to other farm animals with a comparably large genome. This approach can help in revealing genetic features characteristic of an animal individual beyond the simple assessment of a number of SNPs or other known sequence variations, thus supporting improved genetic selection for dairy productivity and future research on genetic variability in buffalo breeds. Full article