Search Results (911)

Accurate detection of Campylobacter in chicken liver is hindered by strong matrix inhibition. This study evaluated sample-processing strategies to improve droplet digital PCR (ddPCR) quantification of Campylobacter coli and Campylobacter jejuni in chicken liver. Mechanical homogenization (Stomacher) and enzymatic/mechanical dissociation (gentleMACS), with and without 8 μm filtration, were compared. Particle-size analysis showed that filtration, especially following gentleMACS treatment, produced smaller, more uniform particles and reduced variability. Percent-degradation assays confirmed that gentleMACS achieved substantially greater tissue disruption than Stomacher homogenization. The multiplex ddPCR assay, which simultaneously targets C. coli and C. jejuni, produced droplet counts comparable to single-target reactions, indicating minimal interference between targets under the conditions tested. In inoculated liver samples, gentleMACS processing yielded droplet counts similar to those obtained from pure cultures, whereas unprocessed liver caused severe matrix interference and inconsistent quantification. Furthermore, gentleMACS-treated samples exhibited strong log-to-log linearity for quantifying C. coli and C. jejuni, enabling detection near 1 genome copy equivalent per reaction. Overall, the results indicate that enzymatic/mechanical dissociation combined with fine-pore filtration improves ddPCR detection of Campylobacter species in chicken liver. Full article

Acute gastroenteritis is a common condition with a wide and often indistinguishable etiology, requiring laboratory support for an accurate diagnosis. Classical diagnostic methods, including stool culture and antigen-based tests, are limited by restricted pathogen coverage and variable sensitivity. In the present study, 548 stool samples from patients with suspected gastroenteritis were tested using the QIAstat-Dx Gastrointestinal Panel 2 and compared with stool culture and rotavirus/adenovirus antigen tests. The molecular panel showed a positivity rate of 50.4%, consistently higher than stool culture (12.6%) and antigen assays (12.3% for rotavirus and 4.4% for adenovirus). The most frequently detected pathogens included enteropathogenic Escherichia coli (15.3%), Campylobacter spp. (12.0%), and enteroaggregative E. coli (10.2%). Agreement between methods was good for bacterial pathogens but low for viral targets. Discordant viral results were often associated with low antigen cut-off index values and multiple pathogen detections by the molecular panel, suggesting potential limitations of antigen-based assays. Overall, the QIAstat-Dx Gastrointestinal Panel 2 improves pathogen detection and provides rapid, comprehensive diagnostic information, while a combined approach with conventional methods may represent the most appropriate strategy for optimizing patient management. Full article

Antimicrobial resistance (AMR) is one of the most serious threats to global public health, driven in part by extensive antibiotic use in food-producing animals. The poultry industry, a major contributor to the global animal protein supply, has depended on antibiotics for growth promotion and disease control, thereby contributing to the emergence and dissemination of AMR zoonotic bacteria. This review synthesizes current evidence on the potential of phytochemicals (PCs), plant-derived bioactive compounds, as sustainable non-antibiotic alternatives for controlling bacterial foodborne pathogens in poultry. Relevant literature including in vitro and in vivo studies assessing PCs against major poultry-associated zoonotic bacteria, including Salmonella enterica, Campylobacter spp., Clostridium perfringens, Listeria monocytogenes, and pathogenic Escherichia coli, is examined. Evidence indicates that PCs exert antimicrobial and anti-virulence effects through mechanisms like bacterial membrane disruption, inhibition of quorum sensing and virulence gene expression, modulation of gut microbiota, and enhancement of host immune responses. In vivo studies demonstrate reductions in pathogen colonization and improvements in gut health and performance metrics in poultry. Despite these promising findings, challenges remain in bioavailability, dose optimization, standardization, and regulatory approval. Overall, PCs represent a promising component of integrated antimicrobial stewardship strategies in poultry production, with significant implications for mitigating zoonotic AMR transmission. Full article

In recent years, diseases caused by species of the genus Campylobacter have increased, due to improvements in identification methods, but also because, as part of global travel and trade, these species have spread to countries where no cases had previously been reported. The methodologies for their identification, whether classic through culture media and morphological characteristics, or using molecular biology or even proteomics techniques, play a fundamental role in establishing their diagnosis and providing timely treatment. Likewise, epidemiology will help guide this diagnosis when dealing with poorly defined diarrhoea, as well as the control and prevention of these infections. Similarly, expanding information on the relationship between these species and Guillain-Barré syndrome will lead to a better understanding and timely identification. We must not forget that both intrinsic and acquired antimicrobial resistance are key factors to consider for the successful treatment of infections caused by Campylobacter species. Full article

Poultry is the main reservoir of Campylobacter spp. and most human cases result from consuming undercooked poultry or handling raw meat. In 2022, a total of 55 samples, including neck skin, cecal contents, and processing waters, were collected at two poultry slaughterhouses in Italy and analysed according to ISO 10272-2:2017 at the Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata laboratories. Overall, 51/55 (92.72%) samples tested positive for Campylobacter. Among the isolates, 64.71% were identified as C. coli, and 35.29% as C. jejuni. Phenotypic and genotypic analysis were performed to assess antimicrobial resistance and virulence characteristics. All C. jejuni isolates and 72.72% of C. coli showed resistance to fluoroquinolones. Resistances to tetracycline and carbapenem were observed in 60.78% and 45.09% of isolates, respectively. Genomic analysis confirmed the presence of the tet(O) gene, conferring tetracycline resistance. In addition, OXA-450 and OXA-466 genes, conferring beta-lactam resistance, were detected in 78.43% and 3.92% of isolates. Virulence-associated genes were detected. Specifically, the ciaB gene was found in 50/51 (98.04%) of isolates, whereas jlpA, cdtA, cdtB, and ctdC genes were exclusively identified in C. jejuni strains. The high prevalence of pathogenic and antimicrobial-resistant Campylobacter strains highlights the need for strengthened control measures along the poultry production chain. Full article

The double-crested cormorant (Nannopterum auritum), a piscivorous bird endemic to North America, frequently forages in aquaculture ponds during migration and wintering, contributing to economic losses in catfish-producing regions of the southern United States. While interactions between cormorants and aquaculture systems are well documented, their associated microbial communities and genetic elements remain less characterized. In this exploratory study, Hi-C-enabled metagenomics was applied to fecal samples from two cormorants to generate a genome-resolved, descriptive analysis of gut microbial composition and to associate bacterial genomes with mobile genetic elements (MGEs), antimicrobial resistance genes (ARGs), and putative virulence-associated genes. Metagenome-assembled genomes (MAGs) included taxa reported in aquatic or animal-associated environments, including Edwardsiella tarda, Plesiomonas shigelloides, Clostridium perfringens, and Campylobacter volucris. ARGs were detected across multiple MAGs, with E. tarda harboring the greatest diversity. Hi-C-enabled linkage of plasmids and phages to putative hosts, providing structural insight into microbial organization. Analyses are descriptive (n = 2) and do not include statistical comparisons or diversity metrics. These findings demonstrate the utility of Hi-C for resolving gene–host associations and provide a framework for future studies of microbial connectivity in One Health contexts. Full article

This study used three complementary datasets to investigate the relationship between human Campylobacter infections in Estonia and potential sources. A targeted dataset of 15 C. jejuni genomes with overlapping sequence types from human cases and broiler chicken meat was analysed using genotyping and in silico antimicrobial resistance profiling, alongside 20 human isolates for source attribution. Additionally, 12,111 isolates were analysed to provide population-level context. The core genome multilocus sequence typing showed a high similarity (less than three allelic differences) between the human and broiler isolates of ST122, ST464, and ST7355, indicating poultry as a likely source, whereas ST9882 was more divergent (13–18 allelic differences). The resistance profiles were consistent within ST122, ST464, and ST7355, and all were resistant to ciprofloxacin, nalidixic acid, ampicillin, and tetracycline, while ST9882 additionally exhibited aminoglycoside (streptomycin) resistance. The source attribution linked 77.8% of the human cases to chicken and 22.2% to cattle. A novel genotype, ST11001, was identified in humans and attributed to cattle source, while C. coli isolates were linked to birds and sheep. Poultry dominated the larger dataset (87.3%). Gastroenteritis was the predominant clinical presentation (98.5%), whereas ST22 and ST122 were associated with Guillain–Barré syndrome. These findings support poultry as a major reservoir of human Campylobacter infections and highlight the need for coordinated cross-border surveillance. Full article

The rise in global consumption of fresh vegetables is a response to their nutrient-dense composition and low caloric content—key factors for optimising human metabolic health. This study evaluated the Quantitative Microbial Risk Assessment (QMRA) of Helicobacter pylori and enteric pathogens in fresh vegetables within the central highlands of Peru. The research integrated conventional microbiology, qPCR, and Monte Carlo simulations. The results revealed a high prevalence of Escherichia coli (83.7%), with a heterogeneous distribution where Huancayo presented the highest prevalence (95.5%) and Chupaca the lowest (68.2%). In contrast, pathogens such as H. pylori and Campylobacter jejuni showed marginal prevalences of 2.33% and 3.49%, respectively, with detections restricted to leafy and root vegetables at specific points of sale. Although biochemical tests indicated the presumptive presence of Helicobacter pylori, the qPCR results were negative, possibly due to the bacteria’s viable but non-culturable (VBNC) state. The QMRA model showed a highly skewed annual infection risk distribution, with E. coli presenting the highest risk: median Pann = 1.000 and 84.3% of simulations exceeding the WHO tolerable threshold of 10⁻⁴. For Salmonella Typhimurium and Shigella flexneri, 22.4% and 9.1% of simulations exceeded the same threshold, respectively. The results underscore the urgent need to implement traceability programs and improve agricultural practices across the evaluated provinces. Full article

Meat serves as a prime medium for the growth of foodborne pathogens due to its rich protein content and high water activity, contributing significantly to the global burden of foodborne illnesses. This review synthesizes current advances in meat-borne bacterial pathogen detection with particular emphasis on emerging artificial intelligence (AI)-enabled applications. Major pathogens of concern, including Salmonella, Listeria monocytogenes, Escherichia coli, Campylobacter, Clostridium, and Staphylococcus aureus, are examined in relation to their relevance across the meat supply chain. Recent progress in biosensors (clustered regularly interspaced short palindromic repeats), CRISPR-based assays, isothermal amplification, and metagenomics is evaluated alongside the growing role of AI in automating signal interpretation, enhancing image-based diagnostics, and supporting early contamination prediction. AI-based systems have proved 96.4–104% recovery and 100% bacterial capture ability. Embedding AI methods in a wet lab demands technical and logical modeling, as well as learning and calibration decorum. Nonetheless, AI readiness and full-scale application for meat-borne pathogens surveillance are on the way. Furthermore, additional focus is aligned on meat-borne bacterial pathogen genomic databases, i.e., (NCBI Pathogen Detection, EnteroBase, VFDB, ComBase, and GenBank), which serve as critical training resources for AI models for outbreak tracking, virulence profiling, and antimicrobial resistance (AMR) prediction. By integrating molecular methods, genomic surveillance, and AI-driven analytics, this review presents a framework for strengthening meat safety systems. This will improve early detection capabilities and support data-driven public health interventions in the future. Full article

During Zambia’s 2023–2024 cholera outbreak, reliance on single-pathogen diagnostics risked overlooking co-circulating enteric pathogens. This study estimated the prevalence of rotavirus and described co-detected enteropathogens and rotavirus genotypes among patients admitted with suspected cholera. A sub-analysis was conducted on diarrhoeal stool specimens collected from patients who met the syndromic suspected cholera case definition. Samples were tested using the Bosphore^® Gastroenteritis Panel v2, a multiplex PCR enteric panel, to detect rotavirus and other gastrointestinal pathogens. Rotavirus-positive specimens with sufficient viral load were further genotyped by RT-PCR targeting of the VP7 and VP4 genes. Among 319 suspected cholera admissions, rotavirus was detected in 18 patients (5.6%; 95% CI 3.4–8.8%), predominantly in children aged <5 years (27.8%, 5/18) and 6–17 years (27.8%, 5/18). Co-infection was common, with 17/18 (94.4%) of rotavirus-positive samples showing co-infection with at least one additional enteric pathogen, most frequently Campylobacter. Genotyping was successful in five samples and revealed heterogenous circulating strains, including G1P[8], G2P[4], G3P[6], G12P[6], and G1P[6]. Rotavirus accounted for a modest proportion of suspected cholera admissions and was frequently detected in mixed enteric infections, underscoring the value of multi-pathogen diagnostics and continued molecular surveillance during outbreak response. Full article

Silage is a fundamental component of cattle feed, and its microbiological quality is critical for animal health and human safety. Improper ensiling conditions, such as oxygen exposure or inadequate acidification, can promote the growth of pathogens like Listeria monocytogenes, Clostridium botulinum, and Bacillus cereus. This study aimed to evaluate the microbial status of maize silages and identify pre-ensiling factors influencing its hygienic safety. Over a four-year period, 406 silage samples were collected from cattle farms across Poland. The research evaluated general hygiene indicators and screened for specific pathogens using standard culture methods, polymerase chain reaction toxotyping, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The impact of agricultural practices, including soil quality, organic fertilization, and microbial inoculation, was also analyzed. The analysis revealed that 32.1% of silages fell outside the reference pH range, indicating potential aerobic instability. While Salmonella and Campylobacter were not detected, Clostridium spp. were highly prevalent (81.0%), and C. perfringens was confirmed in 24.9% of samples. Listeria species occurred in 2.9% of silages, with L. innocua being the most frequent isolate. Statistical analysis showed that organic fertilization was significantly linked to specific C. perfringens toxotypes, though it did not increase the overall microbial burden. Conversely, microbial inoculation generally reduced the counts of several undesirable bacteria, although these differences were not statistically significant across all parameters. High pH values and significant contamination with Clostridium, B. cereus, and fungi remain critical challenges for silage safety. The results underscore the necessity for improved agricultural practices—specifically the minimization of soil and manure contamination during harvest—and the broader adoption of microbial inoculation to ensure the microbiological stability of fermented forage. Full article

Campylobacter is the primary bacterial cause of foodborne gastroenteritis globally. While international standards recommend a tiered approach for detection, emerging evidence suggests that selective protocols may introduce species-specific recovery biases. This systematic review and meta-analysis evaluated the diagnostic performance of established culture-based detection protocols across diverse food matrices. Following PRISMA 2020 guidelines, we searched multiple databases for studies reporting 2 × 2 diagnostic accuracy data through October 2024. Ten studies comprising 43 method comparisons and 4599 samples met the inclusion criteria. The overall pooled sensitivity was 95.8% (95% CI: 93.6–97.4%), and the specificity was 90.2% (95% CI: 86.8–92.9%). Even with a limited number of comparisons (n = 2), direct culture demonstrated high diagnostic sensitivity (99.1%) and significantly faster turnaround times. Crucially, selective enrichment exhibited a profound species-specific bias: C. jejuni showed 59.4 percent lower recovery than C. coli in Bolton broth, likely due to differential polymyxin B susceptibility. These findings highlight the importance of context-dependent method selection within the ISO 10272-1:2017 framework, suggesting that direct culture (Procedure C) should be prioritized for high-contamination matrices to ensure unbiased recovery of C. jejuni. Large-scale multicenter validation is warranted to confirm these exploratory findings for direct culture. Full article

Campylobacter (C.) jejuni and C. coli are common zoonotic bacteria in pigs, which typically act as asymptomatic carriers. However, the effects of Campylobacter colonisation on the porcine intestinal microbiota and metabolome remain poorly understood. This study investigated microbiome and metabolome alterations associated with co-colonisation by C. jejuni and C. coli in the different intestinal segments of pigs. Thirty-two weaned piglets were assigned to a control group and a group inoculated with C. coli ST5777/CT828 and C. jejuni ST122/CT206. Four weeks post inoculation, jejunal and caecal contents were analysed for Campylobacter counts, metabolite profiles and microbial composition. All animals remained clinically healthy. Both Campylobacter species colonised the jejunum and caecum, with higher C. coli counts in the caecum. Campylobacter-colonised pigs showed significantly altered metabolite profiles, including reduced cysteine and urea and increased glycine in the jejunum, as well as elevated 3-hydroxybutyrate levels in the caecum. In contrast, short-chain fatty acid concentrations in the caecum were unaffected by infection. Microbiota analysis revealed a significant reduction in caecal alpha diversity, whereas jejunal diversity remained unchanged. Infected pigs exhibited increased relative abundances of Lactobacillaceae and Bifidobacteriaceae and a decreased abundance of Pseudomonadota, including Enterobacteriaceae. In conclusion, Campylobacter co-colonisation induces distinct microbiome and metabolome alterations in pigs despite the absence of clinical disease. These findings highlight complex host-microbiota–pathogen interactions that may be relevant for future Campylobacter control strategies in pig production. Full article

Fluoroquinolone antibiotics, like danofloxacin, are considered as crucial veterinary drugs due to their high antibacterial potential, a broad spectrum of activity and good pharmacological properties. However, owing to the widespread use of this group of pharmaceuticals, microbial resistance to them is becoming a serious worldwide concern. In the present study, novel silver and copper complexes of danofloxacin were prepared and characterized using ¹H NMR, ¹⁹F NMR and IR spectroscopy, ESI-MS spectrometry, and elemental analysis. The antimicrobial properties of the obtained complexes were determined against selected bacterial and fungal strains, including yeast and conidia-forming fungi. Additionally, toxicities of danofloxacin metal-based complex solutions were assessed toward eukaryotic cells. The obtained results indicate that silver(I) and copper(II) complexes of danofloxacin exhibit good antimicrobial activity against bacteria that are important from the veterinary point of view, like Listeria monocytogenes or Campylobacter jejuni, in concentrations which are not cytotoxic. The MBC values of metal-based danofloxacin complexes for the mentioned strains were 1.5 times lower than those obtained for danofloxacin. Additionally, the solution of the novel silver–danofloxacin complex was found to have a fungicidal effect against the studied Candida and Aspergillus strains. Full article

To explore the appropriate supplementation rate of yeast culture (YC) in Kazakh sheep during fattening, the effects of different YC supplementation rates on rumen fermentation parameters and microbial community were studied through in vitro rumen fluid fermentation experiments. A 0.40 g high-concentrate diet was used as the fermentation substrate, and five groups were added with YC at 0% (CK), 1.25% (YC1), 2.5% (YC2), 3.75% (YC3) and 5% (YC4) of dietary dry matter, respectively. Anaerobic fermentation was carried out for 48 h in 60 mL fermentation broth. The results showed that the 48 h GP and microbial crude protein (MCP) concentration in all YC supplementation groups were significantly higher than those in the CK group (p < 0.05). The concentrations of total volatile fatty acids (TVFA) and propionate in the YC1 and YC2 groups were significantly increased and the A/P ratio in the two groups was significantly decreased (p < 0.05). The Multi-factor Comprehensive Evaluation Index (MFAEI) calculation indicated that 1.25% was appropriate. The YC1 and YC2 groups significantly increased the richness and diversity of rumen bacterial communities (Chao1 and Shannon indices, p < 0.05), and significantly increased the relative abundance of Bacteroidota and NK4A214_group (p < 0.05), while significantly decreasing the relative abundance of the potential pathogenic bacterium Campylobacter (p < 0.05). Ustilago abundance was significantly suppressed in all the YC-supplemented groups (p < 0.05). The most effective YC supplementation rate among the tested doses was 1.25% according to the MFAEI and key microbial indicators. The results suggest that dietary supplementation of 1.25% YC (dry matter basis) may beneficially modulate rumen fermentation parameters under in vitro conditions, providing a reference for further in vivo studies on its application in fattening Kazakh sheep. Full article