Search Results (339)

Background: Domestic refrigeration and egg handling are key factors in ensuring household food safety. Inadequate temperature control and poor hygiene in refrigerators can promote the survival and growth of foodborne pathogens. This study aimed to (i) characterize refrigerator temperature profiles and surface microbial contamination and (ii) screen eggs and egg-storage areas for the presence of Salmonella spp. and Campylobacter spp. Methods: Fifty domestic refrigerators were monitored twice in 2024 and 2025 in Porto, Portugal. The temperatures were continuously logged on the lowest shelf, which was swabbed for microbiological analysis. Surface hygiene was evaluated using total viable counts (TVC), Enterobacteriaceae, and Escherichia coli enumerated following ISO methods. Detection of pathogens Listeria monocytogenes, Salmonella spp., and Campylobacter spp. was performed using real-time PCR. Eggs (n = 92 in 2024; n = 88 in 2025), and domestic egg storage areas (total n = 76) were screened for Salmonella and Campylobacter. Results: The mean refrigerator temperatures were 6.0 ± 0.5 °C in 2024 and 6.1 ± 0.5 °C in 2025; 44% and 50% of the units, respectively, exceeded the recommended 6 °C threshold. In 2025, 31 (62%) and 33 (66%) refrigerators showed higher TVC and Enterobacteriaceae counts compared to 2024, whereas E. coli was only detected sporadically. L. monocytogenes, Salmonella spp., or Campylobacter spp. were not recovered from the refrigerator surfaces. Likewise, Salmonella and Campylobacter were not detected in any of the eggs or egg-storage sites. Indicator microorganism’s counts were not associated with the mean temperature. Conclusions: The absence of correlation between ΔT and Δ microbial counts suggests that behaviour-driven hygiene factors, rather than the relatively small year-to-year temperature differences observed, are more influential in determining household bioburden. Maintaining refrigerator temperatures ≤ 6 °C together with simple hygiene practices remains essential for reducing household food safety risks. Full article

Blood culture remains the gold standard for identifying bloodstream infections caused by bacteria and fungi. Isolation of the culprit microorganism onto agar plates also facilitates antimicrobial susceptibility testing. The purpose of this study was to determine the contamination rates, pathogen profile, and antimicrobial resistance in a secondary healthcare setting in a two-year timeframe. In this study, data regarding blood cultures of the years 2023 and 2024 were retrospectively analyzed to address the above questions. Blood cultures were incubated for seven days before being discarded as negative. The percentage of positive blood cultures for both years was 14.3%. Most positive cultures contained Gram-positive cocci, with a prevalence of coagulase-negative Staphylococci. In descending order, 72.72% were coagulase-negative Staphylococci, 15.15% were Staphylococcus aureus, and 12.12% were Streptococci. One strain of S. aureus was methicillin-resistant (MRSA), and one strain of Enterococcus faecium was vancomycin-resistant (VRE). Of the Gram-negative rods, 78.3% were Enterobacterales. Of these, Escherichia coli, Klebsiella pneumoniae, and Proteus mirabilis were the top pathogens. The remainder comprised eight strains of Pseudomonas aeruginosa, four strains of Acinetobacter baumannii (one pandrug-resistant), three strains of Stenotrophomonas maltophilia, one strain of Sphingomonas paucimobilis, and one strain of Campylobacter jejuni. The isolated fungi comprised Candida parapsilosis, Candida glabrata, and Candida tropicalis. Of the isolated Escherichia coli strains, 39.5% were resistant to ciprofloxacin regardless of origin (outpatient or hospitalized patients). Outpatient samples were taken in a Hemodialysis Unit that collaborates with our laboratory, obtained from patients with fever or other signs of infection. Distinguishing true bacteremia from contamination remains challenging. The contamination rate in our study was quite high at 5.3%. Since there is no dedicated phlebotomy team in our healthcare setting, in light of our results, educational courses have been conducted to demonstrate the best practices for sample collection. Full article

Antimicrobial resistance (AMR) in foodborne pathogens poses a major threat to global public health and food safety. Using 9393 U.S. isolates of Salmonella enterica, Campylobacter jejuni, and Escherichia coli/Shigella collected from poultry, cattle, and swine between 2015 and 2025 and archived in the NCBI Pathogen Isolates Browser, we applied multivariate statistical analysis to characterize antimicrobial resistance patterns in isolates showing resistance to one to six antimicrobials (AMR-1 to AMR-6). Six antimicrobials—tetracycline, streptomycin, sulfisoxazole, ampicillin, nalidixic acid, and ciprofloxacin—were identified through PCA-guided clustering and frequency profiling as the principal axes of co-resistance across pathogens. Tetracycline emerged as a foundational driver of multidrug resistance, while C. jejuni contributed almost exclusively to single-drug resistance and Salmonella enterica dominated higher-order AMR categories, reflecting species-specific ecological and genomic constraints. Gene analyses revealed a progressive, modular accumulation of resistance determinants, led by efflux pumps (mdsA, mdsB), tetracycline genes (tetA/B/O), aminoglycoside-modifying enzymes, sulfonamide genes (sul1/sul2), quinolone resistance determinants (gyrA, acrF, mdtM), and β-lactamases (bla_EC, bla_OXA, bla_CTX). Together, these results demonstrate that multidrug resistance in U.S. foodborne pathogens evolves through coordinated gene–drug–pathogen interactions rather than isolated events, underscoring the need for integrated surveillance and targeted stewardship strategies focused on the dominant antimicrobials and high-risk foodborne pathogens. Full article

Ensuring optimal food hygiene is essential for food safety and preventing foodborne illness, although the importance of food hygiene is often overlooked in the household kitchen setting. Adequate, good hygiene practices in the domestic environment are equally important as their implementation in any other food preparation environment, like in the food industry. The current review encompasses research data on the prevalence and isolation of major foodborne pathogenic bacteria (Campylobacter, Salmonella, Listeria monocytogenes, Staphylococcus aureus, Escherichia coli pathotypes, and Clostridium perfringens) from household kitchen equipment, as well as food cleaning utensils used in the kitchen, such as sponges, brushes, dishcloths, and hand towels. The most common bacterial pathogen present in the domestic environment is S. aureus. The latter can be transmitted orally, either via direct hand contact with contaminated kitchen surfaces and/or cleaning utensils, or indirectly through the consumption of contaminated food due to cross-contamination during food preparation (e.g., portioning prepared meat on the same cutting board surface and with the same knife previously used to cut fresh leafy vegetables). Moreover, research findings on the hygiene of food cleaning utensils demonstrate that (i) sponges have the highest microbial load compared to all other cleaning utensils, (ii) brushes are less contaminated and more hygienic than sponges, thus safer for cleaning cutlery and kitchen utensils, and (iii) kitchen dishcloths and hand towels positively contribute to cross-contamination since they are frequently used for multiple purposes at the same time (e.g., drying hands and wiping/removing excess moisture from dishes). Finally, the present review clearly addresses the emerging issue of antimicrobial resistance (AMR) in bacterial pathogens and the role of the domestic kitchen environment in AMR dissemination. These issues add complexity to foodborne risk management, linking household practices to broader AMR stewardship initiatives. Full article

The Indo-Pacific region hosts several annual military exercises that involve the deployment of thousands of U.S. and partner-nation military personnel. Respiratory and diarrheal diseases pose a significant health risk to exercise participants and represent a substantial portion of medical encounters and lost duty days. We conducted surveillance for respiratory and diarrheal illness at the Cobra Gold and Balikatan military exercises in Thailand and the Philippines from 2023–2025. Through coordination with health providers in the field, military personnel that reported acute symptoms were asked to provide a nasopharyngeal swab or stool sample. These samples were transported to a field lab and tested by PCR for common respiratory and diarrheal pathogens. Follow-up analyses included bacterial culture, antimicrobial susceptibility testing, and viral whole-genome sequencing. From 84 respiratory and 61 diarrheal samples analyzed, we found that respiratory illness was primarily attributed to rhinoviruses/enteroviruses (23%), common coronaviruses (21%), and SARS-CoV-2 (11%) while diarrheal disease was attributed to a high rate of diarrheagenic E. coli (73%) and norovirus (20%), followed by Salmonella spp. (18%) and Campylobacter spp. (13%). Our findings highlight the distinct etiologies of respiratory and diarrheal disease in military field settings and demonstrate the feasibility of conducting real-time infectious disease surveillance in operational environments. Full article

Postbiotics, which are preparations of inanimate microorganisms and their components, have emerged as a promising functional ingredient in animal health and nutrition. Postbiotics are primarily composed of microbial cell fractions, metabolites, enzymes, vitamins, polysaccharides, and short-chain fatty acids. Unlike probiotics, postbiotics do not contain live microorganisms, which strengthens their greater stability and safety in feed/food formulations. Postbiotics offer several beneficial effects, including antioxidant, anti-inflammatory, immune-modulatory, and antimicrobial actions. They enhance antioxidant enzymes, neutralize reactive oxygen species, and inhibit lipid peroxidation, thereby protecting tissues from oxidative damage. Postbiotics also inhibit pro-inflammatory molecules like TNF-α and IL-6, while enhancing the anti-inflammatory cytokine IL-10, promoting the maturation and function of immune cells, and increasing secretory IgA production. They suppress a variety of pathogenic bacteria, including Escherichia coli, Salmonella, Staphylococcus, Campylobacter, etc., both in vitro and in vivo. Moreover, they increase beneficial gut bacteria and improve the digestion and integrity of the intestine. This article outlines the beneficial effects of postbiotics in animals including poultry, swine, canine, feline, horses, and ruminant animals, either as feed/food or as a supplement. The integration of postbiotics into animal feed improves growth performance, feed conversion ratios, and disease resistance in animals. Thus, the multifunctional benefits of postbiotics make them a valuable tool for healthy companion animals and sustainable livestock production, supporting both animal welfare and productivity without the drawbacks associated with antibiotic growth promoters. Full article

Objectives: This observational and cross-sectional study investigated differential associations between entero-invasive and non-entero-invasive enteric pathogens and HIV infection, considering socioeconomic, clinical and immunological aspects. In a Ghanaian population with a high prevalence of enteric pathogens, stool samples from people living with HIV (PLWH) were screened for Salmonella spp., Shigella spp./EIEC (enteroinvasive Escherichia coli), and Campylobacter jejuni as entero-invasive bacteria, for enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), and enteroaggregative E. coli (EAEC) as non-entero-invasive bacteria. Arcobacter butzleri, with uncertain enteropathogenicity, was also included. Methods: Stool samples from PLWH (with and without antiretroviral therapy) and HIV-negative controls were analyzed by real-time PCR for the presence and quantity of the selected enteropathogens. Results were correlated with socioeconomic, clinical, and immunological parameters. Results: The presence of Shigella spp. /EIEC in stool was both qualitatively and quantitatively associated with reduced CD4+ T lymphocyte counts and was qualitatively associated with clinically apparent diarrhea. EAEC showed a weak positive association with HIV infection, supported by a negative correlation between EAEC DNA quantity and CD4+ T lymphocyte counts. EPEC colonization was associated with HIV negativity, higher CD4+ T lymphocyte counts, and lower socioeconomic status. Abundance of Salmonella enterica was associated with clinically apparent diarrhea. Conclusions: This explorative, hypothesis-forming study suggests species- or pathovar-specific associations between enteric bacterial pathogens and HIV-related immunosuppression. Observed relationships with clinically apparent diarrhea largely align with findings from sub-Saharan African children, except for a more pronounced association between diarrhea and Salmonella in this cohort. Full article

Wild boars, widely distributed across natural, agricultural, and urban landscapes, represent an ideal sentinel species for monitoring the emergence and spread of antimicrobial resistance (AMR) at the human–wildlife–livestock interface within the One Health framework. This review summarizes current knowledge on the prevalence, diversity, AMR, and epidemiological significance of major enteric pathogens isolated from wild boars in the European Union, with particular attention to their potential role in AMR dissemination. Numerous studies have reported variable prevalence rates for Salmonella spp., Yersinia enterocolitica, Shiga toxin-producing Escherichia coli (STEC), and Campylobacter spp. High prevalence rates has been observed in fecal samples—35% for Salmonella, 27% for Y. enterocolitica and STEC, and 66% for Campylobacter—highlighting the role of wild boars as carriers and the associated risk of carcass contamination during slaughter. Tonsils represent a key niche for Y. enterocolitica, with prevalence reaching 35%. Several studies have identified resistance to antimicrobials classified by the World Health Organization as critically important or high priority for human medicine, including fluoroquinolone-resistant non-typhoidal Salmonella spp. and third-generation cephalosporin-resistant Y. enterocolitica, raising notable public health concerns. Despite increasing interest, most available studies remain descriptive and geographically limited, providing limited insight into AMR acquisition and transmission pathways in wild boars. New approaches—such as resistome analyses and epidemiological cut-off values—offer added value to distinguish wild-type from acquired-resistant strains and to better understand AMR dissemination dynamics. Integrating wildlife into One Health surveillance systems is essential to capture the full complexity of AMR spread. Full article

Background/Objectives: The global emergence of carbapenem resistance is a major public health concern. Campylobacter jejuni and Campylobacter coli, key zoonotic agents causing human campylobacteriosis, are mainly isolated from poultry, their primary host. Their increasing resistance in animals and humans highlights the risk of gene transfer. This study investigates the molecular mechanisms of carbapenem resistance in 287 avian Campylobacter spp. isolates from Tunisia within a One Health approach. Methods: Antibiotic susceptibility of 287 carbapenem-resistant isolates, including 147 C. jejuni and 140 C. coli, was determined according to CLSI. All isolates were screened by PCR for genes encoding the most reported carbapenemases, including VIM, IMP, NDM and OXA-48. Eleven multidrug-resistant (MDR)/carbapenem-resistant C. coli isolates were selected to determine their clonal lineage by Multilocus sequence typing (MLST). Results: All isolates were susceptible to imipenem, but resistance to meropenem and ertapenem were observed in 60.71% and 35.71% of C. coli isolates, respectively, versus 13.6% in C. jejuni for each antibiotic. The bla_VIM, bla_NDM and bla_OXA-48 genes were detected in 15, 8, and 19 of the 20 C. jejuni isolates, respectively. However, for C. coli, 53, 12, and 15 isolates harbored bla_VIM, bla_NDM and bla_OXA-48 genes, respectively. The eleven (MDR)/carbapenem-resistant C. coli isolates belonged to a unique ST sequence type ST13450. Conclusions: We report for the first time the emergence of bla_VIM, bla_NDM, and bla_OXA-48 genes in Campylobacter spp. isolates of poultry origin highlighting possible horizontal transfer of these genes to pathogenic Gram-negative bacteria of the poultry’s microbiota. Full article

The pond slider (Trachemys scripta) is native to the southeastern United States but has been introduced all around the world, including to the Canary Islands (Spain), along with other less-common exotic freshwater turtles such as Graptemys pseudogeographica, Mauremys spp., and Pseudemys peninsularis. The aim of this study was to determine the presence of pathogenic bacteria in these animals and to evaluate the associated health risks for humans and local fauna. For this purpose, cloacal samples from 42 specimens collected on the islands of Tenerife and Gran Canaria (Canary Islands) were analyzed for potentially zoonotic bacteria using selective culture media and PCR. Non-tuberculous mycobacteria were the most isolated pathogen (57.9%), followed by Yersinia enterocolitica (42.1%) and Escherichia coli carrying stx and/or eae genes (33.3%). Salmonella spp. was detected in 31.0% of the chelonians, identifying Salmonella Typhi and Salmonella Typhimurium serotypes. Staphylococcus spp. showed a prevalence of 21%, mainly Staphylococcus aureus along with one antibiotic-resistant Staphylococcus hominis isolate. Pseudomonas spp. were found in 10.1% of samples, although only one isolate corresponded to Pseudomonas aeruginosa. Campylobacter spp. and Vibrio spp. were detected at low frequencies (<10%), and Listeria monocytogenes was not identified. Overall, the results indicate that aquatic turtle populations in the Canary Islands pose notable health risks, especially for animal handlers and people with compromised immune systems. Full article

Background/Objectives: Goat and sheep farming is an important agro-economic resource in Benin. However, their milk is both underutilized and insufficiently characterized, which limits the development of innovative dairy products and raises concerns about its safety. Against this backdrop, our pioneering study set out to investigate, for the first time in Benin and using an advanced metagenomic approach, the microbial diversity present in goat and sheep raw milk. The aim was to lay the groundwork for safer and more efficient dairy valorization. Methods: To achieve this, metagenomic DNA was extracted from 20 pooled milk samples representing both animal species, followed by shotgun sequencing. Results: Analyses revealed seven dominant phyla: Bacillota (17.44–27.23%), Pseudomonadota (12.39–15.55%), Campylobacterota (3.65–5.29%), Actinomycetota (1.47–6.03%), Spirochaetota (1.14–2.02%), Apicomplexa (0.28–0.50%), and Bacteroidota (0.17–0.22%) in the raw milk of both species. However, their proportions differ. Bacillota, which was the most dominant in both types of milk, was significantly more abundant in goat (27.23 ± 5.33) than in sheep milk (17.44 ± 8.44). In sheep milk, Enterobacteriaceae (11.36 ± 5.79) were the most predominant family, followed by Streptococcaceae (5.57 ± 2.29) and Staphylococcaceae (4.51 ± 3.63). Goat milk, on the other hand, presents a different hierarchy. Streptococcaceae (6.65 ± 2.19) and Staphylococcaceae (6.43 ± 2.33) were the most abundant families, surpassing Enterobacteriaceae (5.33 ± 1.66). The genus Escherichia was the most abundant in sheep milk (6.18 ± 5.33). The genera Staphylococcus (4.50 ± 3.63) and Streptococcus (5.05 ± 1.98) were also present. In contrast, in goat milk, the genera Streptococcus (6.54 ± 2.35) and Staphylococcus (6.42 ± 2.32) were the most dominant, while the average abundance of Escherichia was much lower (1.98 ± 1.28). In terms of species, Sheep milk was dominated by Escherichia coli (6.14 ± 5.28) and Staphylococcus aureus (5.17 ± 2.28) while Klebsiella pneumoniae (2.82 ± 1.72), Streptococcus pneumoniae (1.92 ± 1.36), and Campylobacter coli (1.52 ± 1.27) were also found. In addition to a relatively high abundance of Staphylococcus aureus (6.40 ± 2.45), goat milk was characterized by the presence of Corynebacterium praerotentium (5.32 ± 2.28) and Clostridium perfringens (3.39 ± 2.09). Additional pathogens identified included Clostridioides difficile (1.17–2.00%), Clostridium botulinum (0.27–0.43%), Listeria monocytogenes, Mycobacterium tuberculosis, Helicobacter pylori (0.36–0.62%), Salmonella enterica (0.22–0.26%). As for fungi, Ascomycota were predominant, with the presence of Aspergillus fumigatus, Saccharomyces cerevisiae, Trichophyton mentagrophytes, and Candida auris. Moreover, lactic acid bacteria with technological interest such as Oenococcus oeni (0.60–0.97%), Levilactobacillus namurensis (0.25–0.44%), Lactobacillus agrestimuris, and Lacticaseibacillus rhamnosus were also detected. Conclusions: These findings provide essential insights into the technological potential and health risks associated with these milks, which are key to developing safer and more efficient local dairy value chains. Full article

Bacteriophages, or phages, are viruses that infect and kill specific bacteria, offering a promising alternative to antibiotics in livestock production. With growing concerns over antibiotic resistance, phages have gained renewed interest due to their ability to target harmful pathogens without disturbing beneficial gut microbiota. This review explores the application of dietary phage supplementation in monogastric animals, particularly pigs and poultry. In pigs, phage use has demonstrated beneficial effects such as improved growth performance, enhanced gut health, and reduced infections from Salmonella and E. coli. Various delivery methods, including feed and water supplementation, have been studied, with microencapsulation showing promising results for stability and effectiveness. Similarly, in poultry, phages have been successfully used to control pathogens like Salmonella, Campylobacter, and avian pathogenic E. coli, improving gut health, immunity, and overall performance. Several commercial phage products are already in use, demonstrating both safety and efficacy. Despite these advantages, challenges such as a narrow host range, bacterial resistance, and regulatory limitations remain. Therefore, further research is necessary to understand phage–host interactions, optimize delivery strategies, and evaluate long-term effects under normal and disease-free conditions. This review highlights the potential of bacteriophages as safe, targeted, and sustainable alternatives to antibiotics in monogastric animal production, contributing to improved animal health and reduced antibiotic use. Full article

Background/Objectives: In this work, a series of six new indazole-benzimidazole hybrids (M1–M6) were designed, synthesized, and fully characterized. The design of these compounds was based on the combination of two pharmacophoric units, indazole and benzimidazole, both known for their broad spectrum of biological activities. Methods: The molecular hybridization strategy was planned to combine these scaffolds through an effective synthetic pathway, using 6-nitroindazole, two 2-mercaptobenzimidazoles, and 1,3- or 1,5-dihaloalkanes as key precursors, affording the desired hybrids in good yields and with enhanced biological activity. Quantum chemical calculations were performed to investigate the structural, electronic, and electrostatic properties of M1–M6 molecules using Density Functional Theory (DFT) at the B3LYP/6-311++G(d,p) level. The antimicrobial activity efficacy of these compounds was assessed in vitro against four Gram-positive bacteria (Staphylococcus aureus, Enterococcus faecalis, Bacillus cereus, and Lactobacillus plantarum), four Gram-negative bacteria (Salmonella enteritidis, Escherichia coli, Campylobacter coli, Campylobacter jejuni), and four fungal strains (Saccharomyces cerevisiae, Candida albicans, Candida tropicalis, and Candida glabrata) using ampicillin and tetracycline as reference standard drugs. Results: Among the series, compound M6 exhibited remarkable antimicrobial activity, with minimum inhibitory concentrations (MIC) of 1.95 µg/mL against S. cerevisiae and C. tropicalis, and 3.90 µg/mL against S. aureus, B. cereus, and S. enteritidis, while the standards Ampicillin (AmB) (MIC ≥ 15.62 µg/mL) and Tetracycline (TET) (MIC ≥ 7.81 µg/mL) exhibited higher MIC values. To gain molecular insights into the compounds, an in silico docking study was performed to determine the interactions of M1–M6 ligands against the antimicrobial target beta-ketoacyl-acyl carrier protein (ACP) synthase III complexed with malonyl-COA (PDB ID: 1HNJ). Molecular modeling data provided valuable information on the structure-activity relationship (SAR) and the binding modes influencing the candidate ligand-protein recognition. Amino acid residues, such as Arg249, located in the solvent-exposed region, were essential for hydrogen bonding with the nitro group of the 6-nitroindazole moiety. Furthermore, polar side chains such as Asn274, Asn247, and His244 participated in interactions mediated by hydrogen bonding with the 5-nitrobenzimidazole moiety of these compound series. Conclusions: The hybridization of indazole and benzimidazole scaffolds produced compounds with promising antimicrobial activity, particularly M6, which demonstrated superior potency compared to standard antibiotics. Computational and docking analyses provided insights into the structure–activity relationships, highlighting these hybrids as potential candidates for antimicrobial drug development. Full article

The consumption of raw milk has been demonstrated to carry a potential risk of transmission of Campylobacter spp., with Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) being the major causes for foodborne gastroenteritis cases. The present study assessed the prevalence and species distribution of Campylobacter spp. in 633 raw milk samples collected over a one-year climatic cycle from small, medium, and large producers in Pichincha and Manabí, Ecuador. Samples were augmented and analyzed by qPCR for Campylobacter spp., while species identification was performed by duplex PCR and confirmed by 16S rDNA sequencing. The average prevalence of Campylobacter spp. was 49.9% (316/633), with a higher detection rate in Manabí (57.6%, 182/316) compared to Pichincha (42.4%, 134/316). C. coli was the most prevalent species, accounting for 46.2% (146/316) of the cases, followed by C. jejuni at 23.1% (73/316), co-contaminations at 13.3% (42/316), and non-identified Campylobacter at 44.0% (139/316). Phylogenetic analysis was employed to confirm species identity, thereby confirming the presence of Campylobacter fetus and Campylobacter lari. The increased diversity and frequency of isolates in Manabí, particularly during periods of elevated temperature, imply that coastal environmental conditions and production practices promote the persistence of bacteria. The findings of this study indicate a high prevalence of Campylobacter in Ecuadorian raw milk, posing a significant health risk to the population and underscoring the need for enhanced hygiene practices and continuous monitoring to mitigate public health risks. Full article

Campylobacter spp. are significant zoonotic pathogens, increasingly recognized for their role in the transmission of antimicrobial resistance (AMR) between animals and humans. This study aims to determine the occurrence, antimicrobial resistance profiles, and characterization of multidrug resistance indices of Campylobacter spp. isolated from domestic pets in Kelantan, Malaysia. Methods: Rectal swabs (n = 150) were collected from both healthy and diarrheic cats. Campylobacter spp. were isolated and confirmed by PCR, and antimicrobial susceptibility was assessed using the disk diffusion method. Results: Campylobacter spp. were detected in 35.3% of cats (53/150; SE = 0.04; 95% CI: 28.1–43.3%), with C. upsaliensis identified as the predominant species (33.3%; SE = 0.05; 95% CI: 24.5–43.6%), followed by C. jejuni (17.8%; SE = 0.04; 95% CI: 11.3–26.9%) and C. coli (7.8%; SE = 0.03; 95% CI: 3.8–15.2%). Isolates exhibited high resistance rates to amoxicillin (90.6%), ampicillin (81.1%), tetracycline (67.9%), erythromycin (62.3%), and sulphonamides (54.7%). Conclusion: The study reveals a substantial prevalence of Campylobacter spp. and notable levels of antimicrobial resistance among feline isolates, highlighting the zoonotic threat in Malaysia. These findings emphasize the importance of integrated surveillance and prudent antimicrobial stewardship under a One Health framework. Full article