Foodomics Approaches to Facilitate the Verification of the Authenticity of Foods: A Possible Strategy to Screen,Validate, and Standardize Food Matrices / Enfoques de Foodomics para facilitar la verificación de la autenticidad de los alimentos: una posible estrategia para evaluar, validar y estandarizar matrices alimentarias

The complexity of globalization, including the global food trade market, has the side effect that various raw foodstuffs are vulnerable to intentional and unintentional adulteration. However, food validation and standardization approaches are still unclear and challenging and need to be explored. Through this opinion article, the author would like to introduce a foodomics approach (Food, -Omics) to facilitate integrated food authenticity verification through biosensors. This approach is potentially suitable and offers more valuable accuracy as it combines biological analysis methods spanning genomics, transcriptomics, proteomics, and metabolomics. Meanwhile, several subdisciplines of Foodomics, such as metallomics, volatomics, and lipidomics, which are considered feasible to facilitate the verification of food authenticity, are also explored in this critical opinion. Foodomics consists of four main omics technologies, namely genomics, transcriptomics, proteomics, and metabolomics. This is an integration of promising approaches to provide standardized food matrices, thus becoming the most likely strategy to verify the authenticity of food. However, after trying to uncover this food authentication problem and provide a Foodomics approach, we felt the need for synergies in building a database capable of storing food matrices in the form of unique genes, bioactive peptides, and secondary metabolites. We hope that through this opinion article, the target database can be formed, although databases such as MEDLINE and PubChem have provided this data facility. In particular, we suggest the development of nanobiosensors that should undoubtedly be environmentally friendly and portable (making use of smartphones) and creating a cloud database capable of storing food matrices in the form of unique genes, bioactive peptides, and secondary metabolites, integrated with smartphone biosensors.(AU)

Biliary protozoa in a dog with acute cholangiohepatitis fed a raw food diet.

A 1-year 11-month intact female Alaskan Malamute fed a raw food diet was referred to the Queen Mother Hospital for Animals for further investigation of hyporexia and increased hepatobiliary enzyme activities. Clinicopathological and imaging findings were consistent with cholangiohepatitis, with coccidial zoites identified on bile cytology. Polymerase chain reaction and amplicon sequencing from the bile identified Hammondia heydorni, a Sarcocytid coccidial protozoa with an obligate 2-host life cycle. The dog was treated with clindamycin, marbofloxacin, ursodeoxycholic acid (UDCA) and S-adenosylmethionine/silybin with complete clinical and biochemical resolution documented after 6 weeks. Infection with Hammondia spp. should be considered in patients receiving raw food diets in which coccidial zoites are identified in the bile, but the pathogenic potential of this organism is unknown and the possibility of its presence as a commensal cannot be discounted.

Determination of water in organic solvents and raw food products by fluorescence quenching of a crystalline vinyl-functionalized COF.

Covalent organic frameworks (COFs) with good chemical stability, flexible chemical functionalization, tunable pore sizes, and high specific surface areas have been increasingly employed in the field of fluorescence sensing. In this work, a crystalline vinyl-functionalized COF TzDa-V was facilely prepared through a room-temperature synthetic method via condensation reaction between 4,4',4″-(1,3,5-triazine-2,4,6-triyl)trianiline (Tz) and 2,5-diallyloxyterephthalaldehyde (Da-V). The intermolecular charge transfer (ICT) effect endowed the TzDa-V with fluorescence characteristic, and it was sensitive to trace water and can be quenched due to the disruption of ICT process by water. On this base, the prepared COF TzDa-V with excellent chemical/thermal stability was applied to sensing of trace water in common organic solvents such as DMF, acetone, THF, and ethyl acetate with rapid response (less than 10 s), satisfactory sensing range (0.5-18% water in DMF, 0.5-15% water in acetone, 0.5-16% water in THF, 0.5-5% in ethyl acetate, v/v), and high sensitivity. The limits of detection for water in DMF, acetone, THF, and ethyl acetate were 0.0497%, 0.0590%, 0.0502%, and 0.0766% (v/v), respectively. The proposed probe was successfully used for the detection of trace water in food products such as salt and sugar. The COF TzDa-V would be a good candidate for application in water sensing.

Evaluation of canine raw food products for the presence of extended-spectrum beta-lactamase- and carbapenemase-producing bacteria of the order Enterobacterales.

OBJECTIVE: To assess the potential contamination of commercial raw dog food products with bacteria of the Enterobacterales order that produce extended spectrum beta-lactamase (ESBL) and carbapenemase enzymes, determine risk factors for contamination, and understand isolate genetic diversity. SAMPLES: A total of 200 canine raw food products. METHODS: Products were cultured on selective chromogenic agar following enrichment steps. Whole-genome sequencing was performed for isolates that were confirmed to produce an ESBL. Isolates were characterized by antimicrobial resistance genes, and multilocus sequences typing, and compared to other isolates in the NCBI database for clonality. Preservation method and protein sources were assessed as potential risk factors for contamination with ESBL and carbapenemase-producing bacteria of the Enterobacterales order. RESULTS: No carbapenemase-producing Enterobacterales (CPE) were identified, but ESBL-producing Enterobacterales bacteria were isolated from 20/200 products (10.0%; 95% CI, 7.3 to 16.5%), all of which were frozen. Pork-derived protein source products were 8.1 times (P = .001; 95% CI, 2.53 to 26.2) more likely to carry ESBL-producing Enterobacterales bacteria than other protein sources. WGS analysis confirmed the presence of ESBL genes in a total of 25 distinct isolates (19 Escherichia coli, 5 Klebsiella pneumoniae, and 1 Citrobacter braakii). Genes encoding CTX-M type ESBL enzymes were the most common (24/25 isolates, 96.0%) with blaCTX-M-27 being the most common allele (8/25, 32.0%). CLINICAL RELEVANCE: Frozen, raw food products may serve as a route of transmission of ESBL-producing Enterobacterales bacteria to companion animals. Veterinarians should advise owners about the risks of raw food diets, including potential exposure to antimicrobial-resistant bacteria.

Nutritional Intake and Biomarker Status in Strict Raw Food Eaters.

Following a strict raw food diet (primarily based on fresh fruit and raw vegetables, waiving any consumption of heated or processed food) has the risk of undersupply of energy and certain macro- and micronutrients. In this cross-sectional study, we compared 16 non-smoking strict raw food eaters (5 women and 11 men, age 44.6 ± 12.3 years, duration of following the diet 11.6 ± 10.8 years) with the non-smoking participants (32 vegans, 27 omnivores) of the "Risk and Benefits of a Vegan Diet" (RBVD) study. We investigated body composition, dietary intake from 3-day weighed food records, and relevant fasting blood and serum parameters. Food choice and dietary behavior were very heterogenic in raw food eaters. They had lower mean values of BMI and percentage of body fat than the respective RBVD participants. The same holds true for energy supply and intakes of protein, carbohydrate, calcium and iodine. Serum levels revealed lower levels of HDL cholesterol, triglycerides, zinc, and vitamin D3. The raw food eaters with (n = 9) and without (n = 7) supplementation of vitamin B12 had median vitamin B12 levels of 399 and 152 ng/L, respectively. Accordingly, eight raw food eaters (50%) had homocysteine levels above 12 µmol/L. The study allows a close look at strict raw food eaters with respect to possible dietary deficiencies, but also provides insights into motivations and daily life.

Qualidade do leite cru refrigerado avaliado por diferentes laboratórios credenciados pelo MAPA / Quality of refrigerated raw milk evaluated by different laboratories accredited by MAPA

O objetivo deste trabalho foi analisar e correlacionar os resultados de qualidade do leite cru refrigerado de dez propriedades rurais de Viçosa (MG) obtidos por métodos de referências do Ministério da Agricultura, Pecuária e Abastecimento e por espectrofotômetros de luz infravermelha em três laboratórios (A, B e C) credenciados pelo mesmo órgão de fiscalização. As amostras de leite foram analisadas quanto à contagem bacteriana, contagem de células somáticas (CCS) e teores de gordura, proteína e extrato seco desengordurado (ESD). Não houve discordância de amostras de leite quanto aos teores de proteína e gordura. Porcentagens diferentes de discordância foram observadas entre os resultados dos laboratórios e método de referência quanto à CCS, contagem bacteriana e teores de ESD. Os teores médios de proteína dos laboratórios A e B e os teores médios de ESD de todos laboratórios não foram estatisticamente iguais (p<0,05) aos teores obtidos pelo método de referência. Os teores de gordura obtidos no laboratório B não tiveram correlação estatística (p>0,05) com os teores obtidos pelo método de referência. Observou-se que as CCS e contagens bacterianas médias de todos os laboratórios foram estatisticamente iguais (p>0,05) e correlacionadas (p<0,05) às contagens médias obtidas pelos métodos de referência. Conclui-se que os teores de proteína em dois laboratórios e ESD em todos os laboratórios não são equivalentes com os métodos de referência do MAPA. É importante que os laboratórios revejam a calibração dos seus equipamentos quanto à quantificação dos teores de sólidos do leite.

The objective of this study was to analyze and to correlate the results of refrigerated raw milk quality in ten farms of Viçosa (MG). The milk was analyzed by the Ministério da Agricultura, Pecuária e Abastecimento standard methods and by infrared spectrophotometers in three laboratories (A, B and C) accredited by the same institution. The milk samples were analyzed for bacterial counts, somatic cell count (SCC) and fat, protein and solids not fat (SNF) tenors. There were no disagreement milk samples as the protein and fat tenors. Different percentages of disagreement were observed between the results of laboratory and standard method as the SCC, bacterial count and SNF tenor. The protein mean tenor of A and B laboratories and the SNF mean tenor of all laboratories were not statistically equal (p<0.05) the tenors obtained by standard method. The fat tenors obtained in B laboratory were not statistically correlated (p>0.05) with tenors obtained by the standard method. It was observed that the means of SCC and bacterial counts of all laboratories were statistically the same (p>0.05) and correlated (p<0.05) to the means counts obtained by standard methods. It is concluded that the protein tenors in two laboratories and SNF in all laboratories are not equivalent with the reference methods of MAPA. It is important that laboratories review the calibration of the equipment on the quantification of milk solids tenors.

The influence of food processing methods on serum parameters, apparent total-tract macronutrient digestibility, fecal microbiota and SCFA content in adult beagles.

Food processing methods may influence the health of dogs. However, previous studies have mostly been based on a comparison of several commercial dog foods with different ingredients. In this study, eighteen adult beagles of the same age and health status (assessed by routine blood tests) were used in the experiments. This study analyzed the effects of the following different processing methods: raw, pasteurized, and high temperature sterilization (HTS) made with the same ingredients and nutrients (based on dry matter) on serum parameters, apparent total-tract macronutrient digestibility, fecal microbiota and short-chain fatty acid (SCFA) content in beagle dogs. The data showed, after a test lasting 56-days, the apparent digestibility (ATTD) of protein and fat in HTS food was 91.9%, which was significantly higher (P< 0.05) than that in dry food (89.2%, P < 0.05). The serum content of triglyceride increased in beagles fed HTS food (P < 0.05), and the number of neutrophils in beagles fed raw food and pasteurized food increased significantly (P < 0.05), and the platelet count in beagles fed raw food showed an increasing trend compared with the beagles fed HTS food. Different processing methods had an impact on the intestinal microbiota and SCFA of beagles; at least 14 genera were significantly affected by the food produced using different processing methods. In particular, the abundance of Allprevotella, Escherichia-Shigella and Turicibacter, and the total acid content were lower in beagles fed the raw diet, whereas Streptococcus, Collinsella, Bacteroides and Ruminococcus gnavus were more abundant following the HTS diet, and Lactococcus showed the highest abundance in beagles fed the pasteurized diet. This study showed that dog food produced by different processing methods affected the health of adult beagles.

Microbiological profile of raw refrigerated and processed bovine milk at dairy industries from Vale do Taquari, Rio Grande do Sul, Brazil

Milk is an essential food, widely consumed by the population. Brazil is one of the world's largest producers of milk. Milk quality is influenced by several factors in all its stages of production. The aim of this study was to determine the microbiological profile of refrigerated and processed raw bovine milk from industries in Vale do Taquari, state of Rio Grande do Sul, Brazil, using metagenomic analysis. A total of six samples were collected, one of refrigerated raw milk from the tanker truck, one of pasteurized milk and one of milk sterilized by the ultra-high temperature (UHT) process, in each of the industries. The identification of the milk microbiota was performed by sequencing the 16S rRNA gene. The results show that refrigerated raw milk has a greater number of microorganisms, followed by pasteurized milk and sterilized milk, successively. Processed milk showed the presence of beneficial microorganisms such as Streptococcus thermophilus and Streptococcus macedonicus. Nevertheless, even UHT milk showed the presence of microorganisms considered harmful, such as the Bacillus cereus group, Aeromonas dhakensis, Enterobacter bacterium and Acinetobacter haemolyticus. Metagenomics is a valuable tool for the thorough evaluation of the milk microbiota in order to implement the processing stages in industries.

Acetaminophen Overdose Enhances Early Cholangiocarcinoma in Opisthorchiasis Hamsters.

Opisthorchiasis which exerted by infection of Opisthorchis viverrini is strongly related to the incident of cholangiocarcinoma (CCA) in many Southeast Asian countries northeastern of Thailand. The O. viverrini infection is primarily caused by raw fish consumption, and repeated exposure to liver fluke. Meanwhile, acetaminophen is usually medicated to relieve pain in particularly people in northeast Thailand. OBJECTIVE: This study therefore aimed at investigating effects of acetaminophen on pathogenesis in hamsters for opisthorchiasis. METHODS: There were 4 groups of hamsters: i) uninfected hamster (N); ii) sole acetaminophen administration (N-Ac); iii) sole O. viverrini infection (OV); and iv) combination of O. viverrini infection and acetaminophen (OV-Ac) on pathology of hamsters for 1 month post infection. For analysis of histopathological changes through hematoxylin and eosin, Sirius red and immunohistostaining for Cytokeratin 19 (CK-19), Proliferating cell nuclear antigen (PCNA) and CA 19-9, serum's hamsters were used detected for liver function tests and tumor-related genes expression. RESULTS: After 1 month under these treatments, the OV-Ac showed significantly higher CCA risk, including inflammatory cells were aggregations around bile duct, new bile duct and fibrosis in subcapsular hepatic tissues, than other treatments. These pathological parameters were positively correlated with immunohistochemical staining derived from CK-19, PCNA and CA 19-9. In addition, OV-Ac had significantly higher liver function tests (ALT). CONCLUSION: Combined intake of liver fluke-contaminated raw fishes and acetaminophen rendered more severity of CCA than sole consumption of the contaminated raw fishes.

Antibiotic resistance in the pathogenic foodborne bacteria isolated from raw kebab and hamburger: phenotypic and genotypic study.

BACKGROUND: In recent years, interest in the consumption of ready-to-eat (RTE) food products has been increased in many countries. However, RTE products particularly those prepared by meat may be potential vehicles of antibiotic-resistance foodborne pathogens. Considering kebab and hamburger are the most popular RTE meat products in Iran, this study aimed to investigate the prevalence and antimicrobial resistance of common foodborne pathogens (Escherichia coli, Salmonella spp., Staphylococcus aureus, and Listeria monocytogenes) in raw kebab and hamburger samples collected from fast-food centers and restaurants. Therefore, total bacterial count (TBC), as well as the prevalence rates and antibiogram patterns of foodborne pathogens in the samples were investigated. Also, the presence of antibiotic-resistance genes (blaSHV, blaTEM, blaZ, and mecA) was studied in the isolates by PCR. RESULTS: The mean value of TBC in raw kebab and hamburger samples was 6.72 ± 0.68 log CFU/g and 6.64 ± 0.66 log CFU/g, respectively. E. coli had the highest prevalence rate among the investigated pathogenic bacteria in kebab (70%) and hamburger samples (48%). Salmonella spp., L. monocytogenes, and S. aureus were also recovered from 58, 50, and 36% of kebab samples, respectively. The contamination of hamburger samples was detected to S. aureus (22%), L. monocytogenes (22%), and Salmonella spp. (10%). In the antimicrobial susceptibility tests, all isolates exhibited high rates of antibiotic resistance, particularly against amoxicillin, penicillin, and cefalexin (79.66-100%). The blaTEM was the most common resistant gene in the isolates of E. coli (52.54%) and Salmonella spp. (44.11%). Fourteen isolates (23.72%) of E. coli and 10 isolates (29.41%) of Salmonella spp. were positive for blaSHV. Also, 16 isolates (55.17%) of S. aureus and 10 isolates (27.27%) of L. monocytogenes were positive for mecA gene. CONCLUSIONS: The findings of this study showed that raw kebab and hamburger are potential carriers of antibiotic-resistance pathogenic bacteria, which can be a serious threat to public health.

Raw Meat-Based Pet Feeding and Food Safety: Netnography Study of Pet Owner Comments and Review of Manufacturers' Information Provision.

ABSTRACT: Feeding raw meat to domestic pets is a popular practice. Because of the potential food safety implications associated with handling raw meat, concerns about pet owner health have increased. For this study, a netnographic content analysis approach was used to analyze posts from Pet Forums Community online archives related to raw meat-based feeding and food safety (n = 308). United Kingdom manufacturer and supplier Web sites (n = 33) were reviewed for provision of food safety information related to raw meat-based pet feeding. The data were analyzed using a qualitative thematic approach and quantitative methods. Pet owner forum comments indicated potential malpractices and indifference toward possible food safety hazards when preparing raw meat-based pet food. Many were risk aware but indicated confusion regarding appropriate food safety practices. Although emotional concern about the safety of pets and children was expressed, contracting a foodborne disease was not perceived as a personal risk. In addition, the review identified that most (61%) manufacturer and supplier Web sites failed to provide food safety instructions or warnings to pet owners regarding raw meat-based pet food. Information was inconsistent and varied across sources. The most comprehensive sources of information were provided by manufacturers approved by the United Kingdom Pet Food Manufacturers' Association. This is the first netnography study to explore pet owner online reports and the provision of food safety information related to raw pet feeding. Findings suggest pet owners may not fully appreciate the potential risks associated with raw meat-based pet feeding. Consequently, there is a need for credible and consistent strategies to inform pet owners about potential implications associated with feeding raw meat to pets, as well as about safe food handling practices.

Prevalence and associated risk factors of Shigella flexneri isolated from drinking water and retail raw foods in Peshawar, Pakistan.

This study was designed to investigate the prevalence and associated risk factors of Shigella flexneri isolated from drinking water and retail raw food samples in Peshawar, Pakistan. A total of 1,020 different samples were collected from various areas of Peshawar between January 2016 and May 2017, followed by identification of S. flexneri through biochemical, serological, and 16S rRNA gene sequencing. Potential risk factors associated with the development and spreading of S. flexneri infection were also investigated. Overall, 45 (4.41%) samples were positive for Shigella species. Among these samples, the predominant species was S. flexneri (n = 44) followed by S. boydii (n = 1). Interestingly, S. sonnei and S. dysenteriae isolates were not found in any sample. The isolation rate of S. flexneri in drinking water samples, market raw milk, and fruits/vegetables from Peshawar were 6.47%, 3.5%, and 2.9%, respectively. The phylogenetic reconstruction showed genetic diversity among three clades, as clades I and II have isolates of S. flexneri that were circulating within the drinking water, milk, fruits/vegetables, while clade III isolates were recovered from milk samples. Most of S. flexneri were detected in June to September. Potential risk factors of S. flexneri were water sources contaminated by toilet wastes (p = 0.04), surface water drainage (p = 0.0002), hospital wastes (p = 0.01), unhygienic handling (p < 0.05), and transportation of raw food (p = 0.04). In conclusion, S. flexneri isolates of closely related lineage originating from non-clinical samples might be associated with an increased human risk to shigellosis in Pakistan, as significant numbers of S. flexneri were observed in the drinking water and retail raw food samples. PRACTICAL APPLICATION: This study demonstrated the presence of S. flexneri in drinking water and retail raw food samples which seem to possess a serious threat to public health. Potential sources of food and water contamination should properly be monitored by public health authorities to reduce cases of shigellosis.

Rapid detection and enumeration of aerobic mesophiles in raw foods using dielectrophoresis.

The concept of dielectrophoresis (DEP), which involves the movement of neutral particles by induced polarization in nonuniform electric fields, has been exploited in various biological applications. However, only a few studies have investigated the use of DEP for detecting and enumerating microorganisms in foodstuffs. Therefore, we aimed to evaluate the accuracy and efficiency of a DEP-based method for enumerating viable bacteria in three raw foods: freshly cut lettuce, chicken breast, and minced pork. The DEP separation of bacterial cells was conducted at 20 V of output voltage and 6000 to 9000 kHZ of frequency with sample conductivity of 30-70 µS/cm. The accuracy and validity of the DEP method for enumerating viable bacteria were compared with those of the conventional culture method; no significant variation was observed. We found a high correlation between the data obtained using DEP and the conventional aerobic plate count culture method, with a high coefficient of determination (R2 > 0.90) regardless of the food product; the difference in cell count data between both methods was within 1.0 log CFU/mL. Moreover, we evaluated the efficiency of the DEP method for enumerating bacterial cells in chicken breasts subjected to either freezing or heat treatment. After thermal treatment at 55 °C and 60 °C, the viable cell counts determined via the DEP method were found to be lower than those obtained using the conventional culture method, which implies that the DEP method may not be suitable for the direct detection of injured cells. In addition to its high accuracy and efficiency, the DEP method enables the determination of viable cell counts within 30 min, compared to 48 h required for the conventional culture method. In conclusion, the DEP method may be a potential alternative tool for rapid determination of viable bacteria in a variety of foodstuffs.

A cluster of Shiga Toxin-producing Escherichia coli O157:H7 highlights raw pet food as an emerging potential source of infection in humans.

In August 2017, a cluster of four persons infected with genetically related strains of Shiga toxin-producing Escherichia coli (STEC) O157:H7 was identified. These strains possessed the Shiga toxin (stx) subtype stx2a, a toxin type known to be associated with severe clinical outcome. One person died after developing haemolytic uraemic syndrome. Interviews with cases revealed that three of the cases had been exposed to dogs fed on a raw meat-based diet (RMBD), specifically tripe. In two cases, the tripe had been purchased from the same supplier. Sampling and microbiological screening of raw pet food was undertaken and indicated the presence of STEC in the products. STEC was isolated from one sample of raw tripe but was different from the strain causing illness in humans. Nevertheless, the detection of STEC in the tripe provided evidence that raw pet food was a potential source of human STEC infection during this outbreak. This adds to the evidence of raw pet food as a risk factor for zoonotic transmission of gastrointestinal pathogens, which is widely accepted for Salmonella, Listeria and Campylobacter spp. Feeding RMBD to companion animals has recently increased in popularity due to the belief that they provide health benefits to animals. Although still rare, an increase in STEC cases reporting exposure to RMBDs was detected in 2017. There has also been an increased frequency of raw pet food incidents in 2017, suggesting an increasing trend in potential risk to humans from raw pet food. Recommendations to reduce the risk of infection included improved awareness of risk and promotion of good hygiene practices among the public when handling raw pet food.

Recombinant FimH, a fimbrial tip adhesin of Vibrio parahaemolyticus, elicits mixed T helper cell response and confers protection against Vibrio parahaemolyticus challenge in murine model.

Vibrio parahaemolyticus causes vibriosis in wide range of marine organisms, and is responsible for food borne illnesses in humans through consumption of contaminated uncooked/partially cooked seafood. Continued and widespread antibiotics usage to increase the productivity has led to antibiotics resistance development. This has necessitated the need to develop alternative methods to control its infection. Use of safe and effective vaccines against the virulence factors not only protects from infection, it also minimizes antibiotic usage. The colonization of V. parahaemolyticus in the host and disease development requires several adhesins present on the cell surface, and thereby make them attractive vaccine candidates. V. parahaemolyticus produces extracellular type 1 fimbriae that have been shown to play a role in adhesion, biofilm formation and virulence. FimH is one of the minor components of the type 1 fimbriae occurring on its very tip. Being present on the cell surface, it is highly immunogenic, and can be targeted as a potential vaccine candidate. The present study describes the immunogenic and vaccine potential of recombinant V. parahaemolyticus FimH (rVpFimH) expressed in E. coli. Immunization of BALB/c mice with the rVpFimH elicited a strong mixed immune response, T-cell memory (evidenced by antibody isotyping, cytokine profiling and T-cell proliferation assay), and agglutination positive antibodies. FACS analysis and immunogold labeling showed that the polyclonal anti-rVpFimH antibodies were able to recognize the FimH on V. parahaemolyticus cells. In vivo challenge of the rVpFimH-immunized mice with 2×LD50 dose of live bacteria showed one hundred percent survival. Thus, our findings clearly demonstrate the potential of FimH as an effective vaccine candidate against V. parahaemolyticus.

Microbial quality of raw cow milk and its predictors along the dairy value chain in Southwest Ethiopia.

Raw milk may contain pathogenic microorganism that can seriously affect the health of consumers. In Southwest Ethiopia, raw cow milk is consumed more than the processed products, but its microbiological quality and its predictors are not studied well. The aim of this study was to determine the microbial quality of raw cow milk and its predictors along the dairy value chain in Southwest Ethiopia. A total of 150 milk and 300 environmental samples were collected randomly from dairy farms, milk distribution centers, and retailer outlets for microbiological analysis using standard protocols. One milk handler from each milk production or distribution stage was also interviewed to assess the knowledge, attitude, and practices regarding milk handling. Descriptive statistics and multiple linear regression models were used to summarize the data and to identify predictors of milk microbial quality, respectively. As the milk transported from dairy farm to milk retailer outlet, the mean total bacterial count has increased from 5.0 ±â€¯0.3 to 7.2 ±â€¯0.1 log CFU/ml respectively. The mean coliform count of the milk sample was 4.4 ±â€¯0.4 log CFU/ml at the dairy farm and 7.0 ±â€¯0.2 log CFU/ml at milk selling points, indicating the deterioration of milk quality along the dairy value chain. All of the analyzed water samples were positive for fecal coliform bacteria. The highest coliform bacteria were reported from milk storage equipment found at milk retailer outlet with the count of 4.8 ±â€¯0.5 log CFU/ml. Educational status and attitude of milk handlers and the quality of water used to wash milk equipment and hands of milk handlers were the major factors affecting the microbial quality of raw cow milk. The findings of this study revealed that the microbial quality of raw milk in the study area was poor. Hence, improving the attitude and educational status of milk handlers, and the quality of water is an important step to enhance milk quality and consequently to prevent milk borne diseases.