Raw Unpasteurized Milk Research Articles

The prevalence, antimicrobial resistance and molecular genetic characteristics of fosfomycin-resistant Klebsiella pneumoniae isolated from raw milk

This investigation aims to explore the molecular epidemiological characteristics of the fosfomycin resistance gene, fosA5, in unpasteurized raw milk. A total of 385 unpasteurized raw milk samples were collected from dairy farms in Jiangsu province, China. From these samples, 102 strains of Klebsiella pneumoniae were isolated, with 9 of them testing positive for the presence of the fosA5 gene. Antimicrobial susceptibility testing revealed that K. pneumoniae carrying fosA5 exhibited high-level resistance to fosfomycin but also to other tested antibiotics, indicating multidrug resistance. The S1-PFGE and conjugation results indicated that the fosA5 gene was located on the chromosome. Furthermore, the analysis of PFGE patterns, MLST, and WGS elucidated the genetic diversity among K. pneumoniae strains harboring the fosA5 gene, indicating both clonal and horizontal transmission. This study highlights the prevalence and characteristics of the fosA5 fosfomycin resistance gene in large-scale dairy farms in Jiangsu province. Given its potential public health risks, these findings warrant significant attention.

Digestive Pathologies Caused by E. coli Bacteria in Non-Treated Dairy Products

The purpose of this review is to summarize current possibility of contamination of milk and dairy products and human pathologies as well, that may occur if these products are not processed properly. Scientific literature was consulted to examine the methods of contamination of milk and dairy products. In raw or pasteurized milk stored at 5 °C was not reported that it can grow E. coli, however can grow slowly at higher temperatures. E. coli is considered a high-risk potential pathogen in cheese, due to its unusual ability to tolerate low pH values for long periods and it’s linked to unpasteurized raw milk.

Seroprevalence and molecular detection of brucellosis among Pakistani women with spontaneous abortion.

Human brucellosis is a neglected disease transmitted to humans from animals such as cattle, goats, dogs, and swine. The causative agents are bacteria of the genus Brucella, intracellular pathogens usually confined to the reproductive organs of their animal hosts causing sterility and abortions. The objective of the study was to determine the seroprevalence of brucellosis among women with spontaneous abortions (SAW) and compare this seroprevalence with that of healthy pregnant women (HPW). The case-control study was designed to determine the seroprevalence and molecular detection of brucellosis in women who suffered from spontaneous abortion and healthy pregnant women of the Haripur District of Pakistan. A total of 770 blood samples (n = 385 for each group) were collected from 9 public and 11 private hospitals in Haripur District from December 2021-March 2023. Data on demographic features, epidemiological variables, and risk factors were collected from each participant by structured questionnaires. Initial screening for brucellosis was performed by Rose Bengal Plate Test followed by qRT-PCR for molecular detection of the genus-specific BCSP-31 gene of Brucella. The study showed that anti-Brucella antibodies were more found in SAW 23.63% (91/385) than in HPW 1.29% (5/385). Brucella specific DNA was amplified in 89.01% (81/91) seropositive samples of SAW. Demographic features and risk factors such as age, urbanicity, socioeconomic status, education, occupation, and animal contact were found significantly associated with brucellosis (p ≤ 0.05). Consumption of unpasteurized raw milk (OR = 18.28, 95%CI: 8.16-40.94) was found highly concomitant with seroprevalence. This study reports the first evidence of involvement of brucellosis in spontaneous abortions in women of Pakistan. The study can be used to develop strategies for risk management during pregnancy, to raise awareness for brucellosis, and develop control programs.

Real-time and visual detection of viable Salmonella in milk from remote pasture via IMS-LAMP-NALFS

Salmonella is a prevalent foodborne pathogen commonly found in dairy products. Controlling the presence of Salmonella during dairy product production is of utmost concern due to the potential for contamination at any stage of the production chain, particularly at the source. Moreover, the unique location of pastures and limited testing resources further highlight the necessity of establishing a rapid and precise detection method to ensure milk quality control and enhance overall farm safety. To address these issues, in this study, we constructed a method as well as a portable toolbox for rapid on-site detection of Salmonella. The method involves immunomagnetic separation (IMS) using streptavidin-coupled magnetic nanoparticles and biotinylated antibodies. The reaction conditions for magnetic capture and enrichment of Salmonella were optimized to efficiently remove matrix interference from milk and enhance detection sensitivity. Additionally, an improved propidium monoazide (PMAxx) treatment was employed to eliminate false positive results caused by dead bacteria. This treatment was combined with loop-mediated isothermal amplification (LAMP) and nucleic acid lateral flow strip (NALFS) (LAMP-NALFS) for on-site detection of viable bacteria in milk. The resulting IMS-LAMP-NALFA assay achieved rapid and sensitive detection of viable Salmonella, with a detection sensitivity of 1 × 102 CFU/mL in milk samples, completed within 90 min under optimal conditions. Based on this achievement, a portable toolbox was designed for easy transportation and on-site testing in pastures. It is equally applicable for rapid detection of viable Salmonella contamination in food products in resource-limited areas such as pastures, where the observed detection rate of Salmonella in unpasteurized raw milk was 7.93 % (28 out of 353 samples tested). Consequently, the IMS-LAMP-NALFA method and the associated portable toolbox represent highly effective means of enhancing the detection of viable Salmonella in dairy products.

Status and Public Health Significance of Mycobacterium bovis in Ethiopia

Infectious diseases, accounting for 30-50% of the total annual losses, remain a major impediment to Ethiopia’s livestock economy. Cattle, other domesticated animals, and some free-range or captive wildlife species are all susceptible to the infectious disease known as bovine tuberculosis, which is caused by the Mycobacterium bovis (M. bovis). It is typically distinguished by the development of tubercle-like nodular granulomas. The bovine tuberculosis-diseased animal loses 10-25% of their productive efficiency; direct losses due to the infection become evident by a decrease in 10-18% milk and a 15% reduction in meat production. During the final stages of tuberculosis, severe emaciation and acute respiratory distress might happen. It affects animal production, but it also significantly affects public health. Human infection due to M. bovis is thought to be mainly through the drinking of contaminated or unpasteurized raw milk and undercooked meat. It is estimated that M. bovis causes 10-15% of human cases of tuberculosis in countries. This indicated that tuberculosis in both humans and animals is endemic in Ethiopia. Reporting the status of tuberculosis in animals and humans is so dynamic.

Factors influencing somatic cell counts and bacterial contamination in unpasteurized milk obtained from water buffalo in Bangladesh

Little has been published on the factors influencing the safety and quality of milk derived from water buffalo in Bangladesh. This study aims to describe the milk hygiene parameters and milk chain characteristics of unpasteurized raw milk sold to consumers in order to improve milk hygiene. A quantitative study design evaluated somatic cell counts, total bacterial counts, and specific gram-negative (Enterobacteria) and gram-positive (staphylococci) pathogens in 377 aseptically collected milk samples. Samples were collected at multiple nodes along the buffalo milk value chain: 122 bulk tank milk samples were collected at the farm level, 109 milk samples at the middlemen level, and 111 milk samples at the milk collection centers. In addition, 35 samples were taken from various milk products at the retail level. It was found that progressively increasing somatic cell counts and bacterial counts, including potential pathogens, occurred along the milk chain. A seasonal increase in spring was found, varying based on the farming system (semi-intensive versus intensive). Other factors included water purity and cleanliness of containers, mixing buffalo and cow’s milk, and the location of the water buffalo milk producer (coastal or river basin). This study demonstrated how improving udder health and milk hygiene along the water buffalo milk value chain would increase the safety and quality of water buffalo milk in the study area.

MRSA in bulk tank milk of dairy herds in Germany - changes over time.

Methicillin-resistant Staphylococcus aureus (MRSA) have repeatedly been isolated from dairy herds. It was the purpose of this study to compare the results of 3 subsequent national scale cross-sectional investigations in dairy herds in Germany on the prevalence of MRSA in bulk tank milk and the characteristics of the isolates. The investigations were carried out in 2010, 2014 and 2019, respectively. MRSA were isolated from 25 ml of bulk tank milk using a double selective enrichment protocol. Samples were distributed across the country according to the regional dairy cattle population. The prevalence of MRSA in bulk tank milk samples was lower in 2010 than in 2014 and tended to decrease until 2019. Prevalence was higher in samples from conventional than from organic herds and increased with herd size. Most isolates (75/78) were assigned to the clonal complex 398 and the spa-types t011 and t034. Resistance of the isolates to other antimicrobials than beta-lactams decreased over time. MRSA remain present in the German dairy population and are found more frequently in larger vs. smaller herds and in conventional vs. organic herds. MRSA should be considered in biosecurity protocols and with respect to occupational health of farm staff. Presence of MRSA in raw milk supports the recommendation not to drink unpasteurized raw milk.

SCREENING OF ANTIBIOTIC RESISTANCE AND VIRULENCE GENES OF ENTEROCOCCUS SPP. STRAINS ISOLATED FROM URFA CHEESE

Enterococcus faecium, E. durans and E. faecalis species were isolated and identified from traditional Urfa cheese samples which were produced from unpasteurized raw milk. The enterococcal load of the cheese samples was between 4.4-5.6 log cfu g−1. High-level tetracycline, streptomycin, erythromycin, gentamycin, and penicillin resistance was determined in the enterococcal isolates. Multiple antibiotic resistance was also determined in E. faecalis (20.4%) and E. faecium (16.3%) strains. 36.7% of the enterococcal isolates were greater than 0.2 MAR index ratio in this study. The gelE and agg2 genes were found in 40 (81.63%) of the enterococcal isolates, whereas the vanB gene was found in 3 (6.12%) of the enterococcal isolates. The results indicate that the consumption of Urfa cheese, which is produced using raw milk, may have public health risk because of its antibiotic resistance characteristics and virulence genes of enterococcal biota.

Prevalence and Phylodiversity of ESBL-Producing Coliforms Isolated from Ruminant Mastitis in Nigeria

Abstract The public health threat posed by Extended-spectrum beta-lactamase producing E. coli (ESBL-EC) in food animal production systems has attracted global attention. Data on the prevalence, diversity and genetic characteristics of ESBL-producing coliforms are key to advocacy on promoting responsible antimicrobial stewardship and proper planning of control strategies. The coliforms were isolated from 1052 milk samples of 160 cows, 103 ewes and 103 does with mastitis in Plateau State, Nigeria and analysed for ESBL production by phenotypic, biochemical, antimicrobial sensitivity and genetic characterization. The percentage of occurrence of clinical mastitis in cows, ewes, and does were 0.2 %, 0 %, 1.5 % respectively, while the percentage occur-rence of subclinical mastitis in ruminants were 18.1 %, 28.2 % and 38.3 % respectively. From the 677 isolates, 31.3 % (n = 212) were ESBL producing coliforms, with a prevalence of 48.6 %, 18.4 %, 12.7 %, 8.9 %, 5.7 %, 3.8 % and 1.9 % for E. coli, K. pneumoniae, C. freundii, K. aerogenes, S. marcescens, K. oxytoca and E. cloacae, respectively. The genetic characterization revealed a higher prevalence of bla CTX-M than bla TEM in the samples analysed (24.39 % vs. 12.19 %). High pairwise identity was observed among the bla CTX-M and bla TEM gene sequences obtained in this study, but they displayed high phylodiversity with sequences from ruminants and humans from other climes. The bla SHV gene was not detected. Multidrug resistances especially to the commonly used antimicrobials; ofloxacin, gentamycin and streptomycin in veterinary practice in Nigeria were observed. This has public health implications considering the fact that consumption of raw unpasteurized milk is a common practice in some cultures in Nigeria. Such practise will facilitate the transfer of multidrug resistant coliforms to humans resulting in the complications of treatment outcomes. To the best of our knowledge this is the first genetic characterization of ESBL-producing agents from ruminant mastitis in Nigeria.

COMPARATIVE ANALYSIS OF DIFFERENT PATHOGENIC MICROBE COLLECTED FROM MAMMALIAN MILK

Milk comprises proteins, fats, minerals, carbohydrates, multiple vitamins, and various special substances dispersed in water. Different types of pathogenic microbes present in unpasteurized raw milk, cause gastrointestinal tract infections and food-borne infections. The current study was based on evaluating pathogenic microbe’s frequencies among the mammal (buffalo, cow, camel, goat and sheep) milk and assessing their nutritional parameters along with the screening of antibiogram of these pathogens in the district Peshawar, Pakistan. A total of 25 milk samples (05 from each mammals) were collected. Among the analyzed milk samples, S. aureus and Shigella spp. were observed n= 04 (16%), while E. coli, Salmonella spp., and P. aeruginosa were n= 03 (12%). The S. aureus was found highly resistant (100%) to Meropenem, Gentamicin, Amoxicillin, Aztreonam, and Doxycycline, while found highly sensitive (50%) to Amikacin, and Levofloxacin. E. coli showed high resistant (100%) to Meropenem, Ciprofloxacin, Amoxicillin, Ampicillin, and Doxycycline, while found to be highly sensitive (50%) to Levofloxacin, and Cefotaxime. The Salmonella spp. was found highly resistant (100%) to Amikacin, Meropenem, Amoxicillin, Cefotaxime, Ampicillin, Aztreonam, and Doxycycline, while found highly sensitive (50%) to Ciprofloxacin, and Gentamicin. The Shigella spp. was found highly resistant (100%) to Meropenem, Amoxicillin, Ampicillin, and Doxycycline, while found highly sensitive (25%) to Amikacin, Levofloxacin, Ciprofloxacin, Gentamicin, Cefotaxime, and Aztreonam. The P. aeruginosa was highly resistant (100%) to Amikacin, Levofloxacin, Ciprofloxacin, Gentamicin, Cefotaxime, Amoxicillin, Aztreonam, Doxycycline, while found highly sensitive (25%) to Meropenem, and Ampicillin. The high ESBL-producing bacterial pathogen was found E. coli (66.66%), followed by Shigella spp. (50%), Salmonella spp., and P. aeruginosa (33.33%), respectively. The nutritional analysis of mammal milk samples significantly (p < 0.05) with different parameters. Among these parameters, the fat Mean ± SD (3.36 ± 0.11), protein content Mean ± SD (2.91 ± 0.04), lactose content Mean ± SD (4.31 ± 0.10), and total solid content Mean ± SD (11.61 ± 0.27) were observed. In conclusion, of the current study, pathogenic microbes are present in unpasteurized milk, which shows high resistance to various antibiotics, and the nutritional values of milk were also low.

Whole-Genome Sequencing of Gram-Negative Bacteria Isolated From Bovine Mastitis and Raw Milk: The First Emergence of Colistin mcr-10 and Fosfomycin fosA5 Resistance Genes in Klebsiella pneumoniae in Middle East.

Antimicrobial resistance is a major concern in the dairy industry. This study investigated the prevalence, antimicrobial resistance phenotypes, and genome sequencing of Gram-negative bacteria isolated from clinical (n = 350) and subclinical (n = 95) bovine mastitis, and raw unpasteurized milk (n = 125). Klebsiella pneumoniae, Aeromonas hydrophila, Enterobacter cloacae (100% each), Escherichia coli (87.78%), and Proteus mirabilis (69.7%) were the most prevalent multidrug-resistant (MDR) species. Extensive drug-resistance (XDR) phenotype was found in P. mirabilis (30.30%) and E. coli (3.33%) isolates. Ten isolates (four E. coli, three Klebsiella species and three P. mirabilis) that displayed the highest multiple antibiotic resistance (MAR) indices (0.54–0.83), were exposed to whole-genome sequencing (WGS). Two multilocus sequence types (MLST): ST2165 and ST7624 were identified among the sequenced E. coli isolates. Three E. coli isolates (two from clinical mastitis and one from raw milk) belonging to ST2165 showed similar profile of plasmid replicon types: IncFIA, IncFIB, IncFII, and IncQ1 with an exception to an isolate that contained IncR, whereas E. coli ST7624 showed a different plasmid profile including IncHI2, IncHI2A, IncI1α, and IncFII replicon types. ResFinder findings revealed the presence of plasmid-mediated colistin mcr-10 and fosfomycin fosA5 resistance genes in a K. pneumoniae (K1) isolate from bovine milk. Sequence analysis of the reconstructed mcr-10 plasmid from WGS of K1 isolate, showed that mcr-10 gene was bracketed by xerC and insertion sequence IS26 on an IncFIB plasmid. Phylogenetic analysis revealed that K1 isolate existed in a clade including mcr-10-harboring isolates from human and environment with different STs and countries [United Kingdom (ST788), Australia (ST323), Malawi (ST2144), Myanmar (ST705), and Laos (ST2355)]. This study reports the first emergence of K. pneumoniae co-harboring mcr-10 and fosA5 genes from bovine milk in the Middle East, which constitutes a public health threat and heralds the penetration of the last-resort antibiotics. Hence, prudent use of antibiotics in both humans and animals and antimicrobial surveillance plans are urgently required.

Virulence Determinants and Plasmid-Mediated Colistin Resistance mcr Genes in Gram-Negative Bacteria Isolated From Bovine Milk.

A major increase of bacterial resistance to colistin, a last-resort treatment for severe infections, was observed globally. Using colistin in livestock rearing is believed to be the ground of mobilized colistin resistance (mcr) gene circulation and is of crucial concern to public health. This study aimed to determine the frequency and virulence characteristics of colistin-resistant Gram-negative bacteria from the milk of mastitic cows and raw unpasteurized milk in Egypt. One hundred and seventeen strains belonging to Enterobacteriaceae (n = 90), Pseudomonas aeruginosa (n = 10), and Aeromonas hydrophila (n = 17) were screened for colistin resistance by antimicrobial susceptibility testing. The genetic characteristics of colistin-resistant strains were investigated for mcr-1–9 genes, phylogenetic groups, and virulence genes. Moreover, we evaluated four commonly used biocides in dairy farms for teat disinfection toward colistin-resistant strains. Multidrug-resistant (MDR) and extensive drug-resistant (XDR) phenotypes were detected in 82.91% (97/117) and 3.42% (4/117) of the isolates, respectively. Of the 117 tested isolates, 61 (52.14%) were colistin resistant (MIC >2 mg/L), distributed as 24/70 (34.29%) from clinical mastitis, 10/11 (90.91%) from subclinical mastitis, and 27/36 (75%) from raw milk. Of these 61 colistin-resistant isolates, 47 (19 from clinical mastitis, 8 from subclinical mastitis, and 20 from raw milk) harbored plasmid-borne mcr genes. The mcr-1 gene was identified in 31.91%, mcr-2 in 29.79%, mcr-3 in 34.04%, and each of mcr-4 and mcr-7 in 2.13% of the colistin-resistant isolates. Among these isolates, 42.55% (20/47) were E. coli, 21.28% (10/47) A. hydrophila, 19.12% (9/47) K. pneumoniae, and 17.02% (8/47) P. aeruginosa. This is the first report of mcr-3 and mcr-7 in P. aeruginosa. Conjugation experiments using the broth-mating technique showed successful transfer of colistin resistance to E. coli J53-recipient strain. Different combinations of virulence genes were observed among colistin-resistant isolates with almost all isolates harboring genes. Hydrogen peroxide has the best efficiency against all bacterial isolates even at a low concentration (10%). In conclusion, the dissemination of mobile colistin resistance mcr gene and its variants between MDR- and XDR-virulent Gram-negative isolates from dairy cattle confirms the spread of mcr genes at all levels; animals, humans, and environmental, and heralds the penetration of the last-resort antimicrobial against MDR bacteria. Consequently, a decision to ban colistin in food animals is urgently required to fight XDR and MDR bacteria.

Molecular Detection of Coxiella burnetii in Unstandardized Minas Artisanal Cheese Marketed in Southeastern Brazil

Q fever is a zoonotic disease caused by Coxiella burnetii. The causative pathogen has been detected in dairy products, which raises the possibility of consumption of dairy products as a possible route of transmission; however, this has not been adequately described. Therefore, the aim of the present study was to investigate the presence of C. burnetii in 87 samples of artisanal cheeses made from unpasteurized raw milk sold in Minas Gerais, southeastern Brazil. The cheese samples were analyzed using quantitative real-time PCR (qPCR), and the products from positive samples were sequenced. In addition, 150 people were interviewed to outline the profile of consumers and the consumption of non-inspected dairy products. The results showed that 4.6% (4/87) of the samples were positive for C. burnetii, with 99.84% identity with the reference 16S gene. Responses from consumer interviews indicate that 95.3% (143/150) of respondents consume milk and dairy products; 50% (75/150) consume fresh Minas artisanal cheese and milk at least once a day; 64.58% (93/144) do not check the inspection stamps on the packaging at the time of purchase; and 81.56% (122/146) do not know the meaning of these stamps. Thus, the presence of C. burnetii DNA in the analyzed cheeses and the consumption of unstandardized dairy products raise an alert for the risk of transmission of Q fever in the population that consumes them.

Milk from healthy or infected cattle as a source of multi-drug resistant, AmpC β-lactamase-producing Escherichia coli

Drinking raw unpasteurized milk or products made from raw milk is a recent trend observed among the people due to their belief in enhancing nutritional properties and better taste. It poses a great risk of microbial transmission along with antibiotic resistance gene pool (extended spectrum beta-lactamase or ampC) among the consumers. The study was conducted to detect the occurrence of AmpC producing E. coli in raw milk samples collected from healthy and mastitic cattle in West Bengal. In total, 450 cow milk samples were collected from all the six agro-climatic zones of West Bengal. The samples were collected from healthy as well as infected cattle suffering with clinical or sub-clinical mastitis, irrespective of age and breed. The collected milk samples were tested with MAST INDEX reagent for the detection of sub-clinical mastitis. E. coli isolates obtained from cattle milk samples (n=450) were subjected to cefoxitin-cloxacillin double disc synergy test and PCR based confirmation of blaAmpC, class I integron and CITM genes. All the ACBL producing E. coli isolates were subjected for their resistance to other antimicrobials. All the levofloxacin and Co-Trimoxazole resistant ACBL-producing isolates were studied for PMQR, sul1, sul2, sul3 genes. E. coli was isolated from 205 collected milk samples (205/450, 45.6%). The phenotypical test detected 231/383 isolates (60.3%) were AMPC producers which were also found to possess blaAmpC. Comparison of ACBL-producing E. coli occurrence in cattle with mastitis, sub-clinical mastitis and healthy cattle revealed significantly higher occurrence rate in infected cattle than the healthy ones (p<0.05). Phenotypical antibiotic resistance pattern of the isolates showed resistance against penicillin-G (100%), cefoxitin (100%), ampicillin (42%), co-trimoxazole (13.5%) and levofloxacin (5.2%). Out of 30 co-trimoxazole resistant isolates identified, 25 (83%) and seven (30.4%) isolates possessed sul1 and sul2 gene, respectively, whereas, four (13.3%) isolates possessed both sul1 and sul2 genes. Three levofloxacin resistant isolates possessed qnrS gene. This is the first systemic study with substantial numbers of milk samples from healthy and infected cattle depicting occurrence of ACBL-producing E. coli and linkage present in the isolates between the genes encoding ACBL and quinolone or sulphonamide resistance.

Advanced Methods for Isolating from and Confirming Campylobacter spp. in Milk and Dairy Products: Review

Campylobacter spp. are a type of microaerophilic bacteria that cause human foodborne illnesses worldwide. Among the various types of Campylobacter spp., Campylobacter jejuni and Campylobacter coli account for 90% of foodborne campylobacteriosis. Generally, poultry meats are known to be a primary cause of campylobacteriosis; however, several other types of foods have also been reported to cause campylobacteriosis. Particularly, raw milk has been directly linked to Campylobacter infections among many foodborne illnesses, and cases of campylobacteriosis caused because of the ingestion of unpasteurized raw milk have been recorded worldwide. This review reports (1) general information, history, and nomenclature of Campylobacter spp., (2) epidemiology of Campylobacter spp., (3) detection of Campylobacter spp. from foods including milk and dairy products, and (4) review of methods for controlling the growth Campylobacter spp.

Fate and survival of Listeria monocytogenes and Escherichia coli O157:H7 during ripening of cheddar cheeses manufactured from unpasteurized raw milk

Cheese products may serve as vehicles for transmission of foodborne pathogens, causing disease outbreaks, particularly when unpasteurized raw milk is used for cheese production. Therefore, the adequacy of a 60-day aging period for raw milk Cheddar cheese to eliminate Escherichia coliO157:H7 and Listeria monocytogenes was evaluated. The changes in cheese components and characteristics of pH, Aw, coliform, and LAB were examined during the ripening process. Cheddar cheese was manufactured from raw milk artificially spiked with 1, 3, or 5 log CFU/mL L. monocytogenes or E. coliO157:H7. When compared with the pasteurized samples, the unpasteurized samples exhibited slightly lower pH values and similar aw values. No coliform were observed in the pasteurized cheese, whereas over 5-log CFU/g of coliform were found in the cheese made from unpasteurized milk during the early phases of ripening. However, coliform were completely eliminated from the cheese made from unpasteurized milk within five weeks. In contrast, while the level of LAB was initially 2-log lower in the cheese made from pasteurized milk than that derived with unpasteurized milk, the number of LAB between the two cheese preparations equalized within five weeks. Although L. monocytogenes numbers decreased significantly to below the detection limit (1-log CFU/g) during ripening, viable cells were isolated from all the inocula. E. coliO157:H7 cells also significantly decreased to below the detection limit during ripening at all the inoculation levels. However, viable cells were still detected. Thus, a 60-day ripening process alone may not be sufficient for elimination of contaminating pathogens.

Detection of Aeromonas hydrophila in Raw Milk and Soft Cheese in Baghdad City

The frequency and distribution pattern of Aeromonas hydrophilain cow’s raw milk and soft cheese were investigated in Abu-Ghraib, Al-Fudhaliyah and Al-Sadrya regions in Baghdad. A total of one hundred twenty pooled samples were collected during February until June (2019),in which sixty samples of raw milk pooled from milk containers, twenty from each region (four samples collected monthly per region) and sixty samples of fresh soft cheese made from raw unpasteurized milk were collected from different supermarkets in Baghdad as same as proportion above. Modified procedures for isolation and identification were dependent in which, modified tryptone soya yeast extract sheep blood agar supplemented with ampicillin used. Gram stain and oxidase reaction aid in segregation. Modified Congo red assay used to detect biofilm and Kirby-Bauer disc diffusion method was used for determination sensitivity of isolates toampicillin (AM 10µg),cephalexin (CLX, 30µg), azithromycin (AZM, 15µ) andvancomycin (VA 30µg). The results confirmed recovery of eleventh isolates (9.16 %) from one hundred twenty pooled samples, in which four isolates (3.33 %) from raw milk samples; Two (1.66 %) from Abu-Ghraib and one(0.83 %) from each other regions. Seven isolates (5.83 %) were detected from fresh soft cheese samples in which, three (2.5 %) from Abu-Ghraib and two (1.66 %) from each other regions. All isolates were Gram-negative rod- shaped, oxidase positive and biofilm producers. Four isolates (36.36 %) were resistant to selected antibiotics in which, two (18.18 %) from Abu-Ghraib: One (9.09 %) from raw milk and soft cheese; and one (9.09 %) in each other regions in fresh soft cheese only. In conclusions. Milk and cheese production in Abu-Ghraib, Al-Fudhaliyah and Al-Sadrya regions in Baghdad are encountered by the problems of bacterial contamination and presence of multidrug resist strain of A. hydrophila which considered a risk to public health, may be due both insufficient and misuse treatment with antibiotics or in somewhat due to bad quality and/or poor hygiene of processing involved in milk production. Thus recommended monitoring these products for better hygienic status

Acute and chronic brucellosis eleven-year audit from a tertiary hospital in Armenia.

The incidence of brucellosis in Armenia in 2010 was twice as high as in other countries of the Caucasian region and has almost doubled over the last three decades. This study aimed to investigate factors associated with acute or chronic forms of presentation of human brucellosis. Retrospective study using data from medical records of 455 patients hospitalized for the first time at the Nork Republican Infectious Disease Referral Hospital in Yerevan, Armenia between the years 2006 and 2016. We undertook descriptive analysis of cases, compared acute and chronic cases, and identified factors associated with acute and chronic cases using regression. The majority of brucellosis cases had acute case presentation (73.0%), were males (70.3%), between the ages of 20-60 years (66.2%) and unemployed (89.9%). About two-thirds of cases reported a history of consumption of raw unpasteurized milk. The multivariate analysis revealed that factors associated with the form of brucellosis were age, symptom duration preadmission, fever, antibody titer, and hospitalization outcomes. This study revealed that brucellosis is unevenly distributed across different age groups, as well as regions of Armenia. Affected individuals did not seek medical attention after the onset of the symptoms for about 2 months. Therefore, the targeted educational campaigns could be of crucial importance to prevent the disease in humans, contribute to its early diagnosis and treatment.

Burden of human brucellosis and risk factors in animal-human interface: A case study in Mikumi-Selous ecosystem, Morogoro Tanzania

Fever causing zoonoses such as brucellosis are common in sub-Saharan countries and rarely routinely diagnosed in patients with febrile conditions. The aim of this cross sectional study was to establish the burden and risk factors of human brucellosis in animal-human interface areas in Mikumi-Selous ecosystem, Tanzania. The study was undertaken from January 2011 to February 2012. Brucellosis screening was carried out by Rose Bengal Plate test (RBPT) in serum from fever (n=1509) and none fever (n=298) patients attending at selected health facilities. The overall sero-prevalence of brucellosis in human was 20.5% (95% CI: 18.7-22.5, n=1807). A highly significant ( P = 0.0001) infection rate was found in fever group 23.9% (n = 1509) than individuals in non-fever group (3.7%). Mvomero district had higher prevalence ( P = 0.0001, OR = 1.42) (36.1%) than other districts. Questionnaires were administered to 369 brucellosis screened respondents in their households. It was found that contact with cattle manure, milking work, and contact with placenta during assisted parturition and home slaughter were the significant risk factors for transmission of brucellosis by direct contact. However, consumption of raw unpasteurized milk, undercooked or raw meat and raw blood constituted a major threat of brucellosis to the community in the study area. Brucellosis is prevalent in human distributed in all districts posing a health threat to pastoral and agro-pastoral communities. Participatory intervention through One-Health approach coupled with effective mass vaccination program in animals is vital for control and eradication of the disease. Keywords: Brucellosis, humans, animals, Mikumi-Selous Ecosystem, risk factors

Detection of Salmonella Spp. in milk samples of selected regions of Diyala city

The aim of this study was to identify milk contamination that produced from dairy cattle with salmonella organisms in different regions of Diyala city. Fresh (unpasteurized) and heat treated (pasteurized) milk samples (n=10) were collected randomly from non-similar five regions of Diyala city that well-known with dairy cattle production particularly local shops and cattle shepherd markets, samples were send to the laboratory for some microbial testing. Special selective media were prepared and cultured with pasteurized and raw unpasteurized milk, the results refered that there were infected milk samples out of 10 fresh unpasteurized milk as the following percentage, 10% Rashdiah, 20% Bani saad, 80% Ghalbiah, 50% Khalis and 20 % Shiftah, while the pasteurized milk samples displayed contamination out of 10 samples as the previous (20% Rashdiah and 10% Ghalbiah), other pasteurized samples regions showed negative results for salmonella in regions of Bani saad, Khalis and Shiftah. We concluded that both heat treated and fresh milk may be unsafe for humankind ingesting if produced under unsanitary conditions or insufficient pasteurization.