scholarly journals Isolation and Identification of Pathogenic Staphylococci and E. Coli From Raw Bovine Milk Collected From Milk Cooperative Centers in Hawassa, Southern Ethiopia

Author(s):  
Alemayehu Yenealem

Abstract Background: Pathogenic microorganisms commonly isolated from milk and milk products pose a serious threat to human health. Escherichia coli and Staphylococci are the major pathogens isolated from milk. The objective of this study was to isolate and identify pathogenic Staphylococci and E.coli from raw bovine milk collected from milk cooperative centers found in Hawassa City, southern Ethiopia. Result: The overall prevalence of pathogenic species of Staphylococci and E. coli was found to be35.16 % and 8.59 % (n=384), respectively. From the total 384 raw milk samples examined, 1.56 %was found to be positive for both pathogenic species of Staphylococci and E. coli. The prevalence of pathogenic species of Staphylococci was found to be 33.33 %, 36.05 % and 39.21 % milk collected from Arsi Negele, Hawassa, and Shashemene, respectively. Whereas the prevalence of E. coli was found to be 9.68 %, 8.84 % and 3.92 %) milk collected from Arsi Negele, Hawassa and Shashemene, respectively. The study results showed a relatively higher prevalence of pathogenic species of staphylococci in raw milk collected from Shashemene than raw milk collected from Arsi Negele andHawassa. However, the difference was statistically insignificant (p > 0.05). Conclusion: Higher isolation rate of E. coli and pathogenic species of Staphylococci in raw milk samples collected from different milk cooperative centers in Hawassa could be associated to mastitis, poor udder preparation, poor milker’s hygiene, poor milk handling practices, poor environmental sanitation and sanitation of milking equipment. Overall, this study showed that pathogenic species of staphylococcus and E. coli are prevalent in raw milk produced and consumed in the area. Therefore, awareness should be created to producers and raw milk collecting cooperatives on strict preventive measures of raw milk contamination.

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1530
Author(s):  
Amanuel Balemi ◽  
Balako Gumi ◽  
Kebede Amenu ◽  
Sisay Girma ◽  
Muuz Gebru ◽  
...  

A study was carried out from August 2017 to February 2018 on lactating dairy cows, one-humped dromedary camels, and goats to determine mastitis in the Bule Hora and Dugda Dawa districts of in Southern Ethiopia. Milk samples from 564 udder quarters and udder halves from 171 animals consisting of 60 dairy cows, 51 camels, and 60 goats were tested for mastitis. Sixty-four positive udder milk samples were cultured, and bacterial mastitis pathogens were isolated and identified. The antibiotic resistance of bacterial isolates from milk with mastitis was tested against nine antimicrobials commonly used in the study area. Cow- and quarter-level prevalence of mastitis in dairy cows, camels, and goats was 33.3%, 26.3%, and 25% and 17.6%, 14.5%, and 20%, respectively. In cattle, the prevalence was significantly higher in Dugda Dawa than in Bule Hora. Major bacterial isolates were coagulase-negative Staphylococcus species (39.1%), S. aureus (17.2%), S. hyicus (14.1%), and S. intermedius and Escherichia coli (9.4% each). In camels, udder abnormality and mastitis were significantly higher in late lactation than in early lactation. Mastitis tends to increase with parity in camels. E. coli isolates were highly resistant to spectinomycin, vancomycin, and doxycycline, whereas most S. aureus isolates were multidrug-resistant. Most of the rural and periurban communities in this area consume raw milk, which indicates a high risk of infection with multidrug-resistant bacteria. We recommend a community-focused training program to improve community awareness of the need to boil milk and the risk of raw milk consumption.


2019 ◽  
Vol 40 (2) ◽  
pp. 723
Author(s):  
Amanda Bezerra Bertolini ◽  
Marcelo Augusto Orsi Dutra ◽  
Lívia Maísa Guiraldi ◽  
Wesley José dos Santos ◽  
Maria Izabel Merino de Medeiros ◽  
...  

Milk and its derivatives are good substrates for the proliferation of pathogenic and quality-deteriorating microorganisms, demanding rigorous care with milking, processing, and storage. Among the various bacteria that can grow in raw refrigerated milk, Yersinia enterocolitica, is an invasive enteropathogen of humans. This bacterium can cause a number of intestinal and extraintestinal clinical symptoms, ranging from mild gastroenteritis to mesenteric lymphadenitis, similar to appendicitis. To evaluate the prevalence of pathogenic Yersinia enterocolitica in raw milk from bulk milk tanks located in the State of São Paulo, 102 bovine milk samples (one per dairy farm) were evaluated by microbiological analyses, followed by biochemical tests PCR and genetic sequencing. Microbiological testing did not isolate Y. enterocolitica. However, PCR analysis revealed six samples that were positive for Y. enterocolitica (5.9%), confirmed by genetic sequencing. Only the inv gene was detected, which is present in virulent and avirulent Y. enterocolitica strains. There was great difficulty in microbiological isolation due to the difficulty of competitiveness of Y. enterocolitica in a very rich microbiota of raw milk. Although virulence genes known to be present in potentially pathogenic strains of Y. enterocolitica have not been identified, the presence of this pathogen in milk from expansion tanks, identified through PCR and confirmed by genetic sequencing, suggests that Y. enterocolitica may be a risk to public health, especially if milk and its derivatives are consumed without heat treatment.


2020 ◽  
pp. 126-133
Author(s):  
Omaima A. Madani ◽  
Telal M. Najeeb ◽  
Abdelhalim A. Hamza

This study was conducted to assess the factors influencing milk quality from dairy farmers in the Deam ALgarri aera in Sudan. A total of twenty row cow’s milk samples were collected from dairy farmers. The physicochemical parameters, bacteriological quality, milk adulteration and drug residues were assessed. The presence and enumeration of microorganisms was done based on Total Plate Count (TPC), Total Coliform Count (TCC), Total Staphylococcal count (TSC), and detection of pathogenic E. coli (O157: H7). The mean value of aerobic bacteria in the milk samples showed the higher TPC, TCC followed by TSC which were 111608 CFU/ml (log 4.95), 8145CFU/ml (log 3.42) and 6793.25 CFU/ml (log 3.29) respectively. The result of the detection of Escherichia coli O157:H7 which is a major foodborne pathogen causing severe disease in humans worldwide, all milk samples appear free from this pathogenic bacterium. The results of the physical properties of cow milk showed normal flavor, taste and colors, and higher pH (7.12), and total water (156%), the exception was the freezing point (- 0.444) and specific gravity while showed lower value (1.029) compared to SSMO limits but within FAO limits. The chemical properties of cow milk showed high amount of protein content (3.49%), lower solid not fat (SNF) which was (7.97%), Lactose (3.73%) and total solid (TS) (0.58%) when compared to both Sudanese Standard and Metrology Organization (SSMO) and Food and Agriculture Organization (FAO) standards. The result of presence or absence of antibiotic residue to the all milk samples was indicated absence antibiotic residues with (6 mm) and (7 mm) zone inhibitor against Bacillus subtilis. Isolation and identification of some pathogenic bacteria was performed according to standard methods, the results demonstrated the presence of food-borne microorganisms E. coli, klebsiela, sp, Pseudomonas sp, and Shiglla sp. with percentages of 31.1%, 29.7%, 38.4%, and 1% respectively, the presence of these bacteriological quality in the raw milk samples indicted an urgent need to implement good hygiene practices from farm to the consumer.


2021 ◽  
Vol 12 ◽  
Author(s):  
Yasmine H. Tartor ◽  
Norhan K. Abd El-Aziz ◽  
Rasha M. A. Gharieb ◽  
Hend M. El Damaty ◽  
Shymaa Enany ◽  
...  

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.


2018 ◽  
Vol 38 (8) ◽  
pp. 1577-1583 ◽  
Author(s):  
José C. Ribeiro Júnior ◽  
Pedro I. Teider Junior ◽  
André L.M. Oliveira ◽  
Edson A. Rios ◽  
Ronaldo Tamanini ◽  
...  

ABSTRACT: Pseudomonas, the main genus of gram-negative microorganisms isolated from milk, is psychrotrophic, biofilm-forming, and thermo-resistant deteriorating enzyme producers. The aim of this study was to quantify Pseudomonas spp. in goat’s and cow’s milk produced in the Paraná state, Brazil, to evaluate the deteriorating activity of the isolates at mesophilic and psychrotrophic conditions and to identify, at the species level, the isolates with alkaline metalloprotease (aprX gene) production potential. Microbiological, biochemical and molecular methods were used for isolating, confirming and identifying of isolates. The mean counts were 1.6 (±6.3)x104 and 0.89(±3)x102 CFU/mL for goat and bovine milk samples, respectively, immediately after milking. Of the Pseudomonas colonies isolated from goat milk (n=60), 91.7% showed proteolytic potential when incubated at 35°C/48 h and 80% at 7°C/10 days, and lipolytic potential was observed in 95% of the isolates incubated in mesophilic and 78.3% at refrigeration conditions. From the isolates of bovine milk (n=20), 35% showed proteolytic activity only when incubated at 35°C/48 h, and lipolytic potential was observed in 25% of the isolates incubated at 7°C/10d and 35°C/48h. It was observed that 83.3% and 25% of the isolates genetically confirmed as Pseudomonas spp. of goat and bovine milk showed the potential for alkaline metalloprotease production, with the species P. azotoformans, P. koreensis, P. gessardii, P. monteilii and P. lurida being the most frequent in goat milk and P. aeruginosa the only species identified in cow milk.


2008 ◽  
Vol 71 (5) ◽  
pp. 1023-1027 ◽  
Author(s):  
R. N. COBBOLD ◽  
M. A. DAVIS ◽  
D. H. RICE ◽  
M. SZYMANSKI ◽  
P. I. TARR ◽  
...  

A survey for Shiga toxigenic Escherichia coli in raw milk and beef was conducted within a defined geographic region of the United States. Prevalence rates based on detection of Shiga toxin gene (stx) were 36% for retail beef, 23% for beef carcasses, and 21% for raw milk samples, which were significantly higher than were Shiga toxigenic E. coli isolation rates of 7.5, 5.8, and 3.2%, respectively. Seasonal prevalence differences were significant for stx positivity among ground beef and milk samples. Distribution of stx subtypes among isolates varied according to sample type, with stx1 predominating in milk, stx2 on carcasses, and the combination of both stx1 and stx2 in beef. Ancillary virulence markers eae and ehx were evident in 23 and 15% of isolates, respectively. Pulsed-field gel electrophoresis demonstrated associations between food isolates and sympatric bovine fecal, and human clinical isolates. These data demonstrate that non-O157 Shiga toxigenic E. coli is present in the food chain in the Pacific Northwest, and its risk to health warrants critical assessment.


2017 ◽  
Vol 35 (No. 4) ◽  
pp. 311-320 ◽  
Author(s):  
Abusheliabi Aisha ◽  
Al-Holy Murad A ◽  
Al-Rumaithi Hind ◽  
Al-Khaldi Sufian ◽  
Al-Nabulsi Anas A ◽  
...  

The growth behaviour of foodborne pathogens (Staphylococcus aureus, Listeria monocytogenes, E. coli O157:H7 and Salmonella spp.) was investigated in pasteurised camel milk and compared with pasteurised bovine milk at different incubation temperatures. This study also aimed to compare the growth patterns of these four foodborne pathogens in pasteurised and raw camel milk. Pasteurised or raw camel milk and pasteurised bovine milk were separately inoculated with a cocktail of three strains of each foodborne pathogen. The inoculated milk samples were incubated at 10, 25, and 37°C. The total bacterial count (TBC) in raw milk and the total thermoduric bacteria count (TDB) in pasteurised milk samples were monitored. Greater growth inhibition rates of four pathogens were obtained for the pasteurised camel milk compared to the pasteurised bovine milk. Raw and pasteurised camel milk exerted bacteriostatic effect against all tested pathogens, particularly for the first 8 h of incubation in milk at the different temperatures. Pasteurised camel milk exerted an inhibitory activity that was equivalent to that of raw camel milk.


Author(s):  
E. Seker ◽  
H. Yardimci

Three hundred rectal faecal samples and 213 raw milk samples obtained from the tanks and containers were examined using standard cultural methods. Escherichia coli O157:H7 was isolated from 11 (3.7 %) of 300 faecal samples and 3 (1.4 %) of 213 raw milk samples. It was determined that 8 (73 %) of E. coli O157:H7 strains isolated from faecal samples originated from water buffaloes younger than 2 years of age and 3 (27 %) from 2-year-old and older water buffaloes. This is the 1st isolation of Escherichia coli O157:H7 from faecal and milk samples of water buffaloes in Turkey.


2021 ◽  
pp. 2410-2418
Author(s):  
Waleed Younis ◽  
Sabry Hassan ◽  
Hams M. A. Mohamed

Background and Aim: Raw milk is considered an essential source of nutrition during all stages of human life because it offers a valuable supply of protein and minerals. Importantly, milk is considered a good media for the growth and contamination of many pathogenic bacteria, especially food-borne pathogens such as Escherichia coli. Thus, the objective of this study was to characterize E. coli and detect its virulence factors and antibiotic resistance from raw milk samples. Materials and Methods: Raw milk samples (n=100) were collected from different localities in Qena, Egypt, and investigated for the presence of E. coli using different biochemical tests, IMViC tests, serotyping to detect somatic antigen type, and molecularly by polymerase chain reaction (PCR) tests. The presence of different virulence and antimicrobial genes (hly, eae, stx1, stx2, blaTEM, tetA(A), and tetB genes) in E. coli isolates was evaluated using PCR. Results: The results demonstrated that 10 out of 100 milk samples were contaminated with E. coli. Depending on serology, the isolates were classified as O114 (one isolate), O27 (two isolates), O111 (one isolate), O125 (two isolates), and untypeable (five isolates) E. coli. The sequencing of partially amplified 16S rRNA of the untypeable isolates resulted in one isolate, which was initially misidentified as untypeable E. coli but later proved as Enterobacter hormaechei. Moreover, antibacterial susceptibility analysis revealed that nearly all isolates were resistant to more than 3 families of antibiotics, particularly to β-lactams, clindamycin, and rifampin. PCR results demonstrated that all E. coli isolates showed an accurate amplicon for the blaTEM and tetA(A) genes, four isolates harbored eae gene, other four harbored tetB gene, and only one isolate exhibited a positive stx2 gene. Conclusion: Our study explored vital methods for identifying E. coli as a harmful pathogen of raw milk using 16S rRNA sequencing, phylogenetic analysis, and detection of virulence factors and antibiotic-resistant genes.


2019 ◽  
Vol 20 ◽  
Author(s):  
Laryssa F. Ribeiro ◽  
Mayhara M. C. Barbosa ◽  
Fernanda R. Pinto ◽  
Leticia F. Lavezzo ◽  
Gabriel A. M. Rossi ◽  
...  

Abstract This study focused on detecting diarrheagenic Escherichia coli, enteropathogenic E. coli (EPEC), Shiga-toxin-producing E. coli (STEC), enterohemorrhagic E. coli (EHEC or STEC:EPEC), enterotoxigenic E. coli (ETEC), and enteroaggregative E. coli (EAEC) in raw milk, water, and cattle feces sampled from non-technified dairy farms located in the northeastern São Paulo State, Brazil. Thirty-six water samples were collected at different points, namely, water wells (8 samples), water intended for human consumption (8 samples), water from milking parlor (8 samples), and water intended for animal consumption (7 samples), headwaters (1 sample), rivers (3 samples), and reservoirs (1 sample). Three raw milk samples were taken directly from bulk tanks in each farm, totalizing 24 samples. Feces samples were collected using rectal swabs from 160 bovines (20 animals per farm). E. coli was detected in 128 feces samples (80%), 16 raw milk samples (66.67%), and 20 water samples (55.56%). STEC (26 samples, 16.25%), EPEC (10 samples, 6.25%), STEC: EPEC (5 samples, 3.13%), and STEC: ETEC (1 sample, 0.63%) were the most prevalent strains detected in samples from cattle feces. EPEC, STEC, and STEC: EPEC strains were detected in 4.17% (1 sample), 16.67% (4 samples), and 4.17% (1 sample) of raw milk samples, respectively. STEC strains were detected in water used in the milking parlor, while no EAEC strain was detected. As a conclusion, cattle feces are important contamination sources of pathogenic E. coli in non-technified dairy farms and, consequently, cross-contamination among feces, water, and/or raw milk can occur. The use of quality water and hygienic practices during milking are recommended to avoid contamination since pathogens can be transmitted to humans via raw milk or raw milk cheese ingestion.


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