scholarly journals Pathogens isolated from bovine clinical mastitis and their antimicrobial resistance

2022 ◽  
Vol 78 (01) ◽  
pp. 6606-2022
Author(s):  
OZGE ARDICLI ◽  
SERPIL KAHYA DEMIRBILEK ◽  
TAYFUN CARLI K.
Keyword(s):  
Antimicrobial Resistance ◽  
Dairy Cattle ◽  
Pasteurella Multocida ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
Diffusion Method ◽  
Clinical Aspects ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Milk Samples

This study aimed to isolate aerobic and microaerophilic bacteria from mastitis milk samples, as well as to determine their antibiotic resistance. A total of 196 bovine mastitis milk samples were tested by standard bacteriological methods and with API identification test kits. Antimicrobial susceptibility testing was performed by the Kirby-Bauer disk diffusion method. The results revealed that the predominant isolate was S. aureus, with an isolation rate of 28%, followed by Streptococcus spp. (27%) and E. coli (19%). Isolation rates for Corynebacterium spp., Mycoplasma spp., and Pseudomonas aeruginosa were 11%, 6%, and 4%, respectively. Compared to the bacteria mentioned above, lower percentages were observed for Trueperella pyogenes (2%), Pasteurella multocida (2%), and Klebsiella pneumoniae (1%). A broad evaluation of antimicrobial resistance showed that the pathogens were resistant to tetracycline (68.63%), oxytetracycline (41.57%), ampicillin (39.08%), ceftiofur (38.1%), cephalexin (32.26%), penicillin (31.25%), amoxicillin/clavulanic acid (24.53%), enrofloxacin (24.44%), gentamycin (23.68%), and trimethoprim/sulfamethoxazole (22.09%). This study demonstrated that the sources of bacteria isolated from mastitis bovine milk samples were both contagious and environmental. More importantly, the present results demonstrate a critically high antimicrobial resistance in dairy cattle. For instance, E. coli isolates showed a crucial resistance to commonly used and recommended antimicrobials, including ceftiofur (100%), cephalexin (83.33%), and tetracycline (94.44%). The results of this study may provide valuable information about clinical aspects of bovine mastitis infections and current antimicrobial resistance levels in dairy cattle.

scholarly journals Molecular characterization of antimicrobial resistance and virulence genes of Escherichia coli isolates from bovine mastitis

2020 ◽  
Vol 13 (8) ◽  
pp. 1588-1593
Author(s):  
Zuhair Bani Ismail ◽  
Sameeh M. Abutarbush
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Shiga Toxin ◽  
Virulence Genes ◽  
Bovine Mastitis ◽  
Virulence Gene ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Procaine Penicillin

Background and Aim: Mastitis is a common and economically important disease in dairy cattle. It remains one of the most common reasons for the extensive use of antimicrobials in dairy farms leading to the emergence of antimicrobial-resistant pathogens. The aim of this study was to determine the patterns of antimicrobial resistance of Escherichia coli isolates from bovine mastitis and to identify prominent antimicrobial resistance and virulence genes among isolated strains. Materials and Methods: Antimicrobial susceptibility testing against six antibiotic groups, including tetracyclines, aminoglycosides, beta-lactams, macrolides, sulfonamides, and fluoroquinolones was performed using the disk diffusion method. PCR was performed on resistant isolates to detect resistance and virulence genes using commercially available primers. Results: Out of 216 milk samples cultured, 14 samples yielded E. coli isolates. All isolates (100%) were resistant to ampicillin, amoxicillin, procaine penicillin, streptomycin, oxytetracycline, and sulfamethoxazole-trimethoprim. Only one isolate (7%) was sensitive to gentamicin, and all isolates (100%) were sensitive to enrofloxacin and ciprofloxacin. All isolates carried at least one resistance gene against one or more of the major antibiotic groups. All isolates carried the ereA, tetG, tetE, and tetB genes, followed by tetA (93%), ampC (86%), strA (86%), sul1 (78%), tetD (71%), tetC (57%), aadA (57%), and strB (36%). The lowest percentage of isolates carried bla1 (17%) and bla2 (12%) genes, and none of the isolates carried the qnrA gene. Most of the isolates (93%) carried the Shiga toxin 1 virulence gene, followed by complement resistance protein (79%), intimin (64%), Shiga toxin 2 (36%), cytotoxic necrotizing factor (35%), aerotaxis receptor (21%), and type 1 fimbriae (15%). Conclusion: Results of this study indicate that the high percentages of E. coli isolate from bovine mastitis are resistant to two or more of the major antibiotic groups, irrespective of the presence or absence of relevant resistance or virulence genes.

scholarly journals Phenotypic Antimicrobial Susceptibility of Escherichia coli from Raw Meats, Ready-to-Eat Meats, and Their Related Samples in One Health Context

2021 ◽  
Vol 9 (2) ◽  
pp. 326
Author(s):  
Frederick Adzitey ◽  
Nurul Huda ◽  
Amir Husni Mohd Shariff
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Antimicrobial Susceptibility ◽  
Resistance Index ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Disk Diffusion Method ◽  
E Coli ◽  
The Usa ◽  
Raw Meats

Meat is an important food source that can provide a significant amount of protein for human development. The occurrence of bacteria that are resistant to antimicrobials in meat poses a public health risk. This study evaluated the occurrence and antimicrobial resistance of E. coli (Escherichia coli) isolated from raw meats, ready-to-eat (RTE) meats and their related samples in Ghana. E. coli was isolated using the USA-FDA Bacteriological Analytical Manual and phenotypic antimicrobial susceptibility test was performed by the disk diffusion method. Of the 200 examined meats and their related samples, 38% were positive for E. coli. Notably, E. coli was highest in raw beef (80%) and lowest in RTE pork (0%). The 45 E. coli isolates were resistant ≥ 50% to amoxicillin, trimethoprim and tetracycline. They were susceptible to azithromycin (87.1%), chloramphenicol (81.3%), imipenem (74.8%), gentamicin (72.0%) and ciprofloxacin (69.5%). A relatively high intermediate resistance of 33.0% was observed for ceftriaxone. E. coli from raw meats, RTE meats, hands of meat sellers and working tools showed some differences and similarities in their phenotypic antimicrobial resistance patterns. Half (51.1%) of the E. coli isolates exhibited multidrug resistance. The E. coli isolates showed twenty-two different resistant patterns, with a multiple antibiotic resistance index of 0.0 to 0.7. The resistant pattern amoxicillin (A, n = 6 isolates) and amoxicillin-trimethoprim (A-TM, n = 6 isolates) were the most common. This study documents that raw meats, RTE meats and their related samples in Ghana are potential sources of antimicrobial-resistant E. coli and pose a risk for the transfer of resistant bacteria to the food chain, environment and humans.

scholarly journals Molecular types, virulence profiles and antimicrobial resistance of Escherichia coli causing bovine mastitis

2019 ◽  
Vol 6 (1) ◽  
pp. e000369 ◽  
Author(s):  
Magdalena Nüesch-Inderbinen ◽  
Nadine Käppeli ◽  
Marina Morach ◽  
Corinne Eicher ◽  
Sabrina Corti ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Bovine Mastitis ◽  
Tetracycline Resistance ◽  
Diffusion Method ◽  
Sample Population ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Phylogenetic Grouping ◽  
Resistance Rates ◽  
Susceptibility Profiles

BackgroundEscherichia coli is an important aetiological agent of bovine mastitis worldwide.MethodsIn this study, 82 E. coli from bovine mastitis milk samples from 49 farms were analysed for their genetic diversity using phylogenetic grouping and multilocus sequence typing. The isolates were examined by PCR for a selection of virulence factors (VFs). Antimicrobial susceptibility profiles were assessed using the disk diffusion method.ResultsThe most prevalent phylogroups were group B1 (41.5 per cent of the isolates) and group A (30.5 per cent). A variety of 35 different sequence types (STs) were identified, including ST1125 (11 per cent), ST58 (9.8 per cent), ST10 (8.5 per cent) and ST88 (7.3 per cent). Aggregate VF scores (the number of unique VFs detected for each isolate) ranged from 1 to 3 for 63.4 per cent of the isolates and were at least 4 for 12.2 per cent. For 24.4 per cent of the isolates, the score was 0. The three most frequent VFs were traT, fyuA and iutA. The majority (72 per cent) of the isolates harboured traT. The majority (68.3 per cent) of the isolates were fully susceptible to all antimicrobials tested, with 22 per cent resistant to ampicillin and 14.6 per cent to tetracycline. Resistance rates were low for gentamicin (3.7 per cent), amoxicillin/clavulanic acid (2.4 per cent) and ceftiofur (1.2 per cent), respectively.ConclusionAmong the study’s sample population, E. coli strains were genotypically diverse, even in cows from the same farm, although some STs occurred more frequently than others. Susceptibility to clinically relevant compounds remained high.

scholarly journals Characterization of cefotaxime resistant Escherichia coli isolated from broiler farms in Ecuador

2018 ◽  
Author(s):  
Christian Vinueza-Burgos ◽  
David Ortega-Paredes ◽  
Cristian Narváez ◽  
Lieven De Zutter ◽  
Jeannete Zurita
Keyword(s):  
Antimicrobial Resistance ◽  
Latin American ◽  
Production Systems ◽  
Diffusion Method ◽  
Poultry Production ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Resistance Patterns ◽  
Latin American Countries

AbstractAntimicrobial resistance (AR) is a worldwide concern. Up to a 160% increase in antibiotic usage in food animals is expected in Latin American countries. The poultry industry is an increasingly important segment of food production and contributor to AR. The objective of this study was to evaluate the prevalence, AR patterns and the characterization of relevant resistance genes in Extended Spectrum β-lactamases (ESBL) and AmpC E. coli from large poultry farms in Ecuador. Sampling was performed from June 2013 to July 2014 in 6 slaughterhouses that slaughter broilers from 115 farms totaling 384 flocks. Each sample of collected caeca was streaked onto TBX agar supplemented with cefotaxime (3 mg/l). In total, 176 isolates were analyzed for antimicrobial resistance patterns by the disk diffusion method and for blaCTX-M, blaTEM, blaCMY, blaSHV, blaKPC, and mcr-1 by PCR and sequencing. ESBL and AmpC E. coli were found in 362 flocks (94.3%) from 112 farms (97.4%). We found that 98.3% of the isolates were multi-resistant to antibiotics. Low resistance was observed for ertapenem and nitrofurantoin. The most prevalent ESBL genes were the blaCTX-M (90.9%) blaCTX-M-65, blaCTX-M-55 and blaCTX-M-3 alleles. Most of the AmpC strains presented the blaCMY-2 gene. Three isolates showed the mcr-1 gene. Poultry production systems represent a hotspot for antimicrobial resistance in Ecuador, possibly mediated by the extensive use of antibiotics. Monitoring this sector in national and regional plans of antimicrobial resistance surveillance should therefore be considered.

scholarly journals 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

2021 ◽  
Vol 12 ◽  
Author(s):  
Yasmine H. Tartor ◽  
Norhan K. Abd El-Aziz ◽  
Rasha M. A. Gharieb ◽  
Hend M. El Damaty ◽  
Shymaa Enany ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Klebsiella Pneumoniae ◽  
Genome Sequencing ◽  
Bovine Milk ◽  
Bovine Mastitis ◽  
Raw Milk ◽  
Whole Genome ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
E Coli

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.

scholarly journals Screening commercial teat disinfectants against bacteria isolated from bovine milk using disk diffusion

2019 ◽  
Vol 12 (5) ◽  
pp. 629-637 ◽  
Author(s):  
Sarah Rose Fitzpatrick ◽  
Mary Garvey ◽  
Kieran Jordan ◽  
Jim Flynn ◽  
Bernadette O'Brien ◽  
...  
Keyword(s):  
Bacterial Species ◽  
Screening Method ◽  
Bovine Milk ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Bacterial Strains ◽  
Disk Diffusion Method ◽  
Associated Bacteria ◽  
Bacterial Inhibition ◽  
Milk Samples

Background and Aim: Teat disinfection is an important tool in reducing the incidence of bovine mastitis. Identifying the potential mastitis-causing bacterial species in milk can be the first step in choosing the correct teat disinfectant product. The objective of this study was to screen commercial teat disinfectants for inhibition against mastitis-associated bacteria isolated from various types of milk samples. Materials and Methods: Twelve commercially available teat disinfectant products were tested, against 12 mastitis-associated bacteria strains isolated from bulk tank milk samples and bacterial strains isolated from clinical (n=2) and subclinical (n=3) quarter foremilk samples using the disk diffusion method. Results: There was a significant variation (7-30 mm) in bacterial inhibition between teat disinfection products, with products containing a lactic acid combination (with chlorhexidine or salicylic acid) resulting in the greatest levels of bacterial inhibition against all tested bacteria (p<0.05). Conclusion: In this study, combined ingredients in teat disinfection products had greater levels of bacterial inhibition than when the ingredients were used individually. The disk diffusion assay is a suitable screening method to effectively differentiate the bacterial inhibition of different teat disinfectant products.

scholarly journals In vitro Synergistic Antimicrobial Activity of Romanian Propolis and Antibiotics against Escherichia coli Isolated from Bovine Mastitis

2015 ◽  
Vol 43 (2) ◽  
pp. 327-334 ◽  
Author(s):  
Mihaela NICULAE ◽  
Laura STAN ◽  
Emoke PALL ◽  
Anamaria Ioana PAȘTIU ◽  
Iulia Maria BALACI ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Chemical Composition ◽  
Bovine Mastitis ◽  
Antibacterial Properties ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Ethanolic Extracts

The study was aimed to characterize the chemical composition and the antimicrobial activity of Romanian propolis ethanolic extracts (EEP) against antibiotic-sensitive and antibiotic-resistant E. coli strains isolated from bovine mastitis. The preliminary antimicrobial screening was performed by a disk diffusion method, followed by determination of minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) based on broth microdilution assay; further, the synergistic action of propolis with antimicrobial drugs was assessed by a disk diffusion method on agar containing subinhibitory concentrations of propolis. For the chemical characterisation of EEP, the flavonoids (flavones/flavonols, flavanones/dihydroflavonols) and total phenolics were evaluated by spectrophotometric methods. The phenolic compounds of these extracts were also determined using HPLC. The results indicated for Romanian propolis ethanolic extracts the typical poplar composition profile with flavonoids and phenolic acids as main biological active compounds, with chromatographic analysis data confirmed also spectrophotometrically. In addition, positively correlated with the chemical composition, a strong antimicrobial efficacy was exhibited towards E. coli strains, along with interesting synergistic interaction with antibiotics that can be further investigated to obtain propolis-based formulation with antibacterial properties. Subsequent in vitro and in vivo studies evaluating the safety and efficacy are intended to consider propolis in veterinary therapeutic protocols.

scholarly journals Detection of TEM and CTX-M Genes in Escherichia coli Isolated from Clinical Specimens at Tertiary Care Heart Hospital, Kathmandu, Nepal

Diseases ◽  
2021 ◽  
Vol 9 (1) ◽  
pp. 15
Author(s):  
Ram Shankar Prasad Sah ◽  
Binod Dhungel ◽  
Binod Kumar Yadav ◽  
Nabaraj Adhikari ◽  
Upendra Thapa Shrestha ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Clinical Isolates ◽  
Diffusion Method ◽  
Gram Negative Bacteria ◽  
Bacterial Isolates ◽  
Disk Diffusion Method ◽  
Gram Negative ◽  
Clinical Specimens ◽  
E Coli ◽  
Esbl Producers

Background: Antimicrobial resistance (AMR) among Gram-negative pathogens, predominantly ESBL-producing clinical isolates, are increasing worldwide. The main aim of this study was to determine the prevalence of ESBL-producing clinical isolates, their antibiogram, and the frequency of ESBL genes (blaTEM and blaCTX-M) in the clinical samples from patients. Methods: A total of 1065 clinical specimens from patients suspected of heart infections were collected between February and August 2019. Bacterial isolates were identified on colony morphology and biochemical properties. Thus, obtained clinical isolates were screened for antimicrobial susceptibility testing (AST) using modified Kirby–Bauer disk diffusion method, while ESBL producers were identified by using a combination disk diffusion method. ESBL positive isolates were further assessed using conventional polymerase chain reaction (PCR) to detect the ESBL genes blaTEM and blaCTX-M. Results: Out of 1065 clinical specimens, 17.8% (190/1065) showed bacterial growth. Among 190 bacterial isolates, 57.4% (109/190) were Gram-negative bacteria. Among 109 Gram-negative bacteria, 40.3% (44/109) were E. coli, and 30.2% (33/109) were K. pneumoniae. In AST, 57.7% (n = 63) Gram-negative bacterial isolates were resistant to ampicillin and 47.7% (n = 52) were resistant to nalidixic acid. Over half of the isolates (51.3%; 56/109) were multidrug resistant (MDR). Of 44 E. coli, 27.3% (12/44) were ESBL producers. Among ESBL producer E. coli isolates, 58.4% (7/12) tested positive for the blaCTX-M gene and 41.6% (5/12) tested positive for the blaTEM gene. Conclusion: Half of the Gram-negative bacteria in our study were MDR. Routine identification of an infectious agent followed by AST is critical to optimize the treatment and prevent antimicrobial resistance.

scholarly journals Surveillance of antimicrobial resistance of maltose negative Staphylococcus aureus in South African dairy herds

2020 ◽  
Author(s):  
Joanne Karzis ◽  
Inge-Marié Petzer ◽  
Edward F. Donkin ◽  
Vinny Naidoo ◽  
Eric M. C. Etter
Keyword(s):  
South Africa ◽  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Dairy Herds ◽  
Disk Diffusion Method ◽  
Milk Samples ◽  
Kwazulu Natal ◽  
Over Time

Abstract Background The discovery of antimicrobials in the 1930s was one of the greatest achievements in medicine. However, bacterial resistance to antimicrobials was already observed in the 1940s and has been reported since then in both human and veterinary medicine, including in dairy cows. Many years of monitoring milk samples in South Africa, has led to the identification of a new strain of Staphylococcus aureus (S. aureus), which is maltose negative and appears to be an emerging pathogen. In this study the differences in susceptibility to antimicrobials of this strain were evaluated over time, over different seasons, in different provinces, and according to somatic cell count (SCC) categories. Results A data set of 271 maltose negative S. aureus isolates, cultured from milk samples from 117 herds out of the estimated 2000 commercial dairy herds in South Africa between 2010 and 2017, was studied using the disk diffusion method. This analysis was done using the Clinical Laboratory Standards Institute (CLSI) breakpoints in order to compare using both the previous (Intermediate category grouped with Resistant) and current definitions, (Intermediate category grouped with Susceptible). The results of the analysis between the previous and the current definitions differed for tylosin, cefalonium, oxy-tetracycline and cloxacillin. Neither the analysis using the previous nor the current systems showed an effect of province for the maltose negative S. aureus. This was in contrast to the results for maltose positive S. aureus where differences between provinces were shown in a previous study, with the lowest prevalence of resistance shown in KwaZulu-Natal during spring. For the susceptibility testing of 57 maltose negative and 57 maltose positive S. aureus isolates from 38 farms, from KwaZulu Natal, Eastern Cape and Western Cape. The minimum inhibitory concentration (MIC) results for the maltose negative S. aureus isolates confirmed the results of the disk diffusion method. Conclusions The maltose negative strains of S. aureus differed in general, in their antimicrobial resistance patterns over time, in comparison to maltose-positive S. aureus strains. MIC testing also indicated that more multidrug -resistant isolates were seen with the maltose negative S. aureus than in the maltose positive strains.

scholarly journals Isolation of Escherichia coli, Klebsiella spp. and Staphylococcus spp. from Bovine Mastitic Milk in Nuwera Eliya District of Sri Lanka and their Sensitivity to Commonly Used Antibiotics

2018 ◽  
pp. 1-13
Author(s):  
T. S. P. J. Jayaweera ◽  
J. L. P. C. Randika ◽  
H. G. C. L. Gamage ◽  
N. N. Udawatta ◽  
W. U. N. T. S. Ellepola ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Sri Lanka ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Milk Samples ◽  
Staphylococcus Spp ◽  
Klebsiella Spp ◽  
Clinical Cases

Aims: Mastitis is one of the very important and most common diseases among dairy cattle globally which leads to severe economical losses in the dairy industry. For the sustainability of the dairy sector it is critical that efficient, economically feasible treatment regime is available for clinical cases of mastitis as a part of the control program with minimum risk for residues in milk. Antimicrobials are the most common drugs of choice for controlling and preventing this devastating condition. But the frequent use of antibiotics leads to the development of resistant bacteria which could have an adverse effect on human health as well. To mitigate this destructive constraint in the industry, identifying the etiology and their susceptibilities to remedial measures are of paramount importance. Hence this study was aimed at isolating and identifying the common bacterial etiology Escherichia coli, Klebsiella spp. and Staphylococcus spp. of mastitis and evaluating the antimicrobial susceptibility of the isolates in order to develop mastitis control strategies in the area. Study Design: Milk samples were collected from mastitic cows in different stages including subclinical and clinical cases based on the results of California Mastitis Test Place and Duration of Study: Samples were collected from dairy farms in Nuwera Eliya District, Sri Lanka and Laboratory investigations were carried out in the Laboratory of Livestock Production, Faculty of Agricultural Sciences, Sabaragamuwa University of Sri Lanka, Between Aug. 2017 and Nov. 2017. Methodology: E.coli, Klebsiella spp. and Staphylococcus spp. were isolated from 31 milk samples and susceptibility to commonly used antibiotics (Trimethoprim, Oxytetracycline, Chloramphenicol, Cephalexin, Enrofloxacin and Ciprofloxacin) was determined by Kirby Bauer disk diffusion method. Results: The study revealed that the most common isolate was the Klebsiella spp. and it is 54.8% and other two organisms Staphylococcus spp. had 51.6% and Escherichia coli 41.9%. Of all isolated pathogen, 97.1% exhibited resistant to Cephalexin and it was the highest while lowest resistance was to Chloramphenicol (31.4%). Among the other antibiotics, 54.3% of total isolates showed resistance to Trimethoprim followed by 42.9% to Oxytetracycline and Enrofloxacin, 34.3% was resistant to Ciprofloxacin. Resistance to at least one antibiotic was observed for the isolated microorganisms. All the three isolated pathogens are more resistant to Cephalexin. Both E. coli and Klebsiella spp. show 100% resistance to Cephalexin while Staphylococci had 92.9% resistance. This further revealed that E. coli (10%) and Klebsiella spp. (27.3%) are showing the least resistance to Chloramphenicol, None of Staphylococcus spp. (0%) isolated show resistance to Enrofloxacin. Conclusion: Most common organisms isolated were Klebsiella spp. followed by Staphylococcus spp., E. coli and there is a resistance of isolated organisms to some commonly used antibiotics.

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