Isolation and Identification of Staphylococcus Aureus from Dairy Farms in Bishoftu Town, Ethiopia

Mapping Intimacies â—½  
2021 â—½  
Author(s):  
Sara Amanuel Bude â—½  
Abdi Kidane Mengesha
Keyword(s):  
Staphylococcus Aureus â—½  
Dairy Farms â—½  
Nutrient Agar â—½  
Blood Agar â—½  
Swab Sample â—½  
Biochemical Tests â—½  
Isolation And Identification â—½  
Mannitol Salt Agar â—½  
Milk Samples â—½  
Golden Yellow

Abstract A cross-sectional study was conducted from February, 2020 to March, 2020 in selected dairy farms in Bishoftu Town to isolate and identify Staphylococcus aureus from milk samples, swab (teat swab, tank swab and bucket swab) samples. The samples were transported to microbiology laboratory and, isolation and identification of an organism was based on morphological, cultural and biochemical characteristics. Accordingly a total of 120 samples of them 58 milk sample, 58 polled teat swab sample, 2 polled tank swab sample and 2 polled bucket swab samples. Those milk samples and swab sample were collected and cultured parallel on both Blood agar and nutrient agar. Out of total samples 68/120 were well grown on both Medias and sub cultured on nutrient agar for farther identification using Primary and secondary biochemical tests. Accordingly, 68 milk samples were shown typical large, round, golden yellow colonies with hemolysis when grown on blood agar, round shaped grape like clusters are seen under microscope after stained with grams stain, and catalase positive Staphylococcus aureus with bubble formation is observed and typical yellow pigmentation on mannitol salt agar which is selective media for genus Staphylococcus. Finally, 68(56.67%) Staphylococcus aureus were identified with coagulase test with the clumping result observed. Hence, implementing hygiene conditions, creation of awareness on control and prevention of subclinical mastitis in dairy farms and conducting drug sensitivity test for Staphylococcus aureus is recommended.

scholarly journals CHARACTERIZATION OF BACTERIA ASSOCIATED WITH PNEUMONIA IN BLACK BENGAL GOATS

2012 â—½  
Vol 9 (1) â—½  
pp. 67-71 â—½  
Author(s):  
MA Momin â—½  
MA Islam â—½  
MM Khatun â—½  
MM Rahman â—½  
MA Islam
Keyword(s):  
Antimicrobial Agents â—½  
Research Work â—½  
Nutrient Agar â—½  
Blood Agar â—½  
Biochemical Tests â—½  
Highly Sensitive â—½  
Golden Yellow â—½  
Agar Media â—½  
Staphylococcus Spp

The present research work was undertaken for the characterization of the bacterial pathogens responsible for pneumonia in black Bengal goats. Nasal swab samples (n = 50) were collected from the pneumonic black Bengal goats in Mymensingh and Sirajgonj districts. Samples were inoculated onto nutrient agar, eosin methylene blue (EMB) agar, MacConkey agar, and blood agar media for isolation of bacteria. Identification of bacteria was performed by the Gram's staining method, cultural properties and biochemical tests. Antibiotic sensitivity of bacterial isolates was performed against 11 antimicrobial agents. Pasteurella spp were isolated from 25 cases, and Staphylococcus spp from 13 cases. Mixed infection caused by the Pasteurella spp and Staphylococcus spp. were recorded in 12 cases. Pasteurella spp produced whitish, opaque circular and translucent colonies on nutrient agar, smooth, convex, glistening colonies on EMB agar and no hemolysis on blood agar. Staphylococcus spp have shown gray white or golden yellowish colonies on  nutrient agar. Golden yellow colonies without hemolysis or whitish colonies with hemolysis were also produced by Staphylococcus spp. on the blood agar media. Pasteurella spp were indole positive, MR-VP negative and ferment dextrose, sucrose and mannitol with the production of acid. The Staphylococcus spp were positive to MR-VP, coagulase and catalase reactions, negative to indole test and fermented five basic sugars with acid production. Results of cultural and biochemical tests supported that these two isolates belonged to P. multocida and S. aureus. P. multocida were highly sensitive to ciprofloxacin and resistant to penicillin. S. aureus found to be highly sensitive to erythromycin, tetracycline, enrofloxacin, and norfloxacin and less sensitive to amoxicillin. DOI = http://dx.doi.org/10.3329/bjvm.v9i1.11215Bangl. J. Vet. Med. (2011). 9(1): 67-71 

scholarly journals Antimicrobial Resistance Profiles of Staphylococcus Aureus Isolated from Meat Carcasses and Bovine Milk in Abattoirs and Dairy Farms of the Eastern Cape, South Africa

2018 â—½  
Author(s):  
Ezekiel Green â—½  
Abongile Pekana
Keyword(s):  
South Africa â—½  
Staphylococcus Aureus â—½  
Bovine Milk â—½  
Dairy Farms â—½  
Penicillin G â—½  
Eastern Cape Province â—½  
Eastern Cape â—½  
Biochemical Tests â—½  
Milk Samples â—½  
Antibiotic Susceptibility Test

Background: Staphylococcus aureus (S. aureus) occasionally threatens the life of the host as a persistent pathogen even though it is normal flora of humans and animals. We characterized drug resistance in S. aureus isolated from animal carcasses and milk samples from the abattoirs and dairy farms in the Eastern Cape Province. Methods: A 1000 meat swab samples and 200 raw milk samples were collected from selected abattoirs and dairy farms in the Eastern Cape Province, South Africa. S. aureus was isolated and positively identified using biochemical tests and confirmed by molecular methods. Antibiotic susceptibility test against 14 different antibiotics was performed against all isolates. Antibiotic resistance genes were also detected. Results: Of the 1200 samples collected, 134 (11.2%) samples were positive for S. aureus. Resistance ranged from 71.6% for penicillin G to 39.2% for tetracycline. Resistance gene (blaZ) was detected in 13 (14.9%), while msrA was found in 31 (52.5%) of S. aureus isolates. Conclusions: The present result shows the potential dissemination of multidrug-resistant S. aureus strains in the dairy farms and abattoirs in the Eastern Cape. Therefore, this implies that the organism may rapidly spread through food and pose serious public health risk

scholarly journals Antibacterial Activity Of Bee Venom Against Multidrug Resistance Staphylococcus Aureus From Milk Of Cow And Buffaloes

2020 â—½  
Vol 11 (2) â—½  
pp. 2177-2182
Author(s):  
Adnan Kamel Shebeeb â—½  
Amaal Fadhil Ghanim â—½  
Hussam Sami Awayid
Keyword(s):  
Staphylococcus Aureus â—½  
Antibacterial Activity â—½  
Cow Milk â—½  
Diffusion Method â—½  
Efficiency Ratio â—½  
Technical Institute â—½  
Biochemical Tests â—½  
Isolation And Identification â—½  
Bacteriological Study â—½  
Milk Samples

Bacteriological study includes bacteria isolated from Buffaloes milk, and Cow at Wassit Province was done in Technical institute. 120 milk samples were collected randomly from different places in Wassit Province included 60 samples from Buffaloes milk and 60 samples from Cow milk during the period October to December 2018.A bacteriological study was conducted for isolation and identification S. aureus by morphological and biochemical tests. The results showed that (20%) of Buffaloes milk and (13.33%) in Cow milk (13.33 %) of Cow milk contaminated with S. aureus. Antibiogram pattern of  S. aureus  was carried out by using  a diffusion method from an antibiotic saturated disk , Results were appeared the more effect in S. aureus  isolated from Buffaloes milk  was COT in efficiency ratio ( 78.33 % ), CLR in efficiency ratio   ( 61 % ), SPX in efficiency ratio ( 51.66 % ), and L in efficiency ratio ( 15 % ) , Also the results appeared more effect in S. aureus isolated from Cow milk  was  COT in efficiency ratio ( 56.66 % ), CLR in efficiency ratio ( 35 % ), SPX in efficiency ratio      ( 30 % ), and L in efficiency ratio ( 8.33 % ), while S. aureus    appeared resistant in all milk samples from Buffaloes and Cow for antibiotic OX (100%) ,also MIC to staphylococcal ( 0.70 - 3.10 μg/ml ) While MBC concentration was(0.12-0.101 μg/ml).  

scholarly journals Antimicrobial Resistance Profiles of Staphylococcus aureus Isolated from Meat Carcasses and Bovine Milk in Abattoirs and Dairy Farms of the Eastern Cape, South Africa

2018 â—½  
Vol 15 (10) â—½  
pp. 2223 â—½  
Author(s):  
Abongile Pekana â—½  
Ezekiel Green
Keyword(s):  
Staphylococcus Aureus â—½  
Bovine Milk â—½  
Dairy Farms â—½  
Raw Milk â—½  
Multidrug Resistant â—½  
Penicillin G â—½  
Eastern Cape â—½  
Biochemical Tests â—½  
Milk Samples â—½  
Antibiotic Susceptibility Test

Background: Staphylococcus aureus (S. aureus) occasionally threatens the life of the host as a persistent pathogen even though it is normal flora of humans and animals. We characterized drug resistance in S. aureus isolated from animal carcasses and milk samples from the abattoirs and dairy farms in the Eastern Cape Province. Methods: A total of 1000 meat swab samples and 200 raw milk samples were collected from selected abattoirs and dairy farms. S. aureus was isolated and positively identified using biochemical tests and confirmed by molecular methods. An antibiotic susceptibility test was performed on all isolates for 14 antibiotics and correspondent genes were detected. Results: Of the 1200 samples collected, 134 (11.2%) samples were positive for S. aureus. Resistance ranged from 71.6% for penicillin G to 39.2% for tetracycline. A resistance gene (blaZ) was detected in 13 (14.9%), while msrA was found in 31 (52.5%) of S. aureus isolates. Conclusions: The present result shows the potential dissemination of multidrug-resistant S. aureus strains in the dairy farms and abattoirs in the Eastern Cape. Therefore, this implies that the organism may rapidly spread through food and pose serious public health risk.

scholarly journals Isolation and Identification of Staphyococci from Raw Milk of Cows Infected with Mastitis

2003 â—½  
Vol 27 (1) â—½  
pp. 174-179
Author(s):  
Safana, A.S. Al-Taan
Keyword(s):  
Staphylococcus Aureus â—½  
Raw Milk â—½  
Selective Medium â—½  
Isolation And Identification â—½  
Mannitol Salt Agar â—½  
Staphylococcal Species â—½  
Staph Aureus â—½  
Milk Samples â—½  
Positive Results â—½  
Prevalent Pathogen

Twenty-six isolates of staphylococci were recovered from of 100 raw milk samples collected from one hundred cows infected with mastitis. For the isolation of bacteria, Mannitol salt agar was used as a selective medium. Staphylococcal species were identified by API-STAPH system. The identified species were: - Staphy aureus (13), Staph. epidermidis(9), Staph. haemolyticus(2), Staph. lugdunensis()), and Staph. hominis). Staphylococcus aureus was the most prevalent pathogen among other Staphyloccocal species isolated in this study. Staph. aureus isolates revealed positive results for the defection of capsule structure and for production of some essential enzymes such as coagulase, phosphatase, DNA ase and haemolysin which are highly associated with the virulance of bacteria. Staph. aureus isolates were tested for their sensitivity to the antibiotics and the results were: 13(100%) isolates were sensitive to Gentamycin and Tetracyclene, 11 (84.6%) isolates were sensitive to Oxacillin, Penicillin, Ampicillin, Erythromycin and Cephalexin, where as only 2 (15.4%) isolates were Methicillin Resistant Staph. aureus (MRSA) which showed multi -resistant  towards many antibiotics used in this work.

scholarly journals Prevalence of methicillin-resistant Staphylococcus aureus and pattern of antimicrobial resistance in mastitis milk of cattle in Chitwan, Nepal

2021 â—½  
Vol 17 (1) â—½  
Author(s):  
Asmita Shrestha â—½  
Rebanta Kumar Bhattarai â—½  
Himal Luitel â—½  
Surendra Karki â—½  
Hom Bahadur Basnet
Keyword(s):  
Public Health â—½  
Staphylococcus Aureus â—½  
Antimicrobial Resistance â—½  
Subclinical Mastitis â—½  
World Health â—½  
Dairy Herds â—½  
Microbiological Analysis â—½  
Biochemical Tests â—½  
Methicillin Resistant â—½  
Milk Samples

Abstract Background The threat of methicillin-resistant Staphylococcus aureus (MRSA) exists globally and has been listed as a priority pathogen by the World Health Organization. One of the sources of MRSA emergence is livestock and its products, often raised in poor husbandry conditions. There are limited studies in Nepal to understand the prevalence of MRSA in dairy animals and its antimicrobial resistance (AMR) profile. A cross-sectional study was conducted in Chitwan, one of the major milk-producing districts of Nepal, from February 2018 to September 2019 to estimate the prevalence of MRSA in milk samples and its AMR profile. The collected milk samples (n = 460) were screened using the California Mastitis Test (CMT) and positive samples were subjected to microbiological analysis to isolate and identify S. aureus. Polymerase Chain Reaction (PCR) was used to identify the presence of the mecA gene and screen for MRSA. Results In total, 41.5% (191/460) of milk samples were positive in the CMT test. Out of 191 CMT positive milk samples, the biochemical tests showed that the prevalence of S. aureus was 15.2% (29/191). Among the 29 S. aureus isolates, 6.9% (2/29) were identified as MRSA based on the detection of a mecA gene. This indicates that that 1.05% (2/191) of mastitis milk samples had MRSA. The antibiotic sensitivity test showed that 75.9% (22/29) and 48.3% (14/29) S. aureus isolates were found to be sensitive to Cefazolin and Tetracycline respectively (48.3%), whereas 100% of the isolates were resistant to Ampicillin. In total 96.6% (28/29) of S. aureus isolates were multidrug-resistant (MDR). Conclusions This study revealed a high prevalence of S. aureus-mediated subclinical mastitis in dairy herds in Chitwan, Nepal, with a small proportion of it being MRSA carrying a mecA gene. This S. aureus, CoNS, and MRSA contaminated milk poses a public health risk due to the presence of a phenotype that is resistant to very commonly used antibiotics. It is suggested that dairy herds be screened for subclinical mastitis and treatments for the animals be based on antibiotic susceptibility tests to reduce the prevalence of AMR. Furthermore, future studies should focus on the Staphylococcus spp. to explore the antibiotic resistance genes in addition to the mecA gene to ensure public health.

scholarly journals Case Study: Misdiagnosis of Nonhemolytic Staphylococcus aureus Isolates from Cases of Bovine Mastitis as Coagulase-Negative Staphylococci

Animals â—½  
10.3390/ani11020252 â—½  
2021 â—½  
Vol 11 (2) â—½  
pp. 252
Author(s):  
Valerie E. Ryman â—½  
Felicia M. Kautz â—½  
Steve C. Nickerson
Keyword(s):  
Staphylococcus Aureus â—½  
Bovine Mastitis â—½  
Dairy Farm â—½  
Blood Agar â—½  
Coagulase Negative Staphylococci â—½  
Mannitol Salt Agar â—½  
Lactating Cows â—½  
Presumptive Identification â—½  
Microbiological Techniques

Staphylococcus aureus is one of the most concerning mastitis-causing pathogens in dairy cattle. Using basic microbiological techniques, S. aureus is typically identified by colony characteristics and hemolysis on blood agar where isolates without hemolysis are typically considered to be coagulase-negative staphylococci (CNS) isolates. Herein, we present a decade-long case study where suspected S. aureus isolates from one Georgia dairy farm were further tested to confirm presumptive identification. Presumptive identification of bacterial growth from 222 mammary secretions from bred Holstein heifers and lactating cows was conducted at the time of collection. Presumptive identification of S. aureus on blood agar was based on observation of colony morphology, color, and presence or absence of a broad zone of incomplete hemolysis and a smaller zone of complete hemolysis at 48 h. Those without hemolysis were presumptively characterized as CNS. All isolates were further plated on mannitol salt agar and a coagulase test was performed. A positive for both of these tests together was deemed to be S. aureus. A selection of isolates was tested using API® Staph to biochemically confirm S. aureus identification. Data showed that 63.96% of isolates presumed to be CNS isolates were identified as S. aureus, 9.46% of isolates presumed to be CNS isolates were identified as coagulase-positive staphylococci (CPS) species (but not S. aureus), and 26.58% of samples that were presumed to be CNS isolates were identified correctly.

scholarly journals Molecular characterization of virulence factors in Staphylococcus aureus isolated from bovine subclinical mastitis in central Ethiopia

2021 â—½  
Vol 71 (1) â—½  
Author(s):  
Desiye Tesfaye Tegegne â—½  
Gezahegne Mamo â—½  
Hika Waktole â—½  
Yohannes Equar Messele
Keyword(s):  
Staphylococcus Aureus â—½  
Dairy Cows â—½  
Virulence Factors â—½  
Virulence Genes â—½  
Animal Health â—½  
Subclinical Mastitis â—½  
Biochemical Tests â—½  
Milk Samples â—½  
Lactating Dairy Cows â—½  
Genomic Study

Abstract Purpose Staphylococcus aureus (S. aureus) is the most important pathogen involved in bovine mastitis in dairy production. S. aureus produces a spectrum of extracellular protein toxins and virulence factors which are thought to contribute to the pathogenicity of the organism. The aim of this work was to isolate and molecular characterize S. aureus associated with bovine subclinical mastitis in the central part of Ethiopia. Methods A total of 265 lactating dairy cows from various dairy farms in four different geographical locations were screened by the California mastitis test (CMT) for bovine subclinical mastitis. One hundred thirty CMT-positive milk samples were collected and transported to the laboratory. Different biochemical tests and polymerase chain reaction (PCR) were used for the identification of S. aureus isolates. Finally, PCR was performed for molecular detection of virulence genes. Results From a total of 265 lactating dairy cows screened, 49% (n = 130) were positive for bovine subclinical mastitis. One hundred thirty mastitic milk samples were subjected to bacterial culturing, and one hundred (76%) S. aureus isolates were identified based on phenotypic characters. Sixty-eight confirmed S. aureus isolates were obtained using PCR. The confirmed S. aureus isolates were tested for six virulence genes (tsst-1, hlb, eta, sea, clfA, and icaD) using PCR. Of the six virulence genes screened from all the isolates, only two (clfA and eta) were detected in the isolates. Out of 68 isolates, 25% and 22% were possessed the eta and clfA genes, respectively. Conclusion The presence of Staphylococcus aureus having virulence genes (eta and clfA) revealed that mastitis is a major concern nowadays affecting animal health, milk quality, and yield. Further genomic study of these isolates will provide broad new insights on virulence.

scholarly journals Identification and antibiogram study of bacterial species isolated from milk samples of different locations in Bangladesh

2016 â—½  
Vol 1 (3) â—½  
pp. 457-462 â—½  
Author(s):  
Md Nuruzzaman Munsi â—½  
Nathu Ram Sarker â—½  
Razia Khatun â—½  
Mohammed Khorshed Alam
Keyword(s):  
Escherichia Coli â—½  
Staphylococcus Aureus â—½  
Pathogenic Bacteria â—½  
Bacterial Species â—½  
Salmonella Typhi â—½  
Diffusion Method â—½  
Biochemical Tests â—½  
Public Health Importance â—½  
Milk Samples â—½  
First Choice

CowÂ’s milk containing pathogenic bacteria is an important threat to the consumers. The objectives of the present study were to identify the bacterial agents of public health importance in milk samples (n=35) of different locations and to determine their sensitivity to different antibiotics. The milk samples were collected and transported aseptically and subsequently allowed for culture in bacteriological media, GramÂ’s staining and biochemical tests for the identification of bacterial species. The bacteria identified were Staphylococcus aureus, Escherichia coli and Salmonella typhi, and their prevalence, in case of vendor milk specimens (n=28), were 96.43%, 53.57% and 35.71% respectively, and of brand milk specimens (n=7), were 42.86 %, 28.57% and 0%, respectively. This suggests that cautionary measures should be taken for quality milk production and consumption. The antibiotic sensitivity test was done by disc diffusion method and the average inhibition zones, in case of Staphylococcus aureus, were 32 mm for oxytetracycline, 26 mm for amoxicillin, 35 mm for ciprofloxacin, 27 mm for cefotaxime, 30 mm for ceftriaxone, 30 mm for azithromycin, and 26 mm for erythromycin; in case of Escherichia coli, were 5 mm for oxytetracycline, 9 mm for amoxicillin, 22 mm for ciprofloxacin, 30 mm for cefotaxime, 31 mm for ceftriaxone, 15 mm for azithromycin, and 0 mm for erythromycin; in case of Salmonella typhi., were 25 mm for oxytetracycline, 24 mm for amoxicillin, 38 mm for ciprofloxacin, 31 mm for cefotaxime, 34 mm for ceftriaxone, 24 mm for azithromycin, and 0 mm for erythromycin. Therefore, ciprofloxacin and ceftriaxone may be the antibiotics of first choice, and cefotaxime and azithromycin may be the second choice among the test antibiotics for the treatment of illness caused by these bacteria.Asian J. Med. Biol. Res. December 2015, 1(3): 457-462

scholarly journals THE OCCURRENCE OF BACTERIAL AGENTS CAUSING MASTITIS IN DAIRY SHEEP AND THEIR RESISTANCE TO ANTIBIOTICS

2020 â—½  
Vol 3 (7) â—½  
Author(s):  
František Zigo â—½  
Katarína Veszelits Lakticová â—½  
Mária Vargová M
Keyword(s):  
Staphylococcus Aureus â—½  
Antibiotic Resistance â—½  
High Resistance â—½  
Subclinical Mastitis â—½  
Causative Organism â—½  
Coagulase Negative Staphylococci â—½  
Isolation And Identification â—½  
Milk Samples â—½  
California Mastitis Test â—½  
Acute Form

Introduction: The aim of this study was to monitor the occurrence of bacterial agents causing mastitis, forms of mastitis and antibiotic resistance in 300 ewes on a farm in Eastern Slovakia. Material and methods: During the milking season, were performed three complex investigations including clinical examination, California Mastitis Test and laboratory analysis of milk samples. The investigations and milk samples were taken in three phases; the beginning (April), the middle (June) and the end (September) of the milking season. Results: Of all the samples (806), 225 (28.0%) were positive for bacterial pathogens. The highest incidence of mastitis (33.3%) was recorded in September, while April (23.8%) and June (25.3%) had lower incidence. The samples from September showed the highest incidence of subclinical mastitis (20.1%), with 13.2% being an acute form of mastitis. Coagulase-negative staphylococci (CNS) were identified in 61.7% of the positive samples. Especially, S. chromogenes, S. epidermidis and S. schleiferi were most frequently isolated.  Staphylococcus aureus was the causative agent in 20.0% of the positive samples and caused acute or subclinical mastitis in the affected ewes. The tested bacteria showed very high resistance to novobiocin (59.5%) and penicillin (51.4%) and high resistance to amoxycillin (35.1%). We found that 80% of Staphylococcus aureus bacteria tested for antibiotic resistance were resistant to novobiocin and 70% were resistant to penicillin. Of all tested CNS, 56.5% were resistant to novobiocin, 39.1% to penicillin, and 34.7% to amoxicillin. Conclusion: Proper isolation and identification of the causative organism play a significant role in the prevention and control of the intramammary infection. In our study, a combinations of Streptomycin, Ciprofloxacin and Tetracycline were the most effective antibiotics for the control of mastitis.

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