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Detection of methicillin resistant and slime factor production of coagulase negative Staphylococcus spp. in bovine clinical mastitis by using PCR

Research Article of American Journal of Microbiology and Immunology Detection of methicillin resistant and slime factor production of coagulase negative Staphylococcus spp. in bovine clinical mastitis by using PCR S. M. El-Berbawy*; S.M. Sayed*;El-Toukhy, E. I. ** and Amal, A. Megahed*** * Assiut Lab. (Bacteriology Department) ** AHRI (Biotechnology Depart.) Dokki *** Port Said Lab. (Bacteriology Depart.) This study aims to investigate the slime production of Coagulase negative staphylococci (CoNS) isolates by phenotypic method on Congo Red Agar plates (CRA) and Genotypic detection of icaA, icaD and mecA genes by polymerase chain reaction (PCR). Out of 105 milk samples obtained from clinical bovine mastitis, 101samples (96.2%) were positive for bacterial growth. CoNS isolates was detected in 20 isolates with a percentage of 19.8%. Their ability to form biofilm as one of the most important virulence factors of the organisms using Congo Red Agar (CRA) method was investigated in which 13 out of 17 CoNS isolates (76.47%) were found to be slime producers. By PCR, mecA gene was found in threeout of 6 CoNS isolates (50%). Also six (100%) and three (50%) isolates were positive for icaA gene and icaD gene, respectively. In addition one isolate out of the six CoNS isolates (16.67%) was positive for the presence of icaA, icaD and mecA genes and also has the ability to form biofilm. The in vitro activities of CoNS against 11 selected antimicrobial agents referred that the highest resistance rate of CoNS observed to Lincomycin (100%), followed by Cefotaxime (94.41%), Oxacillin (58.82%), Ampicillin (47.06%) and Penicillin (41.18%), while the highest rate of sensitivity observed to Enrofloxacin and Gentamicin (100%, for each), followed by Doxycycline (94.11%).Conclusion, the findings of the present study demonstrated the ability of CoNS isolated from bovine clinical mastitis to form biofilms. This must be considered as an alarming situation, ...

The Accumulation Of Cadmium In Corn At Different Levels Of Soil Ph

Research Article of American Journal of Microbiology and Immunology The Accumulation Of Cadmium In Corn (Zea Mays L.) At Different Levels Of Soil Ph Rabah S. Shareef*1,2, Awang Soh Mamat*3, Zakaria Wahab*4, Ibni Hajar Rukunudin*5 1 College of Education-Al-Qiam, University of Anbar, Iraq. 2 School of Bioprocess Engineering, University Malaysia Perlis, Perlis, Malaysia. 3 School of Bioprocess Engineering, University Malaysia Perlis, Perlis, Malaysia. 4 School of Bioprocess Engineering, University Malaysia Perlis, Perlis, Malaysia. 5 School of Bioprocess Engineering, University Malaysia Perlis, Perlis, Malaysia. A pot experiment was carried out in the plastic greenhouse at the Institute of Sustainable Agrotechnology (INSAT), University Malaysia Perlis in Sungai Chuchuh, Perlis, Malaysia. During season of the cultivation of 2014 investigate the impact of soil pH and cadmium on growth of corn plant (Zea mays L.). Twenty five were arranged in factorial experiments according to the Complete Randomized Design (CRD), with three replicates. Five levels of soil pH were, pH4, pH5.2 (i.e., the original value), pH6, pH7 and pH8 and five levels of cadmium (Cd); Cd 1, Cd 2, Cd 3, Cd 4 and Cd 0 where the amounts (2, 4, 6, 8 mg. kg-1 soil and control treatment without add cadmium) are applied as CdCl2. Thus, the total numbers of pots were 75 pots. The results of this investigation revealed that; The decrease of soil pH led to significant effect to increase concentration of cadmium in root, stem, leaves and grain of corn, where the level of pH 5.2 gave highest concentrations of Cd compared with pH 8. the interactions between soil pH and cadmium led to increase of reduced the accumulation of cadmium in the corn, where it gave the interaction between pH 8 and Cd 0 lowest concentration of cadmium in root, stem, leaves and grain. Keywords: Accumulation, Cadmium, Corn, Soil pH, ...

A contribution on Coliforms causing mastitis in cows with reference to serotypes and virulence factors of E. coli isolates

Research Article of American Journal of Microbiology and Immunology A contribution on Coliforms causing mastitis in cows with reference to serotypes and virulence factors of E. coli isolates Sayed, S. M. Egypt- Animal Health Research Institute (Assiut Regional Lab., Bacteriology Dept.) Escherichia coli (E. coli) is the predominant coliform species causing intramammary infections. Where in the present study, E. coli isolates were 18 strains (17.82%) followed by Enterobacter aerogenes 3 strains (2.97%) and Klebsiella pneumoniae one strain (0.99%) from 101 clinical mastitic milk samples of cows. Eighteen E. coli isolates were serotyped to nine different serogroups; O111:H4 (3), O127:H6 (3), O26 (2), O126 (2), O119:H6 (1), O114:H21 (1), O55:H7 (1), O44:H18 (1), O124 (1) and (3) untyped. Virulence tests were performed on the 18 isolated E. coli, it was found that 15 isolates (83.3%) were serum resistant, 13 isolates (72.2%) had Congo Red binding activity, 6 isolates (33.3%) were invasive and one isolate (5.6%) had haemolytic activity. PCR was applied to detect the presence of Shiga like toxin producing E. coli (stx1 and stx2 genes) on the nine different strains (one strain for each serogroup), where stx1 and stx2 were found in 8 (88.9%) and 4 (44.4%) of the nine examined strains, respectively. While stx1 and stx2 genes were found together in 3 strains (33.3%). Conclusions:  E. coli isolates usually posses one or more virulence factors that may help in establishment at the infection site and subsequently causing clinical bovine mastitis. Keywords: Coliforms, E. coli, serotypes, virulence factors, stx1 and stx 2 ...

Bacteriological study on staphylococcal bovine clinical mastitis with reference to methicillin-resistant Staph. aureus (MRSA)

Research Article of American Journal of Microbiology and Immunology Bacteriological study on staphylococcal bovine clinical mastitis with reference to methicillin-resistant Staph. aureus (MRSA) Sayed, S. M. Egypt- Animal Health Research Institute (Assiut Regional Lab., Bacteriology Dept.) This descriptive study was done on 101 milk samples obtained from clinically mastitic dairy cows in Assiut Governorate, Egypt. Staphylococcus aureus (S. aureus) was the main causative agent of clinical mastitis (34.65%) followed by S. saprophyticus (10.89%), S. intermedius and S. epidermidis (8.91%, for each). The other causative agents (non Staph. Spp.) were identified. Sensitivity test of S. aureus isolates was performed against 11 antimicrobial agents, where found that 21 S. aureus strains (60%) were methicillin resistant S. aureus (MRSA). Ten MRSA strains were subjected for: I- slime-producing factor on Congo Red Agar (CRA) plates phenotypically, as 6 isolates (60%) were positive for slime production. II- PCR which was optimized targeting mecA, icaA and icaD genes, where 5 isolates (50%) were positive for mecA gene. Six isolates (60%) and 8 isolates (80%) were positive for icaA and icaD genes, respectively. Five strains (50%) were positive for both icaA and icaD genes. Also 3 strains (30%) were positive for all mecA, icaA and icaD genes. Conclusion, it was concluded that bovine staphylococcal mastitis was the most predominant issue where S. aureus was the main cause. Detection of mecA gene in S. aureus isolates indicating that several cases suffering from S. aureus mastitis have an MRSA problem. Genotypic determination of mecA gene proved the most reliable method for detection of methicillin-resistant S. aureus. The present work paid an attention to the 3 MRSA strains (30%) were positive to all tested genes rather than slime production as the worst isolated strains all over this study (multidrug resistant, slime producing as well as carrying mecA, icaA and icaD genes). In vitro Enrofloxacin, Gentamicin and Doxycycline the most effective drugs for Staph. spp. clinical mastitis and should be recommended for treatment of such cases of bovine mastitis.    Keywords: Cows, clinical mastitis, methicillin-resistant Staphylococcus aureus, ...

Antimicrobial activity of crude leaf extracts against Enterococcus faecalis

Research Article of International Journal of Traditional and Complementary Medicine Antimicrobial activity of crude leaf extracts from medicinal plants against Enterococcus faecalis Rachuonyo HO1* Gatheri GW2 Nyamache AK1 1Department of Microbiology, Kenyatta University 2Department of Plant sciences, Kenyatta University Review method: Single-blind; Peer reviewer comments: 1. Enterococcus faecalis is a Gram negative, commensal enteric bacterial pathogen usually located in the intestinal tracts of either animals or human beings. The pathogenic members of enteric bacteria’s are usually associated to infections that are characterized by enteric fevers, abdominal pain, and diarrhoea and vomiting. This study aimed at determining the effects of the selected medicinal plants extracts against Clinical isolate of Enterococcus faecalis obtained from Kenyatta University health Centre. Kirby Bauer method was used to determine the antimicrobial activity of the plants extracts against Enterococcus faecalis. All the plant extracts showed antimicrobial activity against Enterococcus faecalis with Tagetes minuta producing the largest average zones of inhibition of 18.67±1.03mm when compared to the other plant extracts. Vernonia lasiopus was more active at low concentrations (MIC 3.9µg/ml; MBC 5.0µg/ml) while Aloe secundiflora was less active (MIC 7.0µg/ml; 9.7µg/ml). Ciprofloxacin (5µg/ml) was used as a positive control producing an average zone of inhibition of 21.67±2.66mm while Methanol (33.50±2.56mm) and 4% Dimethyl sulphoxide (0.00±0.00mm) were used as negative control. Qualitative phytochemical screening showed the presence of four phytochemicals namely; saponins, tannins, alkaloids and flavonoids. The study provides insight about the antimicrobial activity of the plant extracts and if they can be used in treatment of as an antimicrobial agent against infections caused by Enterococcus faecalis. Keywords: Qualitative phytochemical screening; Antimicrobial agents; Enterococcus faecalis; Ciprofloxacin; Clinical isolate ...

Different Levels of Nitrogenous Fertilizer Application for Growth, Flowering and Yield Attributes of Glori lily (Gloriosa superba L.)

Research Article of Journal of Herbal Medicine Research Different Levels of Nitrogenous Fertilizer Application for Growth, Flowering and Yield Attributes of Glori lily (Gloriosa superba L.) S. Deivasigamani Department of Agronomy, PGP College of Agricultural sciences, (Affiliated to Tamilnadu Agricultural University, Coimbatore-3)  Palani Nagar, Namakkal, Tamilmadu, India Review method: Single-blind; Peer reviewer comments: 1. Field experiment were conducted at farmer’s field, Devanur village, Ariyalur District of Tamil Nadu during 2008 to study the Different Levels of Nitrogenous Fertilizer Application for Growth, Flowering and Yield Attributes of glori lily. There were Five different treatments viz., Control  (T0), 100 per cent recommended  N (T1), 75 per cent recommended N (T2), 50 per cent recommended N (T3) and 25 per cent  recommended N (T4). The treatments were replicated four adopting RBD design. The growth and yield attributes of glori lily viz., plant height, leaf area index, number of primary branches plant-1, number of secondary branches plant-1, number of tertiary branches plant1,drymatter production (kg ha-1), days to fifty per cent flowering, number of pods plant-1, number of seeds pod-1 and hundred seed weight (g), were favorably  increased by the application of 100 per cent  recommended N (T1).  Seed and tuber yield (Kg ha-1) were greatly increased and the maximum seed and tuber yield were recorded. The uptake of NPK by glori lily was significantly increased by the above treatment combination. Application of 100 per cent recommended N (120 kg ha-1) (T1) gave the highest Gross income, Net income and return per rupee invested. Based on the above results, it can be concluded that inorganic use of 100 per cent recommended N (120 kg ha-1) is found to be an appropriate agro-technique for augmenting the productivity and profitability of glori lily without altering the soil fertility. It is a basic constituent in all proteins, enzymes, chlorophyll, certain parts ...