Combination of essential oil and ciprofloxacin to inhibit/eradicate biofilms in multidrug-resistant Klebsiella pneumoniae.

AIM: This study aimed to test biofilm inhibition activities of each of essential oils (EOs), main compounds of EOs and enzymes against pathogenic Klebsiella pneumoniae. METHODS AND RESULTS: The effect of seven EOs and three enzymes was tested on formation and eradication of K. pneumoniae biofilm. Peppermint oil showed a robust biofilm inhibitory effect, causing inhibition that ranged from 69·2 to 98·2% at 5 µl ml-1 . Thyme oil was found to have the best biofilm eradication ability, causing eradication that ranged from 80·1 to 98·0% at 10 µl ml-1 . The most effective EOs were analysed by GC/MS, to determine the major chemical constitutes of each oil. Pure menthol was found to cause 75·3-97·5% biofilm inhibition at 2·5 µg ml-1 , whereas thymol caused 85·1-97·8% biofilm eradication at 5 µg ml-1 . However, moderate inhibition activity was detected for α-amylase and bromelain, while poor activity was detected for ß-amylase. Ciprofloxacin combination with thyme oil and thymol was found to enhance antibiotic activity, and affect biofilm cell viability. The observed inhibitory/eradication activity on K. pneumoniae biofilms was confirmed by scanning electron microscopy. CONCLUSIONS: Thyme and peppermint EOs, and their active components are promising antibiofilm agents alone and/or in combination with ciprofloxacin to inhibit/eradicate biofilms of K. pneumoniae. SIGNIFICANCE AND IMPACT OF THE STUDY: The presented results suggest the potential application of EOs against infections, caused by biofilm-producing K. pneumoniae, to prevent biofilm formation or decrease their resistance threshold. Moreover, the combination of EOs with ciprofloxacin minimizes the antibiotic concentration used and accordingly the potential accompanying toxic side effects.

The first use of EaeI restriction enzyme in DNA diagnosis of Hb Q-India.

INTRODUCTION: The α-chain variant Hb Q-India (c.193G>C) is caused by a point mutation GAC→CAC at codon 64 of the α1 globin gene and is clinically silent. Point mutations can be diagnosed easily by many simple polymerase chain reaction (PCR) techniques including PCR-restriction digest, but for Hb Q-India the restriction digest has never been described. In this work we aimed to develop a restriction enzyme digestion assay for DNA diagnosis of Hb Q-India, in order to increase the panel of restriction enzymes used in DNA diagnosis of haemoglobinopathies and also as a simple cheap alternative to the ARMS-PCR method. METHODS: A restriction enzyme digestion assay was designed for diagnosis of Hb Q-India using the restriction enzyme EaeI enzyme as the Hb Q-India mutation abolishes the recognition site of this enzyme. Patients were screened for an abnormal haemoglobin by high performance liquid chromatography (HPLC) and those had an abnormal peak with a retention time between 4.7 and 4.8 minutes were selected for diagnosis at the molecular level. The α1 globin gene was amplified in 12 cases with a presumed diagnosis of Hb Q-India by HPLC and isoelectric focusing (IEF), and the amplified products were subjected to the EaeI digestion. RESULTS: All the 12 cases were diagnosed positive (100%) for Hb Q-India by the EaeI restriction enzyme digest. They were heterozygotes for the mutation. CONCLUSION: EaeI restriction enzyme digestion can be used as a simple and robust alternative method to ARMS-PCR for DNA diagnosis of Hb Q-India. The EaeI restriction enzyme can be added to the panel of restriction enzymes used in the DNA diagnosis of the abnormal Hb variants. Concomitant use of HPLC and IEF can be used efficiently for presumed diagnosis of this rare variant.

Haemoglobin (Hb) G-Philadelphia, Hb Stanleyville-II, Hb G-Norfolk, Hb Matsue-Oki and Hb Mizushi can form a panel of α-chain variants that overlap in their phenotype: the novel use of StyI to screen for Hb G-Philadelphia.

INTRODUCTION: Haemoglobin (Hb) G-Philadelphia mutation is a common alpha-globin chain variant [α68(E17)Asn > Lys]. Combined high performance liquid chromatography (HPLC) and isoelectric focusing (IEF) can be used in a presumptive diagnosis of Hb G-Philadelphia, but there are other α-chain variants with a similar phenotype that cannot be excluded. Our aim was to develop a novel StyI restriction enzyme assay to diagnose the common Hb G-Philadelphia mutation and to identify any other variants with a similar phenotype by DNA sequencing. METHODS: Thirty-one cases given a presumptive diagnosis as Hb G-Philadelphia by HPLC and IEF were subjected to DNA analysis by restriction enzyme digestion using StyI. Negative cases were then subjected to DNA sequencing. RESULTS: Twenty-two cases (78.6%) of 28 cases amplified were tested positive for Hb G-Philadelphia by StyI restriction digestion. Sequencing of the six negative cases revealed two cases of Hb G-Philadelphia with C→A mutation in codon 68 in α2 globin gene, plus one case each of Hb G-Norfolk Hb Stanleyville-II, Hb Matsue-Oki and Hb Mizushi. CONCLUSION: A novel StyI restriction enzyme can be used to confirm the commonest type of Hb G-Philadelphia. DNA sequencing identified four other α-chain variants with a similar HPLC and IEF phenotype.

Removal of chromium(III) from tannery wastewater using activated carbon from sugar industrial waste.

Chromium is commonly found in huge quantities in tannery wastewaters. For this reason, the removal and recovery of the chromium content of tannery wastewaters is crucial for environmental protection and economic reasons. Removal and recovery of chromium were carried out by using low-cost potential adsorbents. For this purpose three types of activated carbon; C1, the waste generated from sugar industry as waste products and the others (C2, C3) are commercial granular activated carbon, were used. The adsorption process and extent of adsorption are dependent on the physical and chemical characteristics of the adsorbent, adsorbate and experimental condition. The effect of pH, particle size and different adsorbent on the adsorption isotherm of Cr(III) was studied in batch system. The sorption data fitted well with Langmuir adsorption model. The efficiencies of activated carbon for the removal of Cr(III) were found to be 98.86, 98.6 and 93 % for C1, C2 and C3, respectively. The order of selectivity is C1>C2>C3 for removal of Cr(III) from tannery wastewater. Carbon "C1" of the highest surface area (520.66 m(2)/g) and calcium content (333.3 mg/l) has the highest adsorptive capacity for removal of Cr(III). The results revealed that the trivalent chromium is significantly adsorbed on activated carbon collected from sugar industry as waste products and the method could be used economically as an efficient technique for removal of Cr(III) and purification of tannery wastewaters.

Anaerobic fermentation of gelatinized sago starch-derived sugars to acetone-1-butanol-ethanol solvent by Clostridium acetobutylicum.

A study of the kinetics and performance of solvent-yielding batch fermentation of individual sugars and their mixture derived from enzymic hydrolysis of sago starch by Clostridium acetobutylicum showed that the use of 30 g/L gelatinized sago starch as the sole carbon source produced 11.2 g/L total solvent, i.e. 1.5-2 times more than with pure maltose or glucose used as carbon sources. Enzymic pretreatment of gelatinized sago starch yielding maltose and glucose hydrolyzates prior to the fermentation did not improve solvent production as compared to direct fermentation of gelatinized sago starch. The solvent yield of direct gelatinized sago starch fermentation depended on the activity and stability of amylolytic enzymes produced during the fermentation. The pH optima for alpha-amylase and glucoamylase were found to be at 5.3 and 4.0-4.4, respectively. alpha-Amylase showed a broad pH stability profile, retaining more than 80% of its maximum activity at pH 3.0-8.0 after a 1-d incubation at 37 degrees C. Since C. acetobutylicum alpha-amylase has a high activity and stability at low pH, this strain can potentially be employed in a one-step direct solvent-yielding fermentation of sago starch. However, the C. acetobutylicum glucoamylase was only stable at pH 4-5, maintaining more than 90% of its maximum activity after a 1-d incubation at 37 degrees C.

Toxoplasma infection of stray cats in Egypt.

A seroepidemiological survey of cats for Toxoplasma antibody revealed a high infection rate. No significant difference was detected between males and females or between rural and urban areas, although acute infection seems to be more prevalent in males and in rural areas as revealed by the high titres of the dye test. Toxoplasmosis in cats, as revealed by serologic surveys, seems to prevail more during the warm rather than the hot season of the year being favoured by milder temperature and higher relative humidity and rainfall in the former season. Futhermore, it was concluded that infection rate with Toxoplasma increases with the age of the host. Toxoplasma antibody in the young age groups is to a great extent of maternal origin. Study of the relationship of Toxoplasma to Isospora showed no cross immunity. They follow a reversed pattern with increasing age of the host. Data also shows the high specificity of dye testing for detection of Toxoplasma antibody. Feeding experiments show that a relatively high percentage of cats shed oocysts in their faeces. Not only seropositive but also seronegative cats excrete oocysts, though at a lower rate in the latter case. The majority of cats shedding oocysts are those with either low positive titres or seronegative cats.