Identification of some human pathogenic fungi using four DNA extraction methods

Dermatophytes being animal and human pathogenic fungi infect some human at one point or the other in their lifetime. For effective control of dermatophytes, accurate identification of the specific species/strain involved must be known. Stocks from pathogenic fungi isolated from infected areas on different patients, around Lagos-Nigeria were analysed using molecular methods (DNA extraction, PCR-RFLP and DNA sequencing). Four DNA extraction protocols were employed in the identification of the fungal isolates. Sixteen different fungal isolates were identified, and based on the molecular data these were classified into six species of dermatophytes belonging to the genera Microsporum, Trichophyton and Epidermaphyton, two species of systemic mycoses fungi and eight opportunistic human pathogenic fungi. The results also revealed that CTAB protocol, Modified CTAB protocol and the GNOME kit used in this work were only able to extract non-dermatophytes DNA. Only the Zymo DNA kit was able to isolate dermatophytes DNA. DNA extraction which is the first step in all molecular studies showed that one DNA extraction method might not be able to extract all fungal DNA for proper identification, diagnosis and treatment of fungal infections. Keywords: Dermatophytes, DNA extraction, Identification, Protocols

Production of cellulolytic and xylanolytic enzymes by a phytopathogenic Myrothecium roridum and some avirulent fungal isolates from water hyacinth

The cellulolytic and xylanolytic activity of a pathogenic Myrothecium roridum Tode (IMI 394934) and non-pathogenic Fusarium solani and Curvularia pallescence Boedjin isolates from water hyacinth were investigated. The mycelial plugs of each isolate was grown in submerged cultures of Czapeck Dox broth containing the appropriate carbon source (carboxymethylcellulose, sawdust and homogenized dry water hyacinth leaf) at 25°C for 16 days. The enzyme activity assay was carried out on the culture filtrates obtained. This was measured as micromole sugar released per min. The result obtained showed that the enzyme activity (U/ml) for b-1,4-exoglucanase, b-1,4-endoglucanase and xylanase was maximum 3.70 ± 0.43, 0.95 ± 0.03 and 2.32 ± 0.10, respectively, in C. pallescens Boedjin grown on carboxymethylcellulose and minimum 0.12 ± 0.02, 0.13 ± 0.03 and 0.34 ± 0.01 respectively, in M. roridum grown on homogenized dry water hyacinth leaf. The b-glucosidase activity (U/ml) was highest, 1.74 ± 0.06 in M. roridum grown on sawdust and least, 0.08 ± 0.00 in C. pa llescens Boedjin grown on homogenized water hyacinth leaf broth. The maximum (324.00 ± 19.51 mg/ml) and minimum (130.00 ±5.83 mg/ml) total extracellular protein was produced in M. roridum grown on homogenized dry water hyacinth leaf and carboxymethylcellulose, respectively. This study showed that the phytopathogenic strain of M. roridum is capable of producing cellulases and xylanase enzyme in submerged cultures but to a lesser degree compared to F. solani and C. pallescence Boedjin

Evaluation of acute and subacute toxicity of Alstonia congensis Engler (Apocynaceae) bark and Xylopia aethiopica(Dunal) A. Rich (Annonaceae) fruits mixtures used in the treatment of diabetes

The present study was carried out to evaluate acute and subacute toxicity of a hydroalcoholic extract of a mixture (1:1) of A. congensis bark and X. aethiopica fruits used locally in the treatment of diabetes.Acute toxicity of the extract was evaluated in Swiss albino mice. The animals were fed with the hydroalcoholic extract between the doses of 1.0 to 20.0 g/kg body weight and were observed continuously for the first 4 h and for every hour for the next 24 h, then 6 hourly for 48 h. Wistar rats were also fed with different doses of the extract for 30 days and effects on biochemical parameters evaluated (subacute toxicity model). The LD50 of the extract was found to be above 20.0 g/Kg bodyweight. There was reduction in the plasma glucose and low-density lipoprotein (LDL)-cholesterol levels, and increase in high-density lipoprotein (HDL)-cholesterol level in the treated animals. A significantincrease in the body weight was observed for groups treated with lower doses of the extract while groups treat with higher doses showed no significant weight increase. Aspartate aminotransferases (AST) and alanine aminotransferases (ALT) levels were not affected at lower doses of the extract but there was increase in creatinine levels in all the treated animals. The extract demonstrated good hypoglycaemic effects by lowering the plasma sugar level and also had some beneficial and reduction effects on cardiovascular risk factors. There was no evidence of drug-induced symptoms or death at all the doses of the extract administered in acute study but subacute results revealed a tendency to cause kidney problems on a long-term use

Effect of metabolites produced by Trichoderma species against Ceratocystis paradoxa in culture medium

Metabolites released from Trichoderma viride, T. polysporum, T. hamatum and T. aureoviride were tested in culture medium against Ceratocystis paradoxa, which causes black seed rot in oil palm sprouted seeds. The Trichoderma metabolites had similar fungistatic effects on the growth of C.paradoxa except those from T. aureoviride. The inhibition varied depending on the Trichoderma species producing the metabolites; from 2.0% to 64% in volatile, 0.0% to 74% in non-volatile and 0.0% to 81% from direct-diffusible metabolites. C. paradoxa growth was significantly reduced in the presence ofmetabolites produced by T. viride and T. polysporum than the other species. T. aureoviride had the least growth inhibition, and medium containing direct-diffusible metabolite supported highest inhibition of C. paradox

Physico-chemical properties and heavy metal content of water sources in Ife North Local Government Area of Osun State, Nigeria

The water quality examination in Ife–North Local Government of Osun State Nigeria was conducted by determining the physico–chemical parameters of 40 samples. Surface water, bore holes, wells and pipe borne water samples were collected from major towns in the Local Government Area and analyzed. Results showed temperature range of 26.5 to 33.0°C, pH (6.53 to 8.90), conductivity (63.0 to 1039.0 μS/cm), resistivity (0.00 to 0.02mΩ.cm), salinity (300.00 to 700.00 mg/L), total solids (90.00 to 1175.00 mg/L), total dissolved solids (37.80–622.50mg/l), suspended solids (34.50 to 794.00 mg/L),dissolved oxygen (4.48 to 9.48 mg/L), Biological Oxygen Demand (BOD5) (0.69 to 6.74 mg/L), chloride (3.64 to 184.04mg/L), Nitrate (1.08 to 53.03 mg/L), phosphate (4.99 to 23.07 mg/L) and sulphate (6.02 to 28.95 mg/L). Results of Atomic Absorption spectrophotometric analysis (AAS) of samples for dissolved trace metals; (Cd, Pb, Mn, Zn, Cu, Cr and Fe), showed mean metal concentration to vary widely depending on the source of the water sample as follows: Pb (0.29 ± 0.05 to 6.69 ± 0.53 mg/L), Cd (0.77 ± 0.06 to 2.24 ± 0.08 mg/L), Zn (0.03 ± 2.15 to 0.22 ± 4.64 mg/L), Cu ( 0.18 ± 16.34 to 0.41 ± 5.00 mg/L), Fe (6.00 ± 0.21 to 31.75 ± 0.80mg/L) and Mn (0.14 ± 6.12 to 0.23 ± 99.11 mg/L). The results obtained for the physical parameters agreed with the limits set by both national and international bodies for drinking and domestic water with few exceptions.Key words: Quality, physico-chemical, atomic-absorption-spectrophotometer, domestic, recreational, Ife-North-Local-Government

Evaluation of acute and subacute toxicity of Alstonia congensis Engler (Apocynaceae) bark and Xylopia aethiopica (Dunal) A. Rich (Annonaceae) fruits mixtures used in the treatment of diabetes

The present study was carried out to evaluate acute and subacute toxicity of a hydroalcoholic extract of a mixture (1:1) of A. congensis bark and X. aethiopica fruits used locally in the treatment of diabetes.Acute toxicity of the extract was evaluated in Swiss albino mice. The animals were fed with the hydroalcoholic extract between the doses of 1.0 to 20.0 g/kg body weight and were observed continuously for the first 4 h and for every hour for the next 24 h, then 6 hourly for 48 h. Wistar rats were also fed with different doses of the extract for 30 days and effects on biochemical parameters evaluated (subacute toxicity model). The LD50 of the extract was found to be above 20.0 g/Kg bodyweight. There was reduction in the plasma glucose and low-density lipoprotein (LDL)-cholesterol levels, and increase in high-density lipoprotein (HDL)-cholesterol level in the treated animals. A significantincrease in the body weight was observed for groups treated with lower doses of the extract while groups treat with higher doses showed no significant weight increase. Aspartate aminotransferases(AST) and alanine aminotransferases (ALT) levels were not affected at lower doses of the extract but there was increase in creatinine levels in all the treated animals. The extract demonstrated good hypoglycaemic effects by lowering the plasma sugar level and also had some beneficial and reduction effects on cardiovascular risk factors. There was no evidence of drug-induced symptoms or death at all the doses of the extract administered in acute study but subacute results revealed a tendency to cause kidney problems on a long-term use

Chemical composition, antimicrobial activity, proximate analysis and mineral content of the seed of Detarium senegalense JF Gmelin

Detarium senegalense JF Gmelin (Caesalpiniaceae), commonly known as tallow tree, is used traditionally for the treatment of bronchitis, pneumonia, internal complaints and skin diseases in Tropical Africa. The seed is used as a soup thickener in Eastern Nigeria. Analysis of the petroleum ether extract of the seeds with GC-MS produced ten constituents of which oleic and linoleic acids were the most prominent (30.8 and 44.1% respectively). The extract showed significant antibacterial activity against Staphylococcus aureus, Staphylococcus epidermidis, Micrococcus kristinae, Streptococcus faecalis, Shigella flexneri, Klebsiella pneumonia, Serratia marcescens and antifungal activity against Aspergillus flavus, Aspergillus niger and Penicillium notatum. Proximate analysis revealed that the seeds contain 24.43% carbohydrate, 7.23% protein, 31.16% fiber, 5.89% moisture and 1.93% ash. Mineral content analysis revealed the concentrations of potassium (99.26 mg/g), calcium (71.11mg/g), magnesium (77.83 mg/g), sodium (55.26 mg/g), iron (30.21 mg/g), manganese (7.89 mg/g), zinc (5.26 mg/g) and copper (4.29 mg/g). These results show the nutritional value of the seeds of D. senegalense and justified its use in the traditional treatment of skin diseases.Key words: Detarium, GC-MS, antimicrobial, proximate analysis, mineral content

Prevalence of spontaneous bacterial peritonitis in liver cirrhosis with ascites

Introduction: Spontaneous bacterial peritonitis (SBP) is a common bacterial infection in patients with cirrhosis and ascites requiring prompt recognition and treatment. The aim of this study was to determine the prevalence, and characteristics of SBP among in-patients with cirrhosis and ascites seen at our facility.Methods: Thirty one patients with liver cirrhosis and ascites who were admitted into the Medical ward of the Ekiti State University Teaching Hospital (EKSUTH), Ado-Ekiti, Nigeria from August 2009 to July 2010 were retrospectively studied. All the patients had abdominal paracentesis done within 48 hours of admission under aseptic condition and the data obtained were analyzed. Results: The mean age of the studied population was 62±9 years (age range 43-78 years). Of the 21 that developed SPB, culture positive SBP was present in 66.7% (14/21) while CNNA was found in 33.3% (7/21). The prevalence of MNB was 26% (8/31) in this study. Of those with SBP, 93% had monomicrobial infection with aerobic Gram negative bacilli being responsible in 66.7% of the cases with E.coli (70%) being the predominant organism followed by Klebsiella species. Gram positive organisms accounted for 33.3% with Streptococcal species (60%) being the predominant organism followed by Staphylococcus aureus (40%). Patients with SBP had significantly lower platelet count when compared with those without SBP, p < 0.05. Also, international normalization ratio (INR) was significantly higher in those patients with SBP compared with those without SBP, p < 0.05. The poor prognostic indicators found in this study were; low ascitic protein, hepatic encephalopathy, coagulopathy, renal dysfunction (creatinine >2mg/dl) and leukocytosis (p < 0.05). Conclusion: It is therefore imperative to do diagnostic abdominal paracentesis for cell count and culture in any patient with onset of ascites or cirrhotic patients with ascites and suggestive symptoms compatible or suggestive of SBP.Key words: Spontaneous bacterial peritonitis, abdominal paracentesis, liver cirrhosis, ascite

Some haematological and biochemical profile of blood of Nile tilapia (Oreochromis niloticus) fed on diets containing watermelon (Citrullus lanatus) seedmeal

The haematological and biochemical profile of blood of Nile tilapia (Oreochromis niloticus) fed on diets  containing watermelon (Citrullus lanatus) seedmeal were evaluated using packed cell volume (PCV);  haemoglobin content (Hb), white blood cell count (WBC), red blood cell count (RBC), mean corpuscular  haemoglobin (MCH), mean corpuscular volume (MCV), mean corpuscular haemoglobin concentration (MCHC), blood glucose, cholesterol, total protein, albumin and globulin as indices. 150 tilapia fingerlings of average  weight 6.12±0.05g were acclimatized for a week, weighed and allotted into five dietary treatments; DT1, DT2, DT3, DT4 and DT5 containing 0, 15, 30, 45 and 60% Citrullus lanatus replacement levels with soybean meal, respectively. The diets were isonitrogenous and isolipidic. Each treatment was replicated three times with ten fish per replicate. Fish were fed 5% body weight on two equal proportion per day. The results from the study indicated that there was no significant difference (p>0.05) in the haematological and biochemical parameters of the blood of fish fed on the various dietary treatments.Keywords: Blood biochemistry, Citrullus lanatus, haematology, tilapia, watermelo

Olax Subscorpioidea Oliv. (Olacaceae): An Ethnomedicinal and Pharmacological Review

Background: Olax subscorpioideaOliv. (Olacaceae) is a woody shrub that is widely distributed in Africa. It hastrado-medicinal importance and is used in the treatment of asthma, cancer, convulsion, diabetes, intestinal worm infections, jaundice, mental illnesses, neurodegenerative disorders, sexually transmitted infections, swellings and rheumatism, and yellow fever. Aims: To review available literature on the phytochemistry, ethnobotany, pharmacology and toxicity of Olax subscorpioideaOliv. Method:Published findings were searched in online databases such as Web of Science, Scopus, Pubmed, Google Scholar and other relevant sources, and the data were sorted by relevance. Combinations of keywords used in the search include Olax subscorpioidea, Olacaceae, Olax, Ewe Ifon, and African medicinal plants. Results: The presence ofalkaloids, anthraquinones, cardiac glycosides, flavonoids, phenoliccompounds, proanthocyanidins, saponins, tannins and triterpeneshasbeen reported from O. subscorpioidea. Several secondary metabolites have been identified, importantly the cytotoxic santalbic acid from the seeds. Bioactivity studies on this plant demonstrated its medicinal potential mainly asan analgesic, anthelmintic, anti-arthritic, antidepressant, antihyperglycaemic, anti-inflammatory, antioxidant, antimalarial and antimicrobial agent. Oral acute toxicity of the leaf extracts in rats appears to be negligible. Conclusion: Published literature available to date onO. subscorpioideaOliv. provides some preliminary scientific basis for the ethnomedicinal uses of this plant. However, some ethnomedicinal uses have not been scientifically, only a limited amount of information is available on properly isolated and identified phytochemicals from this plant that link to its bioactivities