Gluten sensitivity is one of the prominent features of celiac disease (CD) which is an autoimmune disorder characterized by damaged lining of the small intestine. CD was known already to ancient Greeks as κοιλιακÏŒς (keeleeakoss) i.e. disease of the abdominal cavity hence celiac. Focus of this Commentary article is on rather complex definition of CD and its emerging new forms the example of which is non-celiac gluten sensitivity. It is becoming evident that to formulate more effective treatments, these associations and newly identified disease entities deserve attention from both academic and clinical communities.
Aims: Beta-lactamase production and subsequent resistance to β-lactam drugs has been a global concern in the treatment of Gram negative anaerobes. The aim of this study was to identify F. nucleatum strains producing Class D β-lactamase through the detection of FUS-1 (OXA-85) resistance gene. Place and Duration of Study: Department of Preventive Dentistry, Lagos University Teaching Hospital, Idi-Araba, between February 2010 and November 2010. Methodology: Twenty two oral clinical samples were obtained from patients with chronic periodontitis who admitted to previous use of amoxicillin. Antibacterial susceptibility of the bacterial isolates was determined by E-test on Brucella Blood agar. Amplification of the bacterial DNA was carried out by PCR using F. nucleatum species-specific primer, FUS-1 specific for blaFUS-1 and strain-specific primers for subspecies nucleatum,, fusiforme, polymorphum and vincentii. Results: From the 19 samples collected, F. nucleatum was isolated, and the identity of the isolates was confirmed by PCR. Four of the isolates produced similar bands with the control strain, 3 (15.7%) strains were able to produce amplication with FUS-1 primer specific for blaFUS-1 gene found in β-lactamase producing F. nucleatum subsp. polymorphum. Conclusion: This study shows the presence of class D β-lactamase producing F. nucleatum species in Nigeria.
Aims: Untreated wastewater is usually used for crop irrigation in developing countries; however it contains a lot of pathogenic bacteria. This study was carried out to determine the fate of E. coli contained in wastewater in a hydromorphic soil. Study Design: Environmental microbiology Place and Duration of the Study: This study was carried out in the experimental field of the Dschang University, during the dry season (November 2011- Mars 2012) and the rainy season (June 2012-September 2012). Methodology: Six plots of 4 m2 each were tilled in 400 m2 surface area in the dry and in the rainy seasons. Wastewater was collected from the experimental wastewater treatment station in the University of Dschang; it was applied on three plots, and three other plots were used as controls. Once every week, soil samples were taken on the surface (0 - 10 cm), in the medium (20 - 30 cm) and at the water table level (40 - 50 cm). Levels of E. coli in soil samples were determined on “Lactose Tergitol® 7 Agar with TTC” medium, and biochemical confirmation tests were carried out (tests of indol, Simmons citrate, gas production, mobility, fermentation of mannitol, glucose and lactose). Results: In the rainy season, E. coli was detected on the soil surface until the 112th day, while in the dry season detection did not exceed the 63rd day. E. coli was detected in the deeper layers of the soil (20 - 30 and 40 - 50 cm) from the 14th and the 70th day respectively. This helps to estimate the speed of vertical migration to be between 5 and 18 mm per day. Conclusion:E. coli bacteria contained in urban wastewater survive for a long-time in hydromorphic soils and reach significant depths, and can consequently pose serious problems of public health.
Aim: This study was undertaken to determine the prevalence of Salmonella spp contamination in the Jamaican poultry industry and its environments. Materials and Methods: A total of 45 farms across 6 Jamaican parishes were selected for this study. A total of 6693 specimens from animals and the environment were investigated for the presence of Salmonella spp. All specimens were placed in an igloo with ice packs and transported to the laboratory for analysis. Bacteriological media obtained from Difco Laboratories Detroit MI U.S.A were used for the isolation and identification of Salmonella spp. Salmonella serological typing was performed to determine the Salmonella serovar by standard procedures. Results: This study revealed a low prevalence of Salmonella contamination/infection in both small and large entities in the poultry industry in Jamaica. The overall prevalence was 1 % (79 positive out of 6693 specimens). However, a higher prevalence of Salmonella was observed in the case of those operations which practiced “organic” poultry farming. It was shown that two Salmonella serovars including Augustenborg and Kentucky, identified during the study, are newly reported serovars in Jamaica. The sources of Salmonella infection varied from poultry itself to other species, such as rodents, pigs and insects. Improper disposal of broken eggs, wet bedding and other fomites contributed to Salmonella contamination. Conclusions: The results of the study indicate possibility of salmonellosis (zoonosis) in Jamaica, although the prevalence of Salmonella spp was low, and the need for improved quality of the food industry, animal care and human health to prevent salmonellosis.
Aims: To identify and determine the bacteria associated with skin and soft tissue conditions of fungal infections. Place and Duration of Study: Sample area was Plateau State Nigeria and sample collection and analysis was done in Dermatophilosis Research Centre, National Veterinary Research Institute Vom, Plateau State, Nigeria, between September 2011 and December 2012. Methodology: Nine hundred and forty (940) human skin and nail scraping samples from different parts of the body were collected from subjects referred to the Centre from different hospitals with visible skin infections. Sample analysis were carried out using standard microbiological methods which include: Wet mount, tease mount, culture and biochemical tests were used to process and analyze for the isolation and identification of fungi and bacteria. Results: Out of 940 samples, 892(94.9%) yielded fungal species which include: Microsporum 45(4.8%), Trichophyton 176(18.7%), Aspergillus 216(22.9%), Epidermophyton 32(3.4%), Candida 72(7.7%), Mucor 141(15.0%), Rhizopus 52(5.5%), Fusarium 12(1.3%), Bipolaris 23(2.5%), Sporothrix 74(7.9%), Penicillium 32(3.4%) and Curvularia 17(1.8%). All samples 940 (100%) yielded an array of bacteria which include: Staphylococcus aureus 125(13.3%), Staphylococcus epidermidis 145(15.8%), Micrococcus luteus 233(24.8%), α-hemolytic Streptococci 89(9.5%), Escherichia coli 59(6.3%), Proteus mirabilis 113(12%), Bacillus subtilis 78(8.3%) and Klebsiella pneumonia 98(10.4%). Staphylococcus aureus, Staphylococcus epidermidis and Micrococcus luteus were isolated from all sites of infection while Micrococcus luteus was isolated from all moist ulcerous and dry scaly skin infections. Conclusion: This study showed the presence of bacteria in high frequency in and around skin and soft tissue infection sites on the body. Micrococcus luteus was the most prevalent bacterial organism associated with skin and soft tissue conditions of fungal infections. Under favourable conditions, some of the bacteria isolated can establish infections through broken skin hence complicating or prolonging treatment of the skin infection.
Alpha amylase is an important enzyme used in different industries, which degrades starch into smaller disaccharides. Extracellular α-amylase producing organisms were isolated from soil samples from Mauritius and identified by standard biochemical tests. In this study, the high yielding strain was used for amylase production. The potential of four readily available substrates, namely sugarcane bagasse, potato peel, kitchen wastes and banana peel to induce amylase production was investigated. Different parameters like temperature (30ºC, 40ºC, 50ºC, 60ºC & 70ºC), different pH (5.0, 6.0, 7.0, 8.0 & 9.0) and inoculum sizes (10%, 20%, 50%, 100% & 150% v/w) were used for the α-amylase production. It was found that α-amylase production and activity was highest for potato peel at 50ºC at pH 6.0 and inoculum size 50% (v/w). Amylase assays performed at different incubation temperatures (30ºC - 60ºC) and pH (5-9) showed that the amylase worked best at 50ºC and pH 7.Based on results of biochemical tests and 16S ribosomal RNA gene sequences, the isolate was identified to belong to the Betaproteobacteria, closely related to Naxibacter haematophilus (99% sequence similarity to the type strain).
Aims: To determine the antiplasmodial activity of methanolic extract of T. avicennioides and its effect on oxidative stress and the lipid profiles in mice infected with Plasmodium berghei. Study Design: Mice used for this study were grouped into five. The first group was not infected with malaria parasite (normal control), the second group was infected with the parasite but not treated with antimalarial drugs (negative control), the third group was infected with the parasite and treated with 5mg/kg body weight of artesunat (positive control), while the fourth and fifth groups were infected with malaria parasite and treated with 100 and 200mg/kg of T. avicennioides respectively. Methodology: The parasitaemia was monitored for five days. The animals were sacrificed on the fifth day and the blood was collected. The serum was used to assess the biochemical parameters using randox kits. Results: While parasite density increases in the negative control per day, there was reduction in parasite density in treated groups. The parasite clearance was significantly higher (P = .05) in those treated with 200mg/kg of T. avicennioides than those treated with 100mg/kg of T. avicennioides and 5mg/kg of artesunat. The malondialdehyde level was significantly higher in the negative control, while superoxide dismutase and catalase levels were significantly reduced when compared with group treated with 200mg/kgbdwt of T. avicennioides. HDL level was significantly higher (P = .05) in those treated with 200mg/kg than in the normal, negative and positive control. The triglycerides level was significantly higher in the negative control when compared with the group treated with the extract of T. avicennioides. Conclusion: This study showed that the methanolic extract of T. avicennioides display dose-related in vivo antiplasmodial and antioxidant activities as well as reduced the serum and liver lipoprotein cholesterol in mice infected with P. berghei.
Aims: To investigate the degradation of 4-chlorophenol by a moderately halophilic bacterial consortium and to study the effect of nitrogen sources on the degradation of 4- chlorophenol. Study Design: 4-chlorophenol degradation by moderate halophiles. Place and Duration of Study: Centre for Environmental studies, Microbiology Lab, Anna University, Chennai, Tamil Nadu, between Jan 2009 and May 2009. Methodology: 4-chlorophenol was degraded with the moderately halophilic bacterial consortium in the minerals salts medium and degradation was analysed by gas chromatography. Effect of NaCl concentrations and alternate nitrogen sources on the degradation of 4-chlorophenol was determined. Results: The bacterial consortium was able to degrade 4-Chlorophenol at a range of NaCl concentrations, where the optimum degradation was obtained at 50 g/L of NaCl. Addition of alternate nitrogen sources like tryptone and urea did not enhance the degradation of 4-Chlorophenol. Conclusion: Bacterial consortium degraded 4-chlorophenol at optimum NaCl concentration of 50 g/L of NaCl. Addition of yeast extract as nitrogen source showed higher degradation than the nitrogen sources.
Aims: To determine the prevalence of acquired pAmpCs in clinically important and relevant enterobacterial species and to characterize the molecular types of pAmpC present in our geographic area. Methodology: Sixty Enterobacterial clinical isolates resistant to third generation cephalosporins and to cephamycins were included in the study. Samples were collected for a period of 6 months between July 2008 and December 2008 from Theodor Bilharz Research Institute (TBRI), Egypt. Bacterial species were identified using API E20. AmpC genes clusters: (bla ACC, bla EBC, bla FOX, bla CMY, bla MOX, and bla DHA) were tested by PCR and DNA sequencing. Clonal relatedness of AmpC-producing Klebsiellae isolates was determined by Pulsed Field Gel Electrophoresis (PFGE). Results: AmpC genes were detected in 28.3% (17/60) of the study population including E. coli, Klebsiella and Proteus mirabilis(P mirabilis). CMY-2 enzyme was found disseminating in all 6 AmpC-positive Escherichia coli(E. coli) and in 6/10 of Klebsiellae species. Only one Klebsiella pneumonia (K. pneumonia) isolate harbored CMY-4 while DHA-1 was detected in 3 Klebsiellae and in one P. mirabilis isolate. PFGE patterns showed no clonal relatedness among the 6 CMY-2-positive Klebsiella isolates. Conclusion: Plasmid-mediated AmpC enzymes are important mechanisms of resistance to ß- lactam drugs. CMY-2 and DHA-1 are the most common gene clusters of pAmpC in our region. AmpC-type resistance in our hospital setting is not due to the dissemination of clonal strains but due to the spread of resistant genes. This is the first report from Egypt identifying DHA-1 and CMY-4 in enterobacterial isolates.
Aims: The aim of this study was to investigate the prevalence of diarrhoea causing human pathogen V. cholerae and other vibrios from different environmental and seafood samples in Tamil Nadu, India. Place and Duration of Study: Laboratory of Clinical Microbiology, Department of Bio-Medical Science, School of Basic Medical Sciences, Bharathidasan University, Tiruchirappalli, Tamil Nadu, India between 2012 and 2013. Methodology: Seafood, water and plankton samples were collected at different locations of Tamil Nadu, India. All the samples were primarily enriched with alkaline peptone water (APW). 2-3 loopful of overnight cultures were streaked onto Thiosulphate Citrate Bile salt Sucrose (TCBS) agar plates. Suspected Vibrio cholerae, V. parahaemolyticus and other vibrios were picked up and identified by using standard biochemical and serological characterization and also by molecular methods. Results: Among the various samples that includes freshwater, coastal water, plankton and various seafoods, only plankton and seafood samples were found to be harbored with V. cholerae, V. parahaemolyticus and V. fluvialis. The remaining samples were negative for vibrios. All V. cholerae, V. parahaemolyticus and V. fluvialis strains possessed outer membrane protein W (ompW), thermostable direct haemolysin (tdh) and toxin regulatory protein (toxR) gene respectively. Hemolytic activity of V. cholerae exihibited different reaction isolated from seafood and plankton. The median lethal dose (LD50) of some V. cholerae strains was generally high. Conclusion: The result of the study suggested that the seafoods may act as an important reservoir of pathogenic vibrios and pose threat to human health.
Aims: Determine percentage positive by the Ziehl-Neelsen (ZN) stain; detect 245 base pair fragment of the IS6110 gene for Mycobacterium tuberculosis complex using INS1 and INS2 primers and 500 base pair fragment of the RvD1Rv2031c gene for M. bovis using the JB21 and JB22 primers by a multiplex PCR (M-PCR);compare number of positive samples detected by ZN stain and M–PCR; determine prevalence rates of Mycobacterium tuberculosis complex between the two animal species studied and estimate the rates of detecting agents of tuberculosis using Ziehl-Neelsen (ZN) technique and a Multiplex-Polymerase Chain Reaction (M-PCR) in lung samples of slaughtered cattle and goats in the study area. Place and Duration of Study: Samples were analyzed at the Central Diagnostic Laboratory and Molecular Biology Departments of the National Veterinary Research Institute Vom. This work was carried out between July-October 2010. Study Design: Experimental. Methodology: Our PCR amplified the 245 base pair (bp) fragment which is specific for this group of Mycobacterium while ZN exploited the acid-fast nature of the organisms. We examined one hundred lung samples, of cattle and goats, fifty for each. Results: Out of the lung samples from cattle, 15 (30%) were positive by ZN while 9 (18%) were positive by PCR. Among the ZN negative samples from cattle, one was positive by M-PCR. All the 50 samples from goats were negative by the two diagnostic tools used in this study. Results obtained in this study showed 0% TB infection in goats and 18% in cattle. Conclusion: The study showed a high infection rate of tuberculosis among cattle sampled in the area of study, as such, preventive and curative measures have to be stepped up in controlling the zoonosis.
Aim: To correlate H2O2 production of Lactobacillus species with the Nugent scores of young Nigerian women in order to assess their vaginal health. Study Design: Cross-sectional study. Place and Duration of Study: Departments of Medical Microbiology & Parasitology, Biochemistry and Obstetrics and Gynaecology, College of Medicine of the University of Lagos, between May and august 2009. Methodology: Ninety- seven isolates of Lactobacillus from eighty-two women without Bacterial vaginosis (BV) and fifteen women with BV were used for the study. BV was diagnosed using Nugent scoring method. Lactobacilli were isolated using MRS agar and categorized into facultative anaerobes and strict anaerobes. Hydrogen peroxide was detected and measured by titration using dilute sulphuric acid and reaction stopped with potassium permanganate. Results: Out of 97 isolates studied, 76 (78%) were facultative anaerobes, while 21 (22%) were strict anaerobes. The facultative anaerobes were obtained from 11 of 15 women with BV and 65 of 82 women without BV. Forty- nine (50.51%) of the 97 isolates produced H2O2. Forty- four of the H2O2 producers were from women without BV while five were from women with BV. Majority (67%) of the strains obtained from women with BV were non-hydrogen peroxide producing. Proportion of H2O2 producing Lactobacillus by Nugent score were 70%, 43% and 33% in negative, intermediate and BV Nugent scores respectively. There was no significant difference between the mean concentrations of H2O2 production in the various Nugent scores. Conclusion: The overall rate of hydrogen peroxide production was low. While the rates of hydrogen peroxide production correlated with Nugent scores, being highest in negative Nugent scores and lowest with BV scores, the concentration of hydrogen peroxide produced had no association with Nugent scores. The Nigerian women studied might have a relatively high susceptibility rate to vaginal infections.
Aims: The emergence of pathogenic bacterial strains with resistance to commonly used antibiotics has necessitated a search for novel types of antibacterial agents. The main objective of this study was to evaluate the synergistic action between honeys and the essential oils (EOs) against Pseudomonas aeruginosa (ATCC 27853). Study Design and Methodology: In the first step, the minimum inhibitory concentration (MIC) of each honeys and Eos were determined. In the second step, lower concentrations of honey than the MIC were mixed with a set of sub-MIC of EOs and then added to media to determine the minimum synergistic inhibitory concentration (MSIC). Place of Study: Laboratory Research onof Local Animal Products, Ibn-Khaldoun University, Tiaret, Algeria. Results: The results indicated that the essential oils and all varieties of honey were effective against P. aeruginosa. The effectiveness was correlated to the botanical origin of honey and EOs. Wild carrot honey and Origanum vulgaris EOs were the most effective against the tested bacteria with a MIC of 8% and 2% respectively. Adding EOs to honey decreases the MIC values and the isobolographic representation shows a synergistic action between the EOs and all varieties of honey. Conclusion: The current prevalence of antibacterial resistant species has led to a re-evaluation of the therapeutic use of ancient remedies, including honey and EOs, which may receive renewed recognition as wound and burn healers.
Aim:Iru is a popular West Africa fermented soup condiment which is also consumed without cooking as snack. This product is mainly fermented by Bacillus species. The hypolipidemic activities of Bacillus spp. isolated from iru have not been documented hence the aim of this study. Place and Duration of Study:Iru sample was bought in an open market in Iworoko-Ekiti, Nigeria and transferred to the Laboratory of the Department of Microbiology, Ekiti State University, Nigeria where other studies were carried out. The study was conducted between January and June, 2012. Methodology: The properties and invivo hypolipidemic potential of Bacillus species from iru were investigated using standard microbiological and haematological methods. Results: The cell free extracts of the Bacillus spp. did not produce significant inhibition on the selected Gram positive and Gram negative pathogens. Qualitative enzyme screening of the isolates showed all were haemolysin negative. Only B. subtilis was positive to gelatinase while all the isolates produced catalase and lipase. The average weight of the animals after inducement of hyper-cholesterolemia ranged between 60.5g - 95.3g. The amount of serum total cholesterol (TC) in the animals ranged between 124.9 mg/dl – 127.4 mg/dl while that of serum triglycerides (TG), high density protein (HDL) and low density protein (LDL) were 122.5 – 155.3 mg/dl, 10.0 – 15.3 mg/dl and 76.6 – 81.0 mg/dl respectively. The weights of hyper-cholesterolemia induced rats challenged with different species of Bacillus were relatively lower than those in the control group and also differ significantly from the control, at pË‚ 0.05. The values of TC, TG, and LDL were highest in the control (saline) group while the values in the treatment group ranged between 121.3 ± 1.5 and 102.3 ± 6.8 mg/dl for TC. The treatment groups recorded lower values of values for TG (104.7 ± 1.6 - 117.4 ± 9.1 mg/dl) and LDL (42.6 ±7.4 - 59.0 ± 10.2 mg/dl) compared to the control. B. subtilis had the highest values of TC but least amount of LDL. TG in all the groups was higher than TC, HDL and LDL. The TC/HDL and the LDL/HDL of the animals in the iru group was higher than the other treatment groups but lower than the control. Conclusion: Compared to the control, hypolipidemic activities of B. lichenliformis was the best followed by B. subtilis. Iru had the least hypo-cholesterolemic effect.
Background:Candida species are now recognized as major causative agents of hospital-acquired infection.
Aims: To evaluate the species distribution, biofilm formation,and antifungal susceptibility (amphotericin B, ketoconazole, and fluconazole) of Candida isolates. Place and Duration of Study: This is a Six-months Cross sectional study conducted in Al-ansar hospital, Al-Madinah, Saudi Arabia. Methodology: One hundred and three isolates of Candida spp. were cultured on Sabouraud dextrose agar (SDA). Candida spp. were identified by four standard methods, CHROMagar candida, cornmeal agar, germ tube test and API 20C. Detection of Biofilm formation was done by microtitre plate and antifungal susceptibility testing was done by disc diffusion. Results: C. albicans was the most common species 61%, followed by C. tropicalis 25%, C. lusitanaie 5%, C. parapsilosis 4%, C. glabrata 4%, and C. famata 1%. Biofilm formation was found to occur most frequently among non-albicans spp.(70%) than C. albicans (46%). All isolates were sensitive to amphotericin B and ketoconazole. Resistance to fluconazole was found in 22.5% of non-albicans spp. and 5% of C. albicans isolates. Conclusion: The present study proved that C. albicans is still the major isolate from urinary, vaginal and respiratory samples but non-albicans spp. predominate in the blood samples and from plastic devices. The non-albicans spp. were more biofilm - producers compared to C. albicans and C. tropicalis showed the highest score of biofilm intensity (grade 4+). The species isolated are less susceptible to fluconazole.
Aims: The present work aims to evaluate the antibacterial activity of essential oils of two different parts of Pistacia lentiscus (leaves and twigs) and to determine their chemical composition. Study Design: An experimental study Place and Duration of Study: 1- Laboratory of Microbial Biotechnology, Faculty of Science and Technology Saïss, Sidi Mohamed Ben Abdellah University, Fez, MOROCCO. 2- Laboratory of Aromatic Plants, Medicinal and Natural Substances, National Institute of Medicinal and Aromatic Plants, Sidi Mohamed Ben Abdellah University, Fez. MOROCCO. Between November 2012 and April 2013. Methodology: The study of antibacterial activity was performed on gram-negative bacteria Salmonella sp., Pseudomonas aeruginosa, Mycobacterium smegmatis and MycobacteriumMC² 155 aurum A+ and gram positive bacteria Staphylococcus aureus, Bacillus sp. and Enterococcus faecalis by the method of agar diffusion. The chemical composition of essential oils was identified by gas chromathography. Results: These oils presented an important antibacterial activity against Salmonella sp., Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus sp., Enterococcus faecalis, Mycobacterium aurum, Mycobacterium smegmatis. The use of gas chromatography enabled us to identify a total of 43 volatile components in the essential oil of twigs and 36 in leaves, representing respectively 95.9% and 86.8% of the chemical composition of these essences. These compositions are quantitatively and qualitatively different, which seems to be due to large structure differences of the two plant parts tested. Conclusion: Current study supports the traditional use of aromatic plants as antibacterial agents.
Aim: The study evaluated the inhibitory effect of fermentation products of β-mannanase-producing bacteria on selected poultry borne pathogens. Study Design: The first experiment, bacterial isolates previously confirmed positive for mannanase by plate assay technique were further screened for mannanase production in submerged state fermentation. In the second experiment, inhibitory effect of fermentation products of mannanase-producing bacteria on selected poultry pathogens was evaluated. Place and Duration of Study: Microbiology Research Laboratory, Federal University of Technology, Akure Nigeria between September 2011 and March 2012. Methodology: Bacterial isolates from agricultural wastes previously confirmed positive for mannanase activity by plate assay were further screened for their potential performance under submerged state fermentation and enzyme activity determined by dinitrosalicylic acid method. The inhibitory action of β-mannanase-producing bacteria was determined by supplementation of supernatant and plating method. Results: Isolate 1A showed highest mannanase activity (13.430 U/ml), displayed broad inhibition to selected poultry borne pathogens; Klebsiella oxytoca, Shigella alkalescens, Escherichia coli, Salmonella typhii, Staphylococcus aureus and Streptococcus sp. Apart from isolate 1A, fermentation products of other isolates generated from the mannolytic action of β-mannanase on mannan containing substrate displayed different percentage inhibition on selected poultry borne pathogens. Conclusion: The results suggested that fermentation products from β-mannanase-producing bacteria might possess antibacterial properties which could be applied in poultry farms.
Aims: The study was conducted to compare the efficacy of natural growth promoter AV/AGP/10 with antibiotic supplements on overall growth performance and intestinal micrometry of broiler birds. Study Design: Total of 150 healthy day old Vencob broiler chicks of nearly similar live body weight were equally divided into 5 groups of 30 birds each with three replicates in each group. All the groups were fed with basal diet. Group-I was positive control without any supplement, Group- II was supplemented with AV/AGP/10 @ 250g/ton of feed, Group-III supplemented with AV/AGP/10@500g/ton of feed, Group-IV supplemented with Bacitracin Methylene Dicyticylate @100g/ton of feed and Group-V supplemented with Oregostim @ 250g/ton of feed. Place and Duration of Study: the study was conducted in the department of Animal Nutrition, College of Veterinary and Animal sciences, Udgir, Dist. Latur, Maharashtra, India during the month of April- June 2012 for 42 days. The mean maximum daily temperature recorded at the time of trial was 41±2ºC and relative humidity (RH) 80.57 ± 1.50 %.
Methodology: the efficacy of the products was assessed on the basis of feed consumption, body weight gain, feed conversion ratio (FCR), metabolic trial / nutrient retention trial, intestinal micrometry and carcass yield / dressing percentage. Results: at the end of sixth week, significantly higher live body weight (1874.19, 1921.51, 1720.39 and 1673.58) with more economical FCR (1.74, 1.71, 1.78 and 1.78) along with marked improvement in digestibility of nutrients from supplementation of herbal growth promoter with equal competence as that of synthetic antibiotic was observed. The intestinal micrometry at day 21 and 42 also revealed better results with natural growth promoter as compared to synthetic growth promoter and control group in terms of villous height, width and crypts depth. Conclusion: Considering the overall trial results and harmful effects of antibiotic growth promoter such as bacterial resistance or undesired residues in animal products, the natural product AV/AGP/10 is better option as growth promoter and performance enhancers in broiler birds.
Aim: The study evaluated potential performance of different fungal isolates from agricultural by-products for mannanase production. Study Design: The first experiment, fungal isolates were screened for mannanase production on agar medium containing Locust Bean Gum (LBG) and total fungal count was conducted. In the second experiment, the fungal isolates were further screened for mannanase production in submerged state fermentation. Place and Duration of Study: Microbiology Research Laboratory Federal University of Technology, Akure and Postgraduate Research Laboratory, Obafemi Awolowo University Ile-Ife, Nigeria between September 2011 and March 2012. Methodology: The fungal isolates associated with some agricultural wastes were isolated on LBG containing agar medium by plate assay techniques and counted by standard microbiological methods. Mannanase production was conducted in submerged state fermentation (shaken & static) into which copra meal had been supplemented as the sole carbon source and enzyme activity was determined by dinitrosalicylic acid method. Results: In this study, 11 fungal isolates showed positive results with clear zone around their cultures. Fungal isolate 5A showed the highest activity ratio of 1.8, while the least was observed in isolate 9A12 with activity ratio of 0.64. The highest fungal counts were recorded in fermented coconut with 7.4×102 sfu/g, while cocoa pod and groundnut shell had no fungal growth. In terms of percentage occurrence of fungal isolates from selected agro-wastes, it was revealed that Rhizopus japonicus had the highest occurrence of 66.67%, while the same value of 8.33% was observed for Aspergillus fumigatus, A. glaucus, R. stolonifer and Trichosporonoides oedocephalis. In fermentation broth, all the 11 isolates displayed mannanase activity ranging from 0.370 to 21.667 U/ml for static and 0.278 to 3.982 U/ml for shaken condition, with the highest mannanase activity observed with isolate 5A for both culture conditions. According to the cultural characters and microscopic morphology, the isolate 5A being the highest mannanase producer was identified as the Aspergillus fumigatus. Conclusion: In this study, fungal isolates screened and evaluated for mannanase production from agricultural by-products elaborated considerable mannanase activity and this could be exploited for prebiotic preparation.
Aim: To investigate effect of microwave (MW) radiation on bacterial growth, enzyme activity (amylase and pectinase), and exopolysaccharide production. Study Design: The study was designed to investigate effect of MW radiation on bacterial growth, enzyme activity, and exopolysaccharide production. Particularly the non-thermal effects were focused. Thermal effects were avoided (minimized) by keeping the bacterial suspension in ice while exposing to MW radiation. Place and Duration of Study: Institute of Science, Nirma University, Ahmedabad, India, between November 2012 and May 2013. Methodology: The present study investigated the effect of MW (90 W) radiation on bacterial growth, enzyme activity (amylase and pectinase), and exopolysaccharide (EPS) production. Test parameters viz. growth, enzyme activity, and EPS production of populations originated from MW treated cells were compared to those originated from untreated control. Thermal effects of MW radiation were avoided (minimized) by placing inoculum vial(s) in a ice containing beaker during MW exposure. Results: MW treatment was found to be capable of altering bacterial growth, enzyme activity, and EPS production significantly. Amylase activity in B. subtilis suffered a heavy loss of 67.43% (P<0.01) following 6 min MW exposure. Pectinase activity in MW treated (4 min duration) B. subtilis was 169.92 times higher (P<0.01) than that of control. MW treatment for 4 min and 6 min duration were able to induce EPS production in Xanthomonas campestris by 46.15% (P<0.01) and 53.84% (P<0.05) respectively. Conclusion: MW treatment was found to alter growth, enzyme activity, and EPS production significantly in the test bacteria. This study positively suggests existence of non-thermal effects of MW radiation on biological entities. Further investigation on mode of action of these MW specific athermal effects, and on their genetic stability are warranted.
Aim: The work focused on the isolation and screening of mannanase-producing bacteria associated with selected agricultural wastes. Study Design: The first experiment, mannanase-producing bacteria were screened for mannanase production on Locust Bean Gum (LBG) agar medium and total bacterial count was determined. In the second experiment, the isolated bacteria were further screened for mannanase production in submerged state fermentation. Place and Duration of Study: Microbiology Research Laboratory Federal University of Technology, Akure and Postgraduate Research Laboratory, Obafemi Awolowo University Ile-Ife, Nigeria between September 2011 and March 2012. Methodology: The associated bacterial isolates were isolated on agar medium containing LBG and counted by standard microbiological methods. Quantitatively, mannanase production was conducted in mineral salt medium into which copra meal had been incorporated as the sole carbon source and enzyme activity was determined by dinitrosalicylic acid method. Results: The highest bacteria counts were recorded in compost from wood dust with 5.5×1011 cfu/g, while cassava peels had the least of 1.02×106 cfu/g. In this study, 23 bacterial isolates showed positive results with clear zone around the cultures. Bacterial isolate 1A showed the highest ratio of clear zone to colony, while the lowest was observed in isolate 4B. In liquid broth, all the 23 isolates displayed mannanase activity between 0.28 to13.89 U/ml for static and 0.56 to13.43 U/ml for shaken condition, with the highest mannanase activity observed with isolate IA for both culture conditions. In the comparative study between static and shaken conditions, it was revealed that shaken cultures exhibited better yield than static cultures. According to the morphological and biochemical studies, the isolate 1A was primarily identified as the Klebsiella edwardsii. Conclusion: In this investigation, bacterial isolates evaluated for mannanase production from agricultural wastes elaborated considerable mannanase activity and this could be applied in feed and prebiotic.
Seasonal population dynamics of bacteria, actinomycetes and fungi was studied in rhizosphere and non-rhizosphere soil of chir pine (Pinus roxburghii Sarg.) seedlings growing in polybags and in situ (forest). In greenhouse experiment bacteria and actinomycetes were present in higher numbers and their populations fluctuated with season. Fungal population, although lower in numbers, remained stable throughout the year. Population fluctuations with lower numbers were more prominent in rhizosphere and non-rhizosphere soils of forest plants. Differential bacterial population characteristics viz. sporeformers, fluorescent colony producers, methylene blue reducers, ammonifiers and glucose fermenters were also taken into account. The population of sporeformers was comparable with methylene blue reducers, which was higher than fluorescent colony producers, ammonifiers and glucose fermenters, respectively. The rhizosphere soil bacterial count of nursery seedlings ranged from 4.36 x 106 – 6.37 x 106 g-1 dry soil weight and from 8.8 x 105 – 2.64 x 106 g-1 soil on dry weight basis in forest plants during various seasons. Sporeformers were a magnitude lower than total bacterial population and fluorescent colony producers were magnitude lower than sporeformers. Actinomycetes count ranged from 6.0 x 105 – 3.02 x 106 g-1 dry soil weight in the nursery plants and from 6.4 x 105 – 1.17 x 106 g-1 dry soil weight in forest plants. Fungal population was a magnitude lower than bacterial and actinomycetes population, which ranged from 9.0 x 104 – 2.9 x 105 g-1 dry soil weight in the nursery plants and from 9.0 x 104 – 3.1 x 105 g-1 soil in forest plants. A similar trend of microbial population fluctuation but with lower numbers was observed in forest non-rhizosphere soil. Aims: To compare the seasonal population fluctuations of rhizosphere and non-rhizosphere soil for better understanding of population dynamics of soil microorganisms. Study Design: The observations were taken from nursery grown and forest grown seedlings. The microbial populations of pine seedling rhizosphere and non-rhizosphere soil were seasonally enumerated for one year at the intervals of three months for four times. Place and Duration of Study: The study was conducted at Sardar Bhagwan Singh (PG) Institute of Biomedical Sciences and Research (S.B.S.P.G.I.), Dehradun, Uttarakhand, India, situated at foothills of central Himalayas, for one year during January to December, 2005. Methodology: The seeds of Pinus roxburghii Sarg. were collected from natural chir pine forest from one healthy plant to minimize genetic variability in the experiment. The seeds were germinated on water agar medium and saplings were planted in polybags and kept in greenhouse nursery. Microbial colony forming units (CFUs) of the rhizosphere and non-rhizosphere soil of nursery grown and forest seedlings were enumerated for one year. Their populations were correlated with the meteorological data of the Dehradun valley. Results: The total bacterial population in terms of CFUs was comparatively higher in all seasons followed by actinomycetes, both these populations fluctuated with season. Fungal population, although lower in numbers, was consistent throughout the period. Microbial populations were found to be dependent on environmental factors like soil and air temperature, relative humidity and precipitation. The population of each microbial type reached maximum during third trimester, just after the end of monsoon season. Conclusion: The microbial population of chir pine rhizosphere soil and non-rhizosphere soil fluctuates seasonally. Microbial populations were found to be dependent on soil temperature, air temperature, precipitation and relative humidity.
Aims: To determine resistance rates and patterns of certain uropathogens, including E. coli, Klebsiella spp. and Pseudomonas spp., isolated from hospitalized urinary tract infections patients, to aminoglycoside antibiotics and to detect the most prevalent plasmid-mediated aminoglycoside modifying enzymes (AMEs). Methods: Uropathogenic isolates (150) were recovered from urine specimens of hospitalized UTI patients in Cairo, Egypt and identified by conventional methods. The recovered uropathogens (E. coli, Klebsiella spp. and Pseudomonas spp.) were tested for their susceptibility to gentamicin, tobramycin, amikacin, neomycin, netilmicin, and kanamycin by disc diffusion method. Plasmid-mediated aminoglycoside resistance was determined by transformation experiments as well as by using plasmids as templates for PCR screening of the AMEs-coding genes aph(3')-I, aac(6')-I, aac(3)-I, aac(3)-II and ant(2'')-I in all resistant isolates. Results: Of a total of 150 uropathogenic clinical isolates, 110 isolates were of the above mentioned genera and were selected for the current study. Sixty three isolates (57.2%) were resistant to at least one aminoglycoside antibiotic. Highest and lowest resistance rates were observed to kanamycin (53.6%) and amikacin (7.2%), respectively. The resistance rates to gentamicin, neomycin, tobramycin and netilmicin were 33.6%, 24.5%, 23.6% and 14.5%, respectively. AMEs-coding genes were detected on the plasmids of 93.6% of resistant isolates with prevalence rates of 53.9% for ant(2'')-I, 38% for both aac(6')-I and aac(3)-II and 33.3% for aph(3')-I, while aac(3)-I gene was not detected in any of the tested resistant isolates. Double and triple combinations of AMEs-coding genes were detected in ich49.2% of resistant isolates. Conclusion: A high prevalence of plasmid-mediated resistance to aminoglycoside antibiotics in Gram negative uropathogens from hospitalized patients was observed. Uropathogens may represent potential reservoirs of panaminoglycoside resistance in hospitals, having on their plasmids combinations of AMEs-coding genes. Good infection control measures in Egyptian hospitals together with periodic screening of prevalence rates of different resistance genes are required.
Aims: Human papillomavirus type 16 (HPV16) is the primary etiological agent of cervical cancer. The variations in the amino acid sequence of the HPV16 E6 and E7 oncoproteins are known to correlate with both their oncogenic potential and geographic distribution. Study Design: The present study was designed to analyze sequence variations in E6 and E7 genes of HPV16 in order to evaluate the intratype variants circulating in our population. Methodology: The entire E6 and E7 genes of 31 HPV16 isolates from Moroccan patients with cervical cancer were sequenced and analyzed. Results: Sequence analysis of HPV16-E6 showed a high prevalence (64.5%) of the African lineage. The European and the North-American variants were detected in respectively 19.4% and 16% of the HPV16 positive specimens. At the amino acid level, the most prevalent missense mutations revealed in the E6 gene were H78Y, Q14D, L83V, R10I and Q14H. Our data also showed that E7 appeared to be better conserved as compared to E6, with a high frequency of two silent variations at G789A and T795C nucleotides and one hot spot of E7 nucleotide variation A647G leading to N29S. Conclusion: The present study provides a new data on the genetic diversity of HPV16 and highlights the possible association between the high prevalence of HPV16 African variants and the high incidence of cervical cancer in Morocco.
Aims: To understand the mechanisms of Early Growth Response Protein 1 (Egr-1) induction upon HSV-1 lytic infection and its roles in regulating viral gene expression and replication. Study Design: Rabbit corneal cell line SIRC and other cell lines were infected by HSV-1 to investigate the Egr-1 induction and its occupancy on the viral genome in different conditions. UV-inactivated HSV-1 and a recombinant virus over-expressing Egr-1 were generated to evaluate the regulatory effects on viral gene expression and replication during the infection. Methodology: Egr-1 induction triggered by viral infection was determined by Western Blot analyses and immune-fluorescent microscopy. Real-time RT-PCR and a novel Cignal™ Reporter Assay were used for quantitative measurement of Egr-1 expression. Chromatin Immuno-precipitation (ChIP) was performed to address the Egr-1 occupancy to the viral regulatory sequences and the influence on viral replication was assessed by plaque assays. Results: Our results indicated that Egr-1 expression requires viral gene expression since the UV-inactivated HSV-1 failed to produce Egr-1 protein. Blockade of viral replication did not block the Egr-1 protein synthesis, supporting the hypothesis that HSV-1 replication was not essential for Egr-1 production. Chromatin immune-precipitation (ChIP) and RT-PCR assays demonstrated that induced Egr-1 was able to interact with key regulatory elements near HSV-1 immediate-early (IE) genes and promote viral gene expression. Recombinant virus overexpressing Egr-1 revealed that Egr-1 enhanced the viral replication and the release of infectious virus. Conclusion: Together these results concluded that HSV-1 triggers the expression of an important host transcription factor Egr-1 via a unique mechanism and benefit the viral gene expression and replication.
Aims: Selective enumeration, isolation and characterization of lactic acid bacteria from different raw milk sources with special elucidation to lactic acid production. Study Design: Serial dilution for strain isolation from raw milk samples to be done followed by complete morphological, biochemical and molecular characterization aiming to study the microbial behavior to ferment different sugar and producing lactic acid as end product. Place and Duration of Study: Microbiology Biotechnology Laboratory, Tropilite Foods Pvt. Ltd. Gwalior (M.P)-INDIA from Sep 2012 to April 2013. Methodology: Milk samples from cow, sheep, goat, camel and buffalo were collected from the surrounding area of Gwalior district of Madhya Pradesh, India, from local milk suppliers. 13 potent strains in terms of lactic acid production were selected for analysis. Gram Staining, Catalase activity, Sugar fermentation, growth at high (45ºC) and low (10ºC) temperatures along with growth in different NaCl concentrations was observed with all isolates. Significant molecular characterization was done to determine the homology between different isolates of lactic acid bacteria. Results and Conclusion: Five potent lactic acid bacteria strains Streptococcus thermophilus from goat milk, Lactococcus lactis from buffalo milk, Streptococcus gallolyticus from camel milk, Streptococcus thermophilus from cow milk, Lactobacillus delbrueckii from sheep milk were identified capable of producing lactic acid in generous amount. Also all isolated strains from goat milk were found efficient in terms of lactic acid production when compared to other raw milk sources.
The continued decline in soil fertility, high fertilizer costs and the need to implement environmental friendly agricultural systems are some of the world’s major strategic concerns. Soil microorganisms are part of the soil ecosystem and are reported to contribute in soil fertility improvement. This paper is aimed at highlighting their contributions in alleviating soil fertility decline. The Rhizobium/legume symbiosis, a well known association contributes substantial amounts of biologically fixed nitrogen to cropping systems and significant benefits on yields of crops that follow in rotation. Soil microorganisms such as bacteria and fungi contribute to plant phosphorus nutrition through solubilization of sparingly soluble Al, Fe and Ca phosphates, and mineralization of phosphorus from organic substances. Solubilization is mainly achieved through production of organic acids, chelation and ligand exchange, and other pH lowering mechanisms whereas mineralization is achieved through production of enzymes such as phytases and phosphatases. Mycorrhizal associations are reported to contribute to plant phosphorus nutrition through increasing root surface area for soil exploration, production of phosphorus solubilizing enzymes and acids. Mycorrhizal fungi and bacteria also solubilize other nutrients such zinc, copper, potassium and calcium from their precipitated or sparingly soluble forms. Microorganisms also contribute to soil fertility improvement through their roles in composting. They are currently isolated, studied and packaged as biofertilizers and used to supplement chemical fertilizers. It can be noted that thorough exploitation of microbial activities can contribute to balanced plant nutrition. However, poor soil management practices limit realization of potential benefits from soil microorganisms whereas biofertilizer technology development in developing countries such as Sub-Saharan African countries is derailed because of lack of awareness, infrastructure and human capacity. From this study it can be noted that intensifying soil management practices that maximize microbial activities can go a long way in improving soil fertility with minimal use of chemical fertilizers. On the other hand there is a need to improve both human and infrastructure capacity in poor countries such as those in Sub-Saharan Africa so as to manage research in biofertilizer technologies. Awareness and dissemination of information on the importance of biofertilizers, intensifying effective microbial inoculation where it deemed to give good response and systematic evaluation of economic viability of biofertilizer technologies are other areas that need to be addressed.