Search results for: halotolerant pathogenic bacteria

Authors: Kumudini Apsara Munasinghe, Devin Gregory Lissau, Ryan Thomas Wade

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Fresh fruits and vegetables are rich in vitamins, minerals, and fibers and help maintain a healthy weight over high-calorie food. Eating fruits and vegetables protects us from free radicals produced by metabolic reactions and safeguards us from cardiovascular disease and cancer. However, there has been an increased concern about foodborne diseases tied to contaminated farmers’ market produce. In addition, very little information is available about the contribution of eating raw fruits and vegetables to human exposure to antibiotic-resistant bacteria. This research aims to identify bacteria isolated from farmers’ market fruits and vegetables and understand their antibiotic resistance. Vegetables and fruits were collected from farmers’ markets around Frostburg and Cumberland areas in Maryland and transported to the microbiology lab at Frostburg State University for the isolation of bacteria. Bacteria were extracted from tomatoes, cucumber, strawberry, and lettuce using Tryptic soy broth overnight at 37°C, and Tryptic Soy agar was used for the streak plate technique to isolate bacteria. Pure cultures were used to identify bacteria using biochemical reactions after conducting Gram staining technique. The research used many biochemical reactions, including Mannitol Salt agar, MacConkey agar, and Eosin Methylene blue agar, for identification. Antibiotic sensitivity was tested for many different types of antibiotics, including amoxicillin, penicillin, tetracycline, ampicillin, and erythromycin. Most prevalent bacteria in the isolates were Staphylococcus, Bacillus, Micrococcus, Enterococcus, Enterobacter, Citrobacter, and other bacteria from the family Enterobacteriaceae. The data obtained from this research will be useful to educate and train farmers and individuals involved in post-harvest processes such as transportation and selling in farmers’ markets. Further results for bacterial antibiotic resistance will be obtained, and unculturable bacteria will be identified by next-generation DNA sequencing.

Keywords: antibiotic resistance, farmers markets, fruits, bacteria, vegetables

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Authors: Bassam Al Johny

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Halophilic bacteria are organisms which thrive in salt-rich environments, such as salt lakes, solar salterns and salt mines which contain large populations of these organisms. In biotechnology, such salt-tolerant bacteria are widely used for the production of valuable enzymes, and more than a thousand years ago humans began using salt to cure and thereby preserve perishable foods and other materials, such as hides; halophiles can be detrimental to the preservation of salt brine cured hides. The aim of this review is to provide an overview of the taxonomy of these organisms including novel isolates from rock salt, and also to discuss their current and future biotechnological and environmental uses.

Keywords: hypersaline environments, halophilic bacteria, environmental application, industrial application

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Authors: Hamoud Alshammari, Adnan Almezani, Hamdan Alshammari, Faris Alharbi

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In this study we have examined of antimicrobial activity for unripe Juglan Regia phenolic extracts against a wide range of pathogenic microorganisms. Walnut (Juglans regia L.) is a member of Juglandaceae family used as a remedy in folk medicine. Leaves, barks, fruits and husk (peel) reported to harbor distinctive medical effect. In our study, we examined the anti-microbial effect against a set of gram positive and negative bacteria and even we have tested them against eukaryotic candida strains in a concentration gradual manner. Ethyl acetate extract of J. regia had the best antibacterial activity when compared with ciprofloxacin. The Minimum inhibition concentration for S. aureus, P. aerogenosa and S. epidermidis MIC was 0.85 mg/mL.

Keywords: antimicrobial, J. regia, S. aureus, phytochemistry

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Authors: Ayoub Azizi-Shotorkhoft, Tahereh Mohammadabadi, Hosein Motamedi, Morteza Chaji, Hasan Fazaeli

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This study was conducted to investigate nutritive value of wheat straw processed with termite gut symbiotic bacteria with lignocellulosic-degrading potential including Bacillus licheniformis, Ochrobactrum intermedium and Microbacterium paludicola in vitro. These bacteria were isolated by culturing termite guts contents in different culture media containing different lignin and lignocellulosic materials that had been prepared from water-extracted sawdust and wheat straw. Results showed that incubating wheat straw with all of three isolated bacteria increased (P<0.05) acid-precipitable polymeric lignin (APPL) compared to control, and highest amount of APPL observed following treatment with B. licheniformis. Highest and lowest (P<0.05) in vitro gas production and ruminal organic matter digestibility were obtained when treating wheat straw with B. licheniformis and control, respectively. However, other fermentation parameters such as b (i.e., gas production from the insoluble fermentable fractions at 144h), c (i.e., rate of gas production during incubation), ruminal dry matter digestibility, metabolizable energy, partitioning factor, pH and ammonia nitrogen concentration were similar between experimental treatments (P>0.05). It is concluded that processing wheat straw with isolated bacteria improved its nutritive value as ruminants feed.

Keywords: termite gut bacteria, wheat straw, nutritive value, ruminant

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Authors: B. S. Chandrashekar, M. K. Prasannakumar, P. Buela Parivallal, Sahana N. Banakar, Swathi S. Patil, H. B. Mahesh, D. Pramesh

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Bacterial soft rot is one of the most often seen diseases in many plant species globally, resulting in considerable yield loss. Radish roots with dark water-soaked lesions, maceration of tissue, and a foul odour were collected in the Kolar region, India. Two isolates were obtained from rotted samples that demonstrated morphologically unpigmented, white mucoid convex colonies on nutrient agar medium. The isolated bacteria (RDH1 and RDH3) were gram-negative, rod-shaped bacteria with biochemically distinct characteristics similar to the type culture of Enterobacter cloacae ATCC13047 and Bergy's handbook of determinative bacteriology. The 16s rRNA gene was used to identify Enterobacter species. On carrot, potato, tomato, chilli, bell pepper, knolkhol, cauliflower, cabbage, and cucumber slices, the Koch′s postulates were fulfilled, and the pathogen was also pathogenic on radish, cauliflower, and cabbage seedlings were grown in a glasshouse. After 36 hours, both isolates exhibited a hypersensitive sensitivity to Nicotianatabacum. Semi-quantitative analysis revealed that cell wall degrading enzymes (CWDEs) such as pectin lyase, polygalacturonase, and cellulase (p=1.4e09) contributed to pathogenicity, whereas isolates produced biofilms (p=4.3e-11) that help in host adhesion. This is the first report in India of radish soft rot caused by E. cloacae.

Keywords: soft rot, enterobacter cloacae, 16S rRNA, nicotiana tabacum, and pathogenicity

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Authors: Ramutė Miseikienė, Saulius Tusas

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Somatic cell count and bacteria count are the main indicators of cow milk quality. The aim of this study was to analyze and compare parameters of milk quality in different-sized cows herds. Milk quality of ten dairy cows farms during one year period was analyzed. Dairy farms were divided into five groups according to number of cows in the farm (under 50 cows, 51–100 cows, 101–200 cows, 201–400 cows and more than 400 cows). The averages of somatic cells bacteria count in milk and milk freezing temperature were analyzed. Also, these parameters of milk quality were compared during outdoor (from May to September) and indoor (from October to April) periods. The largest number of SCC was established in the smallest farms, i.e., in farms under 50 cows and 51-100 cows (respectively 264±9,19 and 300±10,24 thousand/ml). Reliable link between the smallest and largest dairy farms and farms with 101-200 and 201-400 cows and count of somatic cells in milk has not been established (P > 0.05). Bacteria count had a low tendency to decrease when the number of cows in farms increased. The highest bacteria number was determined in the farms with 51-100 cows and the the lowest bacteria count was in milk when 201-400 and more than 401 cows were kept. With increasing the number of cows milk maximal freezing temperature decreases (significant negative trend), i. e, indicator is improving. It should be noted that in all farms milk freezing point never exceeded requirements (-0.515 °C). The highest difference between SCC in milk during the indoor and outdoor periods was established in farms with 201-400 cows (respectively 218.49 thousand/ml and 268.84 thousand/ml). However, the count of SC was significantly higher (P < 0.05) during outdoor period in large farms (201-400 and more cows). There was no significant difference between bacteria count in milk during both – outdoor and indoor – periods (P > 0.05).

Keywords: bacteria, cow, farm size, somatic cell count

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Authors: Muhammad Adeel Arshad, Shaukat Ali Bhatti, Faiz-ul Hassan

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Manipulating microbial ecosystem in the rumen is considered as an important strategy to optimize production efficiency in ruminants. In the past, antibiotics and synthetic chemical compounds have been used for the manipulation of rumen fermentation. However, since the non-therapeutic use of antibiotics has been banned, efforts are being focused to search out safe alternative products. In tropics, crop residues and forage grazing are major dietary sources for ruminants. Poor digestibility and utilization of these feedstuffs by animals is a limiting factor to exploit the full potential of ruminants in this area. Hence, there is a need to enhance the utilization of these available feeding resources. One of the potential strategies in this regard is the use of direct-fed microbes. Bacteria and fungi are mostly used as direct-fed microbes to improve animal health and productivity. Commonly used bacterial species include lactic acid-producing and utilizing bacteria (Lactobacillus, Streptococcus, Enterococcus, Bifidobacterium, and Bacillus) and fungal species of yeast are Saccharomyces and Aspergillus. Direct-fed microbes modulate microbial balance in the gastrointestinal tract through the competitive exclusion of pathogenic species and favoring beneficial microbes. Improvement in weight gain and feed efficiency has been observed as a result of feeding direct-fed bacteria. The use of fungi as a direct-fed microbe may prevent excessive production of lactate and harmful oxygen in the rumen leading to better feed digestibility. However, the mechanistic mode of action for bacterial or fungal direct-fed microbes has not been established yet. Various reports have confirmed an increase in dry matter intake, milk yield, and milk contents in response to the administration of direct-fed microbes. However, the application of a direct-fed microbe has shown variable responses mainly attributed to dosages and strains of microbes. Nonetheless, it is concluded that the inclusion of direct-fed microbes may mediate the rumen ecosystem to manage lactic acid production and utilization in both clinical and sub-acute rumen acidosis.

Keywords: microbes, roughages, rumen, feed efficiency, production, fermentation

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Authors: Arun Jyothi Bheemagani, Chakrapani Pullagummi, Anupalli Roja Rani

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Exploration of the chemical constituents of the plants and pharmacological screening may provide us the basis for the development of novel agents. Plants have provided us some of the very important life saving drugs used in the modern medicine. The aim of our work was to screen the phytochemical constituents and antimicrobial and antioxidant activities of methanol extract of leaves of Annona cherimola Mill plant from Tirumala forest, Tirupathi. It was originally called Chirimuya by the Inca people who lived where it was growing in the Andes of South America, is an edible fruit-bearing species of the genus Annona from the family Annonaceae. Annona cherimola Mill is a multipurpose tree with edible fruits and is one of the sources of the medicinal products. The antibacterial activity was measured by agar well diffusion method; the diameter of the zone of bacterial growth inhibition was measured after incubation of plates. The inhibitory effect was studied against the pathogenic bacteria (Klebsiella pneumonia, Bacillus subtilis, Staphylococcus aureus and Escherichia coli (E. coli). Antioxidant assays were also performed for the same extracts by spectrophotometric methods using known standard antioxidants as reference. The studied plant extracts were found to be very effective against the pathogenic microorganisms tested. The methanolic extract of Annona cherimola Mill from showed maximum activity against Escherichia coli and Staphylococcus aureus and the least concentration required showing the activity was 0.5mg/ml. Phytochemical screening of the plants revealed the presence of flavonoids, alkaloids, steroids and carbohydrates. Good presence of antioxidants was also found in the methanolic extracts.

Keywords: annona cherimola, phytochemicals, antioxidant and antibacterial activity, methanol extract

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Authors: W. Mohammed Saeed, A. J. Mcbain, S. M. Cruickshank, C. A. O’Neill

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In the gut, probiotics have been shown to protect epithelial cells from pathogenic bacteria through a number of mechanisms: 1-Increasing epithelial barrier function, 2-Modulation of the immune response especially innate immune response, 3-Inhibition of pathogen adherence and down regulation of virulence factors. Since probiotics have positive impacts on the gut, their potential effects on other body tissues, such as skin have begun to be investigated. The purpose of this project is to characterize the potential of probiotic bacteria lysate as therapeutic agent for preventing or reducing the S. aureus infection. Normal human primary keratinocytes (KCs) were exposed to S. aureus (106/ml) in the presence or absence of L. rhamnosus GG lysate (extracted from 108cfu/ml). The viability of the KCs was measured after 24 hours using a trypan blue exclusion assay. When KCs were treated with S aureus alone, only 25% of the KCs remained viable at 24 hours post infection. However, in the presence of L. rhamnosus GG lysate the viability of pathogen infected KCs increased to 58% (p=0.008, n=3). Furthermore, when KCs co-exposed, pre- exposed or post-exposed to L. rhamnosus GG lysate, the viability of the KCs increased to ≈60%, the L. rhamnosus GG lysate was afforded equal protection in different conditions. These data suggests that two possible separate mechanisms are involved in the protective effects of L. rhamnosus GG such as reducing S. aureus growth, or inhibiting of pathogenic adhesion. Interestingly, a lysate of L rhamnosus GG provided significant reduction in S. aureus growth and adhesion of S. aureus that being viable following 24 hours incubation with S aureus. Therefore, a series of Liquid Chromatography (RP-LC) methods were adopted to partially purify the lysate in combination with functional assays to elucidate in which fractions the efficacious molecules were contained. In addition, the Mass Spectrometry-based protein sequencing was used to identify putative proteins in the fractions. The data presented from purification process demonstrated that L. rhamnosus GG lysate has the potential to protect keratinocytes from the toxic effects of the skin pathogen, S. aureus. Three potential mechanisms were identified: inhibition of pathogen growth; competitive exclusion; and displacement of the pathogen from keratinocyte binding sites. In this study, ‘moonlight’ proteins were identified in the current study’s MS/MS data for L. rhamnosus GG lysate, which could elucidate the ability of lysate in the competitive exclusion and displacement of S. aureus from keratinocyte binding sites. Taken together, it can be speculated that L. rhamnosus GG lysate utilizes different mechanisms to protect keratinocytes from S. aureus toxicity. The present study indicates that the proteinaceous substances are involved in anti-adhesion activity. This is achieved by displacing the pathogen and preventing the severity of pathogen infection and the moonlight proteins might be involved in inhibiting the adhesion of pathogens.

Keywords: lysate, fractions, adhesion, L. rhamnosus GG, S. aureus toxicity

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Authors: Hossein Mostafavi

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A diversity of biological activities and pharmaceutical uses have been attributed to gallic acid derivatives such as antibacterial, anticancer, anti inflammatory. A series of gallic acid derivatives were synthesized, and their structure was confirmed by FT-IR, HNMR, CNMR, elemental analysis. In vitro biological activity of compounds was determined against Proteus vulgaris ATCC 7829, Escherichia coli ATCC 25922, as (Gram-negative) bacteria and bacillus cereus ATCC 11778, Staphylococus aureus ATCC 6538 as (Gram-positive) bacteria. Antibacterial susceptibility tests were done by use of the paper disc diffusion method on Mueller Hinton agar (Merck). Chloramiphenicol, Penicilline, Streptomycin and Tetracycline were standard reference antibiotics. The zone of inhibition against bacteria was measured after 24 hours at 37 °C. Compounds 3, 4, 5 were the main antibacterial compounds against Gram-negative bacteria but not Gram-positive.

Keywords: gallic acid derivatives, antibacterial, antibiotics, inhibition

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Authors: G. T. Ramos-Escobedo, E. T. Pecina-Treviño, L. F. Camacho-Ortegon, E. Orrantia-Borunda

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The mineral bioflotation represents a viable alternative for the evaluation of new processes benefit alternative. The adsorption bacteria on minerals surfaces will depend mainly on the type of the microorganism as well as of the studied mineral surface. In the current study, adhesion of S. carnosus on coal was studied. Several methods were used as: DRX, Fourier Transform Infra Red (FTIR) adhesion isotherms and kinetic. The main goal is the recovery of organic matter by the microflotation process on coal particles with biological reagent (S. carnosus). Adhesion tests revealed that adhesion took place after 8 h at pH 9. The results suggest that the adhesion of bacteria to solid substrates can be considered an abiotic physicochemical process that is consequently governed by bacterial surface properties such as their specific surface area, hydrophobicity and surface functionalities. The greatest coal fine flotability was 75%, after 5 min of flotation.

Keywords: fine coal, bacteria, adhesion, recovery organic matter

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Authors: Maryam Beiranvand, Sajad Yaghoubi

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Background: Some microbes can colonize plants’ inner tissues without causing obvious damage and can even produce useful bioactive substances. In the present study, the diversity of the endophytic bacteria associated with medicinal plants from Iran was investigated by culturing techniques, molecular gene identification, as well as measuring them for antibacterial activity. Results: In the spring season from 2013 to 2014, 35 herb pharmacology samples were collected, sterilized, meshed, and then cultured on selective media culture. A total of 199 endophytic bacteria were successfully isolated from 35 tissue cultures of medical plants, and sixty-seven out of 199 bacterial isolates were subjected to identification by the 16S rRNA gene sequence analysis method. Based on the sequence similarity gene and phylogenetic analyses, these isolates were grouped into five classes, fourteen orders, seventeen families, twenty-one genera, and forty strains. The most abundant group of endophytic bacteria was actinobacterial, consisting of thirty-two (47%) out of 67 bacterial isolates. Ten (22.3%) out of 67 bacterial isolates remained unidentified and classified at the genus level. The signature of the 16S rRNA gene formed a distinct line in a phylogenetic tree showing that they might be new species of bacteria. One (5.2%) out of 67 bacterial isolates was still not well categorized. Forty-two out of 67 strains were candidates for antimicrobial activity tests. Nineteen (45%) out of 42 strains showed antimicrobial activity multidrug resistance (MDR); thirteen (68%) out of 19 strains were allocated to classes actinobacteria. Four (21%) out of 19 strains belonged to the Bacillaceae family, one (5.2%) out of 19 strains was the Paenibacillaceae family, and one (5.2%) out of 19 strains belonged to the Pseudomonadaceae family. The other twenty-three strains did not show inhibitory activities. Conclusions: Our research showed a high-level phylogenetic diversity and the intoxicating antibiotic activity of endophytic bacteria in the herb pharmacology of Iran.

Keywords: Antibacterial activity, endophytic bacteria, multidrug-resistant bacteria, whole genom sequencing

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Authors: Mansur Abdulrasheed, Hussein I. Ibrahim, Ahmed F. Umar

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Nitrification, the microbial oxidation of ammonia to nitrate (NO3-) via nitrite (NO2-) is a vital process in the biogeochemical nitrogen cycle and is performed by two distinct functional groups; ammonia oxidisers (comprised of ammonia oxidising bacteria (AOB) and ammonia oxidising archaea (AOA)) and nitrite oxidising bacteria. Autotrophic nitrification is said to occur in acidic soils, even though most laboratory cultures of isolated ammonia and nitrite oxidising bacteria fail to grow below neutral pH. Published studies revealed that soil pH is a major driver for determining the distribution and abundance of AOB and AOA. To determine whether distinct populations of nitrite oxidising bacteria within the lineages of Nitrospira and Nitrobacter are adapted to a particular range of pH as observed in ammonia oxidising organisms, the community structure of Nitrospira-like and Nitrobacter-like NOB were examined across a pH gradient (4.5–7.5) by amplifying nitrite oxido-reductase (nxrA) and 16S rRNA genes followed by denaturing gradient gel electrophoresis (DGGE). The community structure of both Nitrospira and Nitrobacter changed with soil pH, with distinct populations observed in acidic and neutral soils. The abundance of Nitrospira-like 16S rRNA and Nitrobacter-like nxrA gene copies contrasted across the pH gradient. Nitrobacter-like nxrA gene abundance decreased with increasing soil pH, whereas Nitrospira-like 16S rRNA gene abundance increased with increasing pH. Findings indicated that abundance and distributions of soil NOB is influence by soil pH.

Keywords: nitrospira, nitrobacter, nitrite-oxidizing bacteria, nitrification, pH, soil

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Authors: V. Verma, Syed Riyaz-ul-Hassan

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Staphylococcus aureus is one of the most commonly found pathogenic bacteria and is hard to eliminate from the human environment. It is responsible for many nosocomial infections, besides being the main causative agent of food intoxication by virtue of its variety of enterotoxins. Routine detection of S. aureus in food is usually carried out by traditional methods based on morphological and biochemical characterization. These methods are time-consuming and tedious. In addition, misclassifications with automated susceptibility testing systems or commercially available latex agglutination kits have been reported by several workers. Consequently, there is a need for methods to specifically discriminate S. aureus from other staphylococci as quickly as possible. Data on protocols developed using molecular means like PCR technology will be presented for rapid and specific detection of this pathogen in food, clinical and environmental samples, especially milk.

Keywords: food Pathogens, PCR technology, rapid and specific detection, staphylococcus aureus

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Authors: Ahdieh Alijani, Mina Zarrabi, Abdollah Jamshidi, Jamshid Razmyar

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Avian intestinal spirochetosis (AIS) is caused by spiral-shaped Gram-negative Brachyspira spp. in poultry and is known as a cause of diarrhea, low egg production and increased the occurrence of dirty eggs in layer hens. In this study the presence of some Brachyspira spp. was investigated in laying hens. A total of 100 cloacal swab samples were individually collected from 20 laying hen flocks showing fecal egg staining in northeastern Iran. By culture and morphologic examination, 41 samples (41%) from 20 flocks were positive but by using genus–specific PCR only 37 (37%) samples were confirmed as Brachyspira spp. Using species-specific primers, single colonization was identified in 18 samples associated with B. pilosicoli (48.6%) while single colonization with B. intermedia was found in only two samples (5.4%). Simultaneous colonization by B. intermedia and B. murdochii was detected in 3 samples (8.1%). B. pilosicoli was the most prevalent species in concurrent colonization in 11 cases (29.7%). Finally, co-colonization by B. intermedia and B. innocens was identified in 3 samples (8.1%). The results of this study show the colonization of different species of Brachyspira with the dominance of B. pilosicoli in layer hens. In simultaneous colonization with pathogenic and non-pathogenic species the symptoms of intestinal spirochetosis were reduced, suggesting a competitive role in preventing and reducing the colonization of pathogenic species.

Keywords: intestinal spirochetosis, Brachyspira, laying hen, PCR

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Authors: Idrees Khan, Zhang Baoliang

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Kaolin clay (KC) supported Zinc oxide (ZnO/KC) and ZnO nanoparticles (NPs) were prepared by a chemical reduction process and used for the photodegradation of methyl red (MR) as photocatalysts. Due to the interlayered porous structure of KC, we achieved a perfect association between ZnO NPs and KC. SEM image showed the irregular morphology of ZnO NPs, while ZnO/KC NCs were predominately round-shaped. Moreover, in both cases, NPs were present in dispersed and agglomerated forms with an average particle size way below 100 nm. The results acquired from photodegradation analyses showed that ZnO NPs and ZnO/KC NCs degraded about 82% and 99% of MR under UV light in a short irradiation time within 10 min. The recovered and re-recovered ZnO NPs and ZnO/KC NCs were also considerably photodegraded MR in an aqueous medium. The same NPs also exhibit promising bioactivities against two pathogenic bacteria, i.e., Citrobacter and Providencia. ZnO/KC NCs' antioxidant activity reached a reasonable 70% compared to the 88% activity of the standard ascorbic acid.

Keywords: nanoparticles, photocatalyst, photodegradation, zinc oxide, methyl red

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Authors: L. Lamola, E. Manolas, A. Krause

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Background: The incidence of childhood cancer incidence is increasing gradually in low-middle income countries, such as South Africa. Globally, there is an extensive range of familial- and hereditary-cancer syndromes, where underlying germline variants increase the likelihood of developing cancer in childhood. Next-Generation Sequencing (NGS) technologies have been key in determining the occurrence and genetic contribution of germline variants to paediatric cancer development. We aimed to design and evaluate a candidate gene panel specific to inherited cancer-predisposing genes to provide a comprehensive insight into the contribution of germline variants to childhood cancer. Methods: 32 paediatric patients (aged 0-18 years) diagnosed with a malignant tumour were recruited, and biological samples were obtained. After quality control, DNA was sequenced using an ion Ampliseq 50 candidate gene panel design and Ion Torrent S5 technologies. Sequencing variants were called using Ion Torrent Suite software and were subsequently annotated using Ion Reporter and Ensembl's VEP. High priority variants were manually analysed using tools such as MutationTaster, SIFT-INDEL and VarSome. Putative identified candidates were validated via Sanger Sequencing. Results: The patients studied had a variety of cancers, the most common being nephroblastoma (13), followed by osteosarcoma (4) and astrocytoma (3). We identified 10 pathogenic / likely pathogenic variants in 10 patients, most of which were novel. Conclusions: According to the literature, we expected ~10% of our patient population to harbour pathogenic or likely pathogenic germline variants, however, we reported about 3 times (~30%) more than we expected. Majority of the identified variants are novel; this may be because this is the first study of its kind in an understudied South African population.

Keywords: Africa, genetics, germline-variants, paediatric-cancer

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Authors: Ganga Sagar Bhattarai, Swastika Gurung

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Antibiotics were being misused in both humans and animals, which led to the development of multidrug-resistant microorganisms. Antibiotic abuse is likely rampant among goats, cows, and buffaloes in order to boost growth and reduce production losses. The aim of this study is to know the multidrug resistance (MDR) bacteria in goats, cows, and buffaloes. Out of 68 samples that were examined, S. aureus, Bacillus spp., E. coli, Shigella spp., Klebsiella spp., S. epidremidis, and Salmonella spp. were isolated. S. aureus was the highest isolated bacteria (91.17%), Bacillus spp. (61.76%), E. coli (48.52%), Shigella spp. (22.05%), Klebsiella spp. (17.64%), S. epidermidis (13.23%), and the Salmonella spp. (7.35%). Salmonella spp. and E. coli showed multidrug resistance to at least four antibiotics, including Amoxicillin, Tetracycline, Piperacillin, and Ciprofloxacin, in Salmonella and to at least three antibiotics, including Amoxicillin, Tetracycline, and Nalidic acid. The highest resistance bacteria Salmonella spp. showed (57.14%) E. coli and Bacillus spp. showed (42.85%) S. aureus, S. epidermidis, and Shigella spp. showed (28.57%), and Klebsiella spp. showed (14.28%). This study showed that antibiotic-resistant bacteria with high levels of Amoxicillin, Penicillin, and Tetracycline resistance are present in healthy farm animals such as goats, cows, and buffaloes. Options for antibiotic therapy in both humans and animals will likely be limited as a result. The use, distribution, storage, and sale of antibiotics in veterinary practices must consequently be under strict control.

Keywords: multidrug resistance, multidrug resistance bacteria, susceptibility testing, bacterial infections

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Authors: Ali Amiri, Arash Esfandiari, Elham Zarenezhad

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We report here an efficient and rapid method for the preparation of 3,4-dihydro-1H-quinoxalin-2-ones and 1H‑quinolin-2-ones that involves grinding of o-, m-, or p‑phenylenediamine and three dialkyl acetylenedicarboxylates using a pestle and mortar. This solvent-free approach requires only a few minutes of reaction time. This type of reaction is expected to be the most economical method since neither catalyst nor solvent is used. Finally, all synthesised compounds were screened for antimicrobial activity against two Gram-positive bacteria (Pseudomonas aeruginosa PTCC 1077, Escherichia coli PTCC1330) and two Gram-negative bacteria (Staphylococcus aureus PTCC 1133, Bacillus cereus PTCC 1015) and their activity. Compared with gentamycin and ampicillin as reference drugs for Gram-negative and Gram-positive bacteria, respectively. The minimum inhibitory concentration (MIC) of the synthesised compounds and reference drugs were determined by the microdilution method. Good antibacterial activity was observed for 3,4-dihydro-1H-quinoxalin-2-ones against all species of Gram-positive and Gram-negative bacteria, and1H‑quinolin-2-ones showed good antibacterial activity against two Gram-positive bacteria.

Keywords: quinolin, quinoxalin, anti-bacterial activity, minimum inhibitory concentration (MIC)

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Authors: Prachi Choudhary, Jai Gopal Sharma

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Microbial cultures determine the type of organism, its abundance in the tested sample, or both. It is one of the primary diagnostic methods of microbiology. It is used to determine the cause of infectious disease by letting the agent multiply in a predetermined medium. Different bacteria produce colonies that may be very distinct from the bacterial species that produced them. To culture any pathogen or microorganism, we should first know about the types of media used in microbiology for culturing. Sometimes sub culturing is also done in various microorganisms if some mixed growth is seen in culture. Nearly 3 types of culture media based on consistency – solid, semi-solid, and liquid (broth) media; are further explained in the report. Then, The Five I's approach is a method for locating, growing, observing, and characterizing microorganisms, including inoculation and incubation. Isolation, inspection, and identification. For identification of bacteria, we have to culture the sample like urine, sputum, blood, etc., on suitable media; there are different methods of culturing the bacteria or microbe like pour plate method, streak plate method, swabbing by needle, pipetting, inoculation by loop, spreading by spreader, etc. After this, we see the bacterial growth after incubation of 24 hours, then according to the growth of bacteria antibiotics susceptibility test is conducted; this is done for sensitive antibiotics or resistance to that bacteria, and also for knowing the name of bacteria. Various methods like the dilution method, disk diffusion method, E test, etc., do antibiotics susceptibility tests. After that, various medicines are provided to the patients according to antibiotic sensitivity and resistance.

Keywords: inoculation, incubation, isolation, antibiotics suspectibility test, characterizing

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Authors: M. I. Ribah, M. Jibir, Y. A. Bashar, S. S. Manga

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Food safety is a significant and growing public health problem in the world and Nigeria as a developing country, since food-borne diseases are important contributors to the huge burden of sickness and death of humans. In Nigeria, traditional ready-to-eat meat products (RTE-MPs) like balangu, tsire, guru and dried meat products like kilishi, dambun nama, banda, were reported to be highly appreciated because of their eating qualities. The consumption of these products was considered as safe due to the treatments that are usually involved during their production process. However, during processing and handling, the products could be contaminated by pathogens that could cause food poisoning. Therefore, a hazard identification for pathogenic bacteria on some traditional RTE-MPs was conducted in Kebbi and Sokoto States, Nigeria. A total of 116 RTE-MPs (balangu-38, kilishi-39 and tsire-39) samples were obtained from retail outlets and analyzed using standard cultural microbiological procedures in general and selective enrichment media to isolate the target pathogens. A six-fold serial dilution was prepared and using the pour plating method, colonies were counted. Serial dilutions were selected based on the prepared pre-labeled Petri dishes for each sample. A volume of 10-12 ml of molten Nutrient agar cooled to 42-45°C was poured into each Petri dish and 1 ml each from dilutions of 102, 104 and 106 for every sample was respectively poured on a pre-labeled Petri plate after which colonies were counted. The isolated pathogens were identified and confirmed after series of biochemical tests. Frequencies and percentages were used to describe the presence of pathogens. The General Linear Model was used to analyze data on pathogen presence according to RTE-MPs and means were separated using the Tukey test at 0.05 confidence level. Of the 116 RTE-MPs samples collected, 35 (30.17%) samples were found to be contaminated with some tested pathogens. Prevalence results showed that Escherichia coli, salmonella and Staphylococcus aureus were present in the samples. Mean total bacterial count was 23.82×106 cfu/g. The frequency of individual pathogens isolated was; Staphylococcus aureus 18 (15.51%), Escherichia coli 12 (10.34%) and Salmonella 5 (4.31%). Also, among the RTE-MPs tested, the total bacterial counts were found to differ significantly (P < 0.05), with 1.81, 2.41 and 2.9×104 cfu/g for tsire, kilishi, and balangu, respectively. The study concluded that the presence of pathogenic bacteria in balangu could pose grave health risks to consumers, and hence, recommended good manufacturing practices in the production of balangu to improve the products’ safety.

Keywords: ready-to-eat meat products, retail outlets, public health, safety assessment

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Authors: Travis J. Meyer, Jasodra Ramlall, Phyo Thu, Nidhi Gadura

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For centuries humans have used the antimicrobial properties of copper to their advantage. Yet, after all these years the underlying mechanisms of copper mediated cell death in various microbes remain unclear. We had explored the hypothesis that copper mediated increased levels of lipid peroxidation in the membrane fatty acids is responsible for increased killing inEscherichia coli. In this study we show that in both gram positive (Staphylococcus aureus) and gram negative (Pseudomonas aeruginosa) bacteria there is a strong correlation between copper mediated cell death and increased levels of lipid peroxidation. Interestingly, the non-spore forming gram positive bacteria as well as gram negative bacteria show similar patterns of cell death, increased levels of lipid peroxidation, as well as genomic DNA degradation, however there is some difference inloss in membrane integrity upon exposure to copper alloy surface.

Keywords: antimicrobial, copper, gram positive, gram negative

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Authors: Bensaid Fatiha

Abstract:

The objective of our present work is the description of the antagonistic capacities of one strain of Pseudomonas fluorescens and the nonpathogenic fungic isolate Fusarium oxysporum against phytopathogenic agent Fusarium oxysporum F. Sp. lycopersici. This work has been achieved in two main parts: the first is interested on the in vitro antagonistic activities; the second was interested to study the soil receptiveness of fusarium wilt tomato. The use of strain of fluorescent Pseudomonas and a non-pathogenic strain of F. oxysporum in the different antagonism tests, has allowed assuring a certain bio-protection from the plants of tomatoes opposite to F. oxysporum F. Sp. lycopersici, agent of a wilt of tomato. These antagonistic have shown a substantial in vitro antagonistic activity on the three mediums (KB, PDA, KB+PDA) against F. oxysporum F. Sp. lycopersici, by inhibiting its growth mycelium with rate of inhibition going until 80 % with non-pathogen of Fusarium oxysporum and 60 % with strain of fluorescens Pseudomonas. Soil microbial balance, between the antagonistic population and that of pathogenic, can be modulated through microbiological variations or abiotic additives influencing directly or indirectly the metabolic behavior microbial. In this experiment, addition of glucose or EDTA, could increase or decrease the resistance of soil by activation of pathogenic or antagonists, as a result of modification and modulation in their metabolic activities.

Keywords: fluorescents, nonpathogenic, fusarium oxysporum, fusarium wilt, antagonism, biological control, soil receptivity

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Authors: Namratha Pai, K. Vasantharaj, K. Haribabu

Abstract:

Our Team is developing an innovative cost effective biological technique to desulfurize crude oil. ’Sulphur’ is found to be present in crude oil samples from .05% - 13.95% and its elimination by industrial methods is expensive currently. Materials required :- Alicyclobacillus acidoterrestrius, potato dextrose agar, oxygen, Pyragallol and inert gas(nitrogen). Method adapted and proposed:- 1) Growth of bacteria studied, energy needs. 2) Compatibility with crude-oil. 3) Reaction rate of bacteria studied and optimized. 4) Reaction development by computer simulation. 5) Simulated work tested by building the reactor. The method being developed requires the use of bacteria Alicyclobacillus acidoterrestrius - an acidothermophilic heterotrophic, soil dwelling aerobic, Sulfur bacteria. The bacteria are fed to crude oil in a unique manner. Its coated onto potato dextrose agar beads, cultured for 24 hours (growth time coincides with time when it begins reacting) and fed into the reactor. The beads are to be replenished with O2 by passing them through a jacket around the reactor which has O2 supply. The O2 can’t be supplied directly as crude oil is inflammable, hence the process. Beads are made to move around based on the concept of fluidized bed reactor. By controlling the velocity of inert gas pumped , the beads are made to settle down when exhausted of O2. It is recycled through the jacket where O2 is re-fed and beads which were inside the ring substitute the exhausted ones. Crude-oil is maintained between 1 atm-270 M Pa pressure and 45°C treated with tartaric acid (Ph reason for bacteria growth) for optimum output. Bacteria being of oxidising type react with Sulphur in crude-oil and liberate out SO4^2- and no gas. SO4^2- is absorbed into H2O. NaOH is fed once reaction is complete and beads separated. Crude-oil is thus separated of SO4^2-, thereby Sulphur, tartaric acid and other acids which are separated out. Bio-corrosion is taken care of by internal wall painting (phenolepoxy paints). Earlier methods used included use of Pseudomonas and Rhodococcus species. They were found to be inefficient, time and energy consuming and reduce the fuel value as they fed on skeleton.

Keywords: alicyclobacillus acidoterrestrius, potato dextrose agar, fluidized bed reactor principle, reaction time for bacteria, compatibility with crude oil

Procedia PDF Downloads 289

Authors: Abhichet Nobhiwong, Jiraporn Rojtinnakorn, Udomluk Sompong

Abstract:

Six probiotic strains isolated from Chiang Mai and Chiang Rai province, Thailand; CR1-2, CM3-4, CM5-2, CR7-8, CM10-5 and CM10-8 were used to study their morphology and inhibition activity on three pathogenic bacteria; Aeromonas sp., Streptococcus sp. and Flavobacterium sp. that isolated from infected Nile tilapia. The agar well diffusion technique was applied for 24 and 48 hours incubation. Interestingly, some probiotics showed good inhibition activity both 24 and 48 hours on each 3 bacterial pathogens. The capable inhibiting Aeromonas sp. were CR1-2 and CR5-2 with inhibition diameters of 13.0 mm and 11.2 mm, respectively. For Streptococcus sp., effective probiotics were CR10-2 with inhibition diameters of 10.7 mm. Whereas for Flavobacterium sp., effective probiotics were CR5-2 with inhibition diameter of 9.7 mm. It can be concluded that these probiotics have potentiality to develop as the pathogens biocontrol products. These will be support for safety and organic aquaculture that which the most worthy for people health.

Keywords: probiotics, Aeromanas sp., Streptococcus sp., Flavobacterium sp.

Procedia PDF Downloads 247

Authors: Yaling Gou, Sucai Yang, Pengwei Qiao

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Integrated chemical-biological treatment is an attractive alternative to remove polycyclic aromatic hydrocarbons (PAHs) from contaminated soil; wherein indigenous bacteria is the key factor for the biodegradation of residual PAHs concentrations after the application of chemical oxidation. However, the systematical study on the impact of persulfate (PS) oxidation on indigenous bacteria as well as PAHs removal is still scarce. In this study, the influences of different PS dosages (1%, 3%, 6%, and 10% [w/w]), as well as various activation methods (native iron, H2O2, alkaline, ferrous iron, and heat) on PAHs removal and indigenous bacteria in highly contaminated aged soil were investigated. Apparent degradation of PAHs in the soil treated with PS oxidation was observed, and the removal efficiency of total PAHs in the soil ranged from 38.28% to 79.97%. The removal efficiency of total PAHs in the soil increased with increasing consumption of PS. However, the bacterial abundance in soil was negatively affected following oxidation for all of the treatments added with PS, with bacterial abundance in the soil decreased by 0.89~2.88 orders of magnitude compared to the untreated soil. Moreover, the number of total bacteria in the soil decreased as PS consumption increased. Different PS activation methods and PS dosages exhibited different influences on the bacterial community composition. Bacteria capable of degrading PAHs under anoxic conditions were composed predominantly by Proteobacteria and Firmicutes. The total amount of Proteobacteria and Firmicutes also decreased with increasing consumption of PS. The results of this study provide important insight into the design of PAHs contaminated soil remediation projects.

Keywords: activation method, chemical oxidation, indigenous bacteria, polycyclic aromatic hydrocarbon

Procedia PDF Downloads 96

Authors: Abderrezzak khatib

Abstract:

Biogenic amines are low molecular weight nitrogenous compounds that have the potential to accumulate in food, posing a significant risk to food safety and human health. In this study, we investigated the inhibitory activity of three strains of lactic acid bacteria (LAB), against the growth and production of biogenic amines by both foodborne pathogens and food spoilage bacteria. The foodborne pathogens studied included Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella Paratyphi, while the food spoilage bacteria comprised Enterobacter cloacae and Proteus mirabilis. The methodology involved bacterial growth determination in petri dishes, bacterial culture extraction and derivatization, and biogenic amine analysis using HPLC. Our findings revealed that the inhibitory effects of LAB on these pathogens varied, with all three LAB strains demonstrating a remarkable reduction in the total bacterial count when combined with most pathogens, compared to the individual cultures of the pathogens. Furthermore, the presence of LAB in co-cultures with the pathogens resulted in a significant decrease in the production of tyramine and other biogenic amines by the pathogens themselves. These results suggest that LAB strains hold considerable promise in preventing the accumulation of biogenic amines in food products, thereby enhancing food safety. This study provides insights into the potential utilization of LAB in the context of preserving and ensuring the safety of food products. It highlights the significance of conducting additional research endeavors to elucidate the underlying mechanisms involved and to identify the precise bioactive compounds that are responsible for the observed inhibitory effects.

Keywords: food safety, lactic acid bacteria, foodborne pathogens, food spoilage bacteria, biogenic amines, tyrosine

Procedia PDF Downloads 23

Authors: D. Sengani, N. Potgieter, P. E. L. Mojapelo

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Three bacteria species which comprise of Gram negative and Gram positive microorganisms were isolated and identified on the basis of morpho-cultural study, catalase tests, oxidase tests and biochemical characteristics were found belonging to different genera including Burkholderia cepacia, Pasteurella pneumotropica and Enterococcus faecalis. The main objective of this study was to isolate and identify PCB degrading bacteria from PCB contaminated soils and test them for their degradation ability of PCBs in natural habitat conditions. The results indicated an overall decrease of PCB concentration level with the gradient average ranging from 1.5 to 1.8 respectively. Enterococcus faecalis removed as much as 32% of PCBs in the contaminated soil samples. Whereas Pasteurella pneumotropica could remove 24% of PCBs, Burkholderia cepacia 21% of PCBs and the mixed culture removed 23%. Data showed that the three bacterial strains could tolerate high concentration of PCBs. The results provided the evidence that naturally occurring bacteria in soil contaminated with PCBs have the potential to degrade PCBs. Statistical analysis showed that there was a significant positive correlation between bacteria growth and treatment with a coefficient of (r) =0.1459 and p value < 0.001.

Keywords: bacteria, bioaccumulation, biodegradation, bioremediation, polychlorinated biphenyls

Procedia PDF Downloads 219

Authors: Varangkana Thaotumpitak, Jarukorn Sripradite, Saharuetai Jeamsripong

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Environmental contamination from wastewater discharges originated from anthropogenic activities introduces the accumulation of enteropathogenic bacteria in aquatic animals, especially in oysters, and in shellfish harvesting areas. The consumption of raw or partially cooked oysters can be a risk for seafood-borne diseases in human. This study aimed to evaluate the relationship between the presence of Salmonella in oyster meat samples, and environmental factors (ambient air temperature, relative humidity, gust wind speed, average wind speed, tidal condition, precipitation and season) by using the principal component analysis (PCA). One hundred and forty-four oyster meat samples were collected from four oyster harvesting areas in Phang Nga province, Thailand from March 2016 to February 2017. The prevalence of Salmonella of each site was ranged from 25.0-36.11% in oyster meat. The results of PCA showed that ambient air temperature, relative humidity, and precipitation were main factors correlated with Salmonella detection in these oysters. Positive relationship was observed between positive Salmonella in the oysters and relative humidity (PC1=0.413) and precipitation (PC1=0.607), while the negative association was found between ambient air temperature (PC1=0.338) and the presence of Salmonella in oyster samples. These results suggested that lower temperature and higher precipitation and higher relative humidity will possibly effect on Salmonella contamination of oyster meat. During the high risk period, harvesting of oysters should be prohibited to reduce pathogenic bacteria contamination and to minimize a hazard of humans from Salmonellosis.

Keywords: oyster, Phang Nga Bay, principal component analysis, Salmonella

Procedia PDF Downloads 109

Authors: Maha A. Younus, Dhefaf H. Badri, Maha A. Al Abayaji, Taha M. Salih

Abstract:

Polymer metal complexes of poly vinyl alcohol and Cu (II), Ni (II), Mn (II) and Co (III) were prepared from the reaction of PVA with three different percentages of urea. The compound was characterized by fourier transform infrared spectrometry (FTIR) analysis and differential scanning calorimetric (DSC) Analysis. It has been established that the polymer and its metal complexes showed good activities against nine pathogenic bacteria (Escherichia coli, Klebsiellapneumonae, Staphylococcusaureus, Staphylococcus Albus, Salmonella Typhoid, Pseudomonas Aeruginosa, Shigella Dysentery, Proteus Morgani, Brucella Militensis). The polymer metal complexes show activity higher than that of the free polymer. The increasing activities were in the order (polymer < pol-Mn< pol-Co < pol-Ni ˂ pol-Cu). The ability of these compounds to show antimicrobial properties suggests that they can be further evaluated for medicinal and/or environmental applications.

Keywords: antimicrobial activity, PVA, polymer-metal complex, urea

Procedia PDF Downloads 310