Search results for: Clostridium botulinum

Authors: Mi-Hye Hwang, Young Min Son, Kichan Lee, Bang-Hun Hyun, Byeong Yeal Jung

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Botulism is a paralytic disease of human beings and animals caused by neurotoxin produced by Clostridium botulinum. The neurotoxins are genetically distinguished into 8 types, A to H. Ingestion of performed toxin, usually types B, C, and D, have been shown to produce diseases in most cases of cattle botulism. Vaccination is the best measure to prevent cattle botulism. However, the commercially available toxoid-based vaccines are difficult and hazardous to produce. We produced recombinant protein using gene of heavy chain domain of botulinum toxin B of which binds to cellular receptor of neuron cells and used as immunogen. In this study, we evaluated the safety and efficacy of botulism vaccine composed of recombinant types B. Safety test was done by National Regulation for Veterinary Biologicals. For efficacy test, female ICR mice (5 weeks old) were subcutaneously injected, intraperitoneally challenged, and examined the survival rates compared with vaccination and non-vaccination group. Mouse survival rate of recombinant types B vaccine was above 80%, while one of non-vaccination group was 0%. A vaccine composed of recombinant types B was safe and efficacious in mouse. Our results suggest that recombinant heavy chain receptor binding domain can be used as an effective vaccine candidate for type B botulism.

Keywords: botulism, livestock, vaccine, recombinant protein, toxin

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Authors: Nelson R. Osungbemiro, O. A. Bello-Olusoji, M. Oladipupo

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This study was carried out to determine the effects of probiotics Pseudomonas fluorescens on the growth performance, histology examination and immune modulation of African Catfish, (Clarias gariepinus) challenged with Clostridium botulinum. P. fluorescens, and C. botulinum isolates were removed from the gut, gill and skin organs of procured adult samples of Clarias gariepinus from commercial fish farms in Akure, Ondo State, Nigeria. The physical and biochemical tests were performed on the bacterial isolates using standard microbiological techniques for their identification. Antibacterial activity tests on P. fluorescens showed inhibition zone with mean value of 3.7 mm which indicates high level of antagonism. The experimental diets were prepared at different probiotics bacterial concentration comprises of five treatments of different bacterial suspension, including the control (T1), T2 (10³), T3 (10⁵), T4 (10⁷) and T5 (10⁹). Three replicates for each treatment type were prepared. Growth performance and nutrients utilization indices were calculated. The proximate analysis of fish carcass and experimental diet was carried out using standard methods. After feeding for 70 days, haematological values and histological test were done following standard methods; also a subgroup from each experimental treatment was challenged by inoculating Intraperitonieally (I/P) with different concentration of pathogenic C. botulinum. Statistically, there were significant differences (P < 0.05) in the growth performance and nutrient utilization of C. gariepinus. Best weight gain and feed conversion ratio were recorded in fish fed T4 (10⁷) and poorest value obtained in the control. Haematological analyses of C. gariepinus fed the experimental diets indicated that all the fish fed diets with P. fluorescens had marked significantly (p < 0.05) higher White Blood Cell than the control diet. The results of the challenge test showed that fish fed the control diet had the highest mortality rate. Histological examination of the gill, intestine, and liver of fish in this study showed several histopathological alterations in fish fed the control diets compared with those fed the P. fluorescens diets. The study indicated that the optimum level of P. fluorescens required for C. gariepinus growth and white blood cells formation is 10⁷ CFU g⁻¹, while carcass protein deposition required 10⁵ CFU g⁻¹ of P. fluorescens concentration. The study also confirmed P. fluorescens as efficient probiotics that is capable of improving the immune response of C. gariepinus against the attack of a virulent fish pathogen, C. botulinum.

Keywords: Clarias gariepinus, Clostridium botulinum, probiotics, Pseudomonas fluorescens

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Authors: Binh T. T. Nguyen, Zhenyu Li, Eric Yap, Yi Zhang, Ai-Qun Liu

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Silicon-photonic-sensor system is an emerging class of analytical technologies that use evanescent field wave to sensitively measure the slight difference in the surrounding environment. The wavelength shift induced by local refractive index change is used as an indicator in the system. These devices can be served as sensors for a wide variety of chemical or biomolecular detection in clinical and environmental fields. In our study, a system including a silicon-based micro-ring resonator, microfluidic channel, and optical processing is designed, fabricated for biomolecule detection. The system is demonstrated to detect Clostridium botulinum type A neurotoxin (BoNT) in different water sources. BoNT is one of the most toxic substances known and relatively easily obtained from a cultured bacteria source. The toxin is extremely lethal with LD50 of about 0.1µg/70kg intravenously, 1µg/ 70 kg by inhalation, and 70µg/kg orally. These factors make botulinum neurotoxins primary candidates as bioterrorism or biothreat agents. It is required to have a sensing system which can detect BoNT in a short time, high sensitive and automatic. For BoNT detection, silicon-based micro-ring resonator is modified with a linker for the immobilization of the anti-botulinum capture antibody. The enzymatic reaction is employed to increase the signal hence gains sensitivity. As a result, a detection limit to 30 pg/mL is achieved by our silicon-photonic sensor within a short period of 80 min. The sensor also shows high specificity versus the other type of botulinum. In the future, by designing the multifunctional waveguide array with fully automatic control system, it is simple to simultaneously detect multi-biomaterials at a low concentration within a short period. The system has a great potential to apply for online, real-time and high sensitivity for the label-free bimolecular rapid detection.

Keywords: biotoxin, photonic, ring resonator, sensor

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Authors: Alireza Moghtaderi, Negin Khakpour

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Hallux Valgus is a kind of Toes aberration where the Metatarsophalangeal joint that connects the big toe to the foot, leading to the inner side and a protrusion on the inner surface of toe arise. This study aimed to determine the effect of botulinum toxin A injection to reduce pain and deviation angle of the thumb in Hallux Valgus and to increase outcomes of treatment as an adjuvant therapy. Randomized clinical study was performed on 18 patients at the Clinic of Physical Medicine and Rehabilitation, Isfahan University of Medical Sciences. In this study the Halgvs valgus angle (HVA) between the metatarsals (IMA) and cartilage distal metatarsal angle (DMAA) and pain were assessed before and after injection. Average of Hallux Valgus angle before and after Botox injections were 28/89 ± 10/21 and 21/56 ± 8/22 degrees and the angle deviation in the 6 months after treatment was significantly improved (p <0.001). Injection of botulinum toxin A is a suitable and acceptable method to reform the skeleton deformities and also to reduce the pain in patients with Hallux valgus.

Keywords: metatasal, hallux valgus, pain, botulinum toxuin

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Authors: Ana Katrina C. Longos, Jarungchai Anton S. Vatanagul

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Objectives: To present a case of two Filipino male siblings initially seen with parkinsonism and eventually with dystonia and to present botulinum A toxin as part of the treatment for X-linked dystonia parkinsonism in Cebu City. Discussion: A 54 year old man presented initially with parkinsonian symptoms and later developed oromandibular and truncal dystonia. Further history revealed that he had an older brother who also presented with the same symptoms. Neuroimaging done on both patients revealed CVD infarcts in the pons and corona radiata respectively which where were not compatible with their symptoms. Family history revealed that their mother was originally from Panay and a diagnosis of X-linked dystonia parkinsonism (XDP) was made. Both patients were able to receive botulinum A toxin injections which provided temporary relief of symptoms. Conclusion: XDP was considered in 2 Filipino male siblings who presented with oromandibular dystonia, truncal dystonia, shuffling gait, resting tremors with ancestry from Panay on the maternal side. There is no cure for XDP, only symptomatic treatment. Until recently, only oral chemotherapy was available in Cebu. Botulinum A toxin injection done in both patients afforded temporary resolution of symptoms.

Keywords: XDP, dystonia of Panay, lubag, dystonia parkinsonism, botulinum a toxin

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Authors: Chen Zhang, Fengxue Xin, Jianzhong He

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A unique Clostridium beijerinckii species strain BGS1 was obtained from grass land samples, which is capable of producing 8.43g/L butanol and 3.21 isopropanol from 60g/L glucose while generating 4.68g/L volatile fatty acids (VFAs) from 30g/L xylan. The concentration of isopropanol produced by culture BGS1 is ~15% higher than previously reported wild-type Clostridium beijerinckii under similar conditions. Compared to traditional Acetone-Butanol-Ethanol (ABE) fermentation species, culture BGS1 only generates negligible amount of ethanol and acetone, but produces butanol and isopropanol as biosolvent end-products which are pure alcohols and more economical than ABE. More importantly, culture BGS1 can consume acetone to produce isopropanol, e.g., 1.84g/L isopropanol from 0.81g/L acetone in 60g/L glucose medium containing 6.15g/L acetone. The analysis of BGS1 draft genome annotated by RAST server demonstrates that no ethanol production is caused by the lack of pyruvate decarboxylase gene – related to ethanol production. In addition, an alcohol dehydrogenase (adhe gene) was found in BGS1 which could be a potential gene responsible for isopropanol-generation. This is the first report on Isopropanol-Butanol (IB) fermentation by wild-type Clostridium strain and its application for isopropanol and butanol production.

Keywords: acetone conversion, butanol, clostridium, isopropanol

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Authors: Sokol Isaraj, Lorela Bendo

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Botulinum toxin is the most commonly used treatment to reduce the appearance of dynamic facial wrinkles. It can smooth out wrinkles and restore a more youthful appearance. Although allergic reactions after botox injection are rare, care should be taken by the physician to diagnose the condition and provide suitable treatment in time. The authors report a case of allergic reaction with angioedema to abobotulinumtoxin A. A 50-year-old woman complaining of dynamic wrinkles was injected in a private clinic with Dysport. After two weeks, she returned to the clinic for the touch-up session. Thirty minutes after the completion of the injections in the crow’s feet area, she described the feeling of mild pain and warmth in the injected area, followed by angioedema. The symptoms couldn’t be controlled by IM corticosteroid, and the patient was referred to a hospital center. After adequate systemic treatment for four days, there was a resolution of the symptoms. Despite the reported safety of abobotulinumtoxin A, this case warns practitioners of unpredictably adverse reactions, which require rapid recognition and intravenous support.

Keywords: botulinum toxin, side effects, angioedema, injections

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Authors: A. Khalique, T. Naseem, N. Haque, Z. Rasool

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The objective of current study was to investigate the effect of Bacillus subtilis PB6 (CloSTAT) as a probiotic in broilers. The corn-soybean based diet was divided into four treatment groups; T1 (basal diet with no probiotic and no Clostridium perfringens); T2 (basal diet challenged with C. perfringens without probiotic); T3 (basal diet challenged with C. perfringens having 0.05% probiotic); T4 (basal diet challenged with C. perfringens having 0.1% probiotic). Every treatment group had four replicates with 24 birds each. Body weight and feed intake were measured on weekly basis, while ileal bacterial count was recorded on day-28 following Clostridium perfringens challenge. The 0.1% probiotic treatment showed 7.2% increase in average feed intake (P=0.05) and 8% increase in body weight compared to T2. In 0.1% treatment body weight was 5% higher than T3 (P=0.02). It was also observed that 0.1% treatment had improved feed conversion ratio (1.77) on 6^th week. No effect of treatment was observed on mortality and ileal bacterial count. The current study indicated that 0.1% use of probiotic had positive response in C. perfringens challenged broilers.

Keywords: Bacillus subtilis PB6, antibiotic growth promoters, Clostridium perfringens, broilers

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Authors: Diya Alsafadi, Fawwaz Khalili, Mohammad W. Amer

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Butanol is an important industrial solvent and potentially a better liquid transportation biofuel than ethanol. The cost of feedstock is one key problem associated with the biobutanol production. Date palm is sugar-rich fruit and highly abundant. Thousands of tons of date wastes that generated from date processing industries are thrown away each year and imposing serious environmental problems. To exploit the utilization of renewable biomass feedstock, date palm waste was utilized for butanol production by Clostridium acetobutylicum DSM 1731. Fermentation conditions were optimized by investigating several parameters that affect the production of butanol such as temperature, substrate concentration and pH. The highest butanol yield (1.0 g/L) and acetone, butanol, and ethanol (ABE) content (1.3 g/L) were achieved at 20 g/L date waste, pH 5.0 and 37 °C. These results suggest that date palm waste can be used for biobutanol production.

Keywords: biofuel, acetone-butanol-ethanol fermentation, date palm waste, Clostridium acetobutylicum

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Authors: Laura Cochrane

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The United States Centers for Disease Control and Prevention has established three categories of biological agents based on their ease of spread and the severity of the disease they cause. Category A biological agents are the highest priority because of their high degree of morbidity and mortality, ease of dissemination, the potential to cause social disruption and panic, special requirements for public health preparedness, and past use as a biological weapon. Despite the threat of Category A biological agents, opportunities for medical intervention exist. This work summarizes public information, consolidated and reviewed across the situational usefulness and disease awareness to offer discussion to three specific Category A agents: anthrax (Bacillus anthracis), botulism (Clostridium botulinum toxin), and smallpox (variola major), and provides an overview on the management of medical countermeasures available to treat these three (3) different types of pathogens. The medical countermeasures are discussed in the setting of pre-exposure prophylaxis, post-exposure prophylaxis, and therapeutic treatments to provide a framework for requirements in public health preparedness.

Keywords: anthrax, botulism, smallpox, medical countermeasures

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Authors: Nisha Singh, Munish Puri, Collin Barrow, Deepak Tuli, Anshu S. Mathur

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The biological conversion of lignocellulosic biomass to ethanol is a promising strategy to solve the present global crisis of exhausting fossil fuels. The existing bioethanol production technologies have cost constraints due to the involvement of mandate pretreatment and extensive enzyme production steps. A unique process configuration known as consolidated bioprocessing (CBP) is believed to be a potential cost-effective process due to its efficient integration of enzyme production, saccharification, and fermentation into one step. Due to several favorable reasons like single step conversion, no need of adding exogenous enzymes and facilitated product recovery, CBP has gained the attention of researchers worldwide. However, there are several technical and economic barriers which need to be overcome for making consolidated bioprocessing a commercially viable process. Finding a natural candidate CBP organism is critically important and thermophilic anaerobes are preferred microorganisms. The thermophilic anaerobes that can represent CBP mainly belong to genus Clostridium, Caldicellulosiruptor, Thermoanaerobacter, Thermoanaero bacterium, and Geobacillus etc. Amongst them, Clostridium thermocellum has received increased attention as a high utility CBP candidate due to its highest growth rate on crystalline cellulose, the presence of highly efficient cellulosome system and ability to produce ethanol directly from cellulose. Recently with the availability of genetic and molecular tools aiding the metabolic engineering of Clostridium thermocellum have further facilitated the viability of commercial CBP process. With this view, we have specifically screened cellulolytic and xylanolytic thermophilic anaerobic ethanol producing bacteria, from unexplored hot spring/s in India. One of the isolates is a potential CBP organism identified as a new strain of Clostridium thermocellum. This strain has shown superior avicel and xylan degradation under unoptimized conditions compared to reported wild type strains of Clostridium thermocellum and produced more than 50 mM ethanol in 72 hours from 1 % avicel at 60°C. Besides, this strain shows good ethanol tolerance and growth on both hexose and pentose sugars. Hence, with further optimization this new strain could be developed as a potential CBP microbe.

Keywords: Clostridium thermocellum, consolidated bioprocessing, ethanol, thermophilic anaerobes

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Authors: Mudassar Mohiuddin, Zahid Iqbal

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Introduction: Clostridium perfringens is an important pathogen responsible for causing enteric diseases in both human and animals. The bacteria produce several toxins. These toxins play vital role in the pathogenesis of various fatal enteric diseases and are classified into five types, on the basis of the differential production of Alpha, Beta, Epsilon and Iota toxins. In addition to the so-called major toxins, there are other toxins like beta2 toxin, produced by some strains of C. perfringens which may play a role in the pathogenesis of disease. Aim of the study: In this study a multiplex PCR assay was developed and used for detection of cpb2 gene to identify the Beta2 harboring isolates among different types of C. perfringens. Objectives: The primary objective of this study was to identify the prevalence of β2-toxin gene in local isolates of Clostridium perfringens. Methodology: This was an experimental study. Random sampling technique was used. A total of 97 sheep and goats were included in this study. All were Pakistani local breeds. The samples were collected during the period from Sep, 2014 to Mar, 2015 from selected districts of Punjab province (Pakistan). Faecal samples were cultured in cooked meat media. The identification of Clostridium perfringens was made on the basis of biochemical tests. Multiplex PCR was performed to identify the toxin genes. Results: A total of 43 C. perfringens isolates were genotyped using multiplex PCR assay. The gene encoding C. perfringens β2-toxin (cpb2) was present in more than 50% of the isolates genotyped. However, the prevalence of this gene varied between sheep and goat isolates. Conclusion: The present study suggests the high occurrence of C. perfringens b2-toxin (cpb2) in the local isolates of Pakistan. As β2-toxin is present in both healthy and diseased animals, so further studies are suggested to establish the role of β2-toxin in pathogenesis of the clostridial enteric diseases.

Keywords: beta 2 toxin gene, clostridium perfringens, enteric diseases, goats, multiplex PCR, sheep

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Authors: Sadhana Deshmukh

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Objective: Men represent only a fraction of the medical aesthetic practice. They are increasingly becoming more cosmetically-inclined. The primary objective is to harmonize facial proportion by prioritizing and focusing on forehead nose, cheek and chin complex. Introduction: Despite tremendous variability, diverse population of the Indian subcontinent, the male skull is unique in its overall larger size, and shape. Men tend to have a large forehead with prominent supraorbital ridges, wide glabella, square orbit, and a prominent protruding mandible. Men have increased skeletal muscle mass, with less facial subcutaneous fat. Facial aesthetics is evolving rapidly. Commonly published canons of facial proportions usually represent feminine standards and are not applicable to males. Strict adherence to these norms is therefore not necessary to obtain satisfying results in male patients. Materials and Methods: Male patients age group 30-60 years have been enrolled. Botulinum toxin and hyaluronic acid fillers were used to update consensus recommendations for facial rejuvenation using these two types of products alone and in combination. Results: There are specific recommendations by facial area, focusing on relaxing musculature, restoring volume, recontouring using toxin and dermal fillers alone and in combination. For upper face, though botulinum toxin remains the cornerstone of treatment, temples and forehead fillers are recommended for optimal results. In Mid face, these fillers are placed more laterally to maintain the masculine look. Botulinum toxin and fillers in combination can improve outcomes in the lower face. Chin augmentation remains the center point for lower face. Conclusions: Males are more likely to have shorter doctor visits, less likely to ask questions, have a lower attention to bodily changes. The physician must patiently gauge male patients’ aging and cosmetic goals. Clinicians can also benefit from ongoing guidance on products, tailoring treatments, treating multiple facial areas, and using combinations of products. An appreciation that rejuvenation is 3-dimensional process involving muscle control, volume restoration and recontouring helps.

Keywords: male aesthetics, botulinum toxin, hyaluronic acid dermal fillers, Asian patients

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Authors: Akulov M. A., Orlova O. R., Zaharov V. O., Tomskij A. A.

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Introduction: One of the common postoperative complications of posterior cranial fossa (PCF) and cerebello-pontine angle tumor treatment is a facial nerve palsy, which leads to multiple and resistant to treatment impairments of mimic muscles structure and functions. After 4-6 months after facial nerve palsy with insufficient therapeutic intervention patients develop a postparalythic syndrome, which includes such symptoms as mimic muscle insufficiency, mimic muscle contractures, synkinesis and spontaneous muscular twitching. A novel method of treatment is the use of a recent local neuromuscular blocking agent– botulinum toxin A (BTA). Experience of BTA treatment enables an assumption that it can be successfully used in late facial nerve palsy complications to significantly increase quality of life of patients. Study aim. To evaluate the efficacy of botulinum toxin A (BTA) (Xeomin) treatment in patients with late facial nerve palsy complications. Patients and Methods: 31 patients aged 27-59 years 6 months after facial nerve palsy development were evaluated. All patients received conventional treatment, including massage, movement therapy etc. Facial nerve palsy developed after acoustic nerve tumor resection in 23 (74,2%) patients, petroclival meningioma resection – in 8 (25,8%) patients. The first group included 17 (54,8%) patients, receiving BT-therapy; the second group – 14 (45,2%) patients continuing conventional treatment. BT-injections were performed in synkinesis or contracture points 1-2 U on injured site and 2-4 U on healthy side (for symmetry). Facial nerve function was evaluated on 2 and 4 months of therapy according to House-Brackman scale. Pain syndrome alleviation was assessed on VAS. Results: At baseline all patients in the first and second groups demonstrated аpostparalytic syndrome. We observed a significant improvement in patients receiving BTA after only one month of treatment. Mean VAS score at baseline was 80,4±18,7 and 77,9±18,2 in the first and second group, respectively. In the first group after one month of treatment we observed a significant decrease of pain syndrome – mean VAS score was 44,7±10,2 (р<0,01), whereas in the second group VAS score was as high as 61,8±9,4 points (p>0,05). By the 3d month of treatment pain syndrome intensity continued to decrease in both groups, but, the first group demonstrated significantly better results; mean score was 8,2±3,1 and 31,8±4,6 in the first and second group, respectively (р<0,01). Total House-Brackman score at baseline was 3,67±0,16 in the first group and 3,74±0,19 in the second group. Treatment resulted in a significant symptom improvement in the first group, with no improvement in the second group. After 4 months of treatment House-Brockman score in the first group was 3,1-fold lower, than in the second group (р<0,05). Conclusion: Botulinum toxin injections decrease postparalytic syndrome symptoms in patients with facial nerve palsy.

Keywords: botulinum toxin, facial nerve palsy, postparalytic syndrome, synkinesis

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Authors: Akulov M. A., Zaharov V. O., Jurishhev P. E., Tomskij A. A.

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Introduction: Spasticity is a severe consequence of stroke, leading to profound disability, decreased quality of life and decrease of rehabilitation efficacy [4]. Spasticity is often associated with pain syndrome, arising from joint damage of paretic limbs (postural arthropathy) or painful spasm of paretic limb muscles. It is generally accepted that injection of botulinum toxin into a cramped muscle leads to decrease of muscle tone and improves motion range in paretic limb, which is accompanied by pain alleviation. Study aim: To evaluate the change in pain syndrome intensity after incections of botulinum toxin A (Xeomin) in stroke patients with upper limb spasticity. Patients and methods. 21 patients aged 47-74 years were evaluated. Inclusion criteria were: acute stroke 4-7 months before the inclusion into the study, leading to spasticity of wrist and/or finger flexors, elbow flexor or forearm pronator, associated with severe pain syndrome. Patients received Xeomin as monotherapy 90-300 U, according to spasticity pattern. Efficacy evaluation was performed using Ashworth scale, disability assessment scale (DAS), caregiver burden scale and global treatment benefit assessment on weeks 2, 4, 8 and 12. Efficacy criterion was the decrease of pain syndrome by week 4 on PQLS and VAS. Results: The study revealed a significant improvement of measured indices after 4 weeks of treatment, which persisted until the 12 week of treatment. Xeomin is effective in reducing muscle tone of flexors of wrist, fingers and elbow, forearm pronators. By the 4th week of treatment we observed a significant improvement on DAS (р < 0,05), Ashworth scale (1-2 points) in all patients (р < 0,05), caregiver burden scale (р < 0,05). A significant decrease of pain syndrome by the 4th week of treatment on PQLS (р < 0,05) и VAS (р < 0,05) was observed. No adverse effect were registered. Conclusion: Xeomin is an effective treatment of pain syndrome in postural upper limb spasticity after stroke. Xeomin treatment leads to a significant improvement on PQLS and VAS.

Keywords: botulinum toxin, pain syndrome, spasticity, stroke

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Authors: Min-Shiuan Liou, Yi Shan Yang, Yang-Zhan Huang, Chia-Wen Hsieh

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A high speed growing and butanol-tolerant Clostridium acetobutylicum HOL1 mutant was screened throughout continuous adaption culture with C. acetobutylicum ATCC 824. The HOL1 strain can grow well in 10 g/L butanol contained CGM medium and can produce about 12.8 g /L butanol during 24 hrs. The C. acetobutylicum HOL1 strain was able to produce 166 mM butanol with 21 mM acetone at pH 4.8, resulting in a butanol selectivity (a molar ratio of butanol to total solvents) of 0.79, which is much higher than that (0.6) of the wild-type strain C. acetobutylicum ATCC 824. The acetate and butyrate accumulation were not observed during fermentation of the HOL1 strain. A hyper-butanol producing C. acetobutylicum HOL1 (pBPHS-3), which was created to overexpress the Bacillus psychrosaccharolyticus originated specific heat-shock protein gene, hspX, from a clostridial phosphotransbutyrylase promoter, was studied for its potential to produce a high titer of butanol. Overexpression of hspX resulted in increased final butanol yield 47% and 30% higher than those of the the ATCC824 and the HOL1 strains, respectively. The remarkable high-speed growth and butanol tolerance of strain HOL1 (pBPHS-3) demonstrates that overexpression of heterogeneous stress protein-encoding gene, hspX, could help C. acetobutylicum to effectively produce a high concentration of butanol.

Keywords: Clostridium acetobutylicum, butanol, heat-shock protein, resistance

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Authors: Jihane Saad, Thomas Lendormi, Caroline Le Marechal, Anne-marie Pourcher, Céline Druilhe, Jean-louis Lanoiselle

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Agricultural anaerobic digestion consists in the conversion of animal slurry and manure into biogas and digestate. They need, however, to be treated at 70 ºC during 60 min before anaerobic digestion according to the European regulation (EC n°1069/2009 & EU n°142/2011). The impact of such heat treatment on the outcome of bacteria has been poorly studied up to now. Moreover, a recent study¹ has shown that enterococci and clostridia are still detected despite the application of such thermal treatment, questioning the relevance of this approach for the hygienisation of digestate. The aim of this study is to establish the heat inactivation kinetics of two species of enterococci (Enterococcus faecalis and Enterococcus faecium) and two species of clostridia (Clostridioides difficile and Clostridium novyi as a non-toxic model for Clostridium botulinum of group III). A pure culture of each strain was prepared in a specific sterile medium at concentration of 10⁴ – 10⁷ MPN / mL (Most Probable number), depending on the bacterial species. Bacterial suspensions were then filled in sterilized capillary tubes and placed in a water or oil bath at desired temperature for a specific period of time. Each bacterial suspension was enumerated using a MPN approach, and tests were repeated three times for each temperature/time couple. The inactivation kinetics of the four indicator bacteria is described using the Weibull model and the classical Bigelow model of first-order kinetics. The Weibull model takes biological variation, with respect to thermal inactivation, into account and is basically a statistical model of distribution of inactivation times as the classical first-order approach is a special case of the Weibull model. The heat treatment at 70 ºC / 60 min contributes to a reduction greater than 5 log10 for E. faecium and E. faecalis. However, it results only in a reduction of about 0.7 log10 for C. difficile and an increase of 0.5 log10 for C. novyi. Application of treatments at higher temperatures is required to reach a reduction greater or equal to 3 log10 for C. novyi (such as 30 min / 100 ºC, 13 min / 105 ºC, 3 min / 110 ºC, and 1 min / 115 ºC), raising the question of the relevance of the application of heat treatment at 70 ºC / 60 min for these spore-forming bacteria. To conclude, the heat treatment (70 ºC / 60 min) defined by the European regulation is sufficient to inactivate non-sporulating bacteria. Higher temperatures (> 100 ºC) are required as far as spore-forming bacteria concerns to reach a 3 log10 reduction (sporicidal activity).

Keywords: heat treatment, enterococci, clostridia, inactivation kinetics

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Authors: Ashana Gupta

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Aim: In the UK, Botulinum Toxin (botox) is authorised for treating chronic myofascial pain secondary to masseter muscle hypertrophy (Fedorowicz et al. 2013). This audit aimed to ensure the Maxillofacial Unit is meeting the trust guidelines for the safe storage and disposal of botox. Method: The trust upholds a strict policy for botox handling. The audit was designed to optimise several elements including Staff awareness of regulations around botox handling A questionnaire was designed to test knowledge of advised storage temperatures, reporting of adverse events, disposal procedures and regulatory authorities. Steps taken to safely delivertoxin and eliminate unused toxin. A checklist was completed. These include marks for storagetemperature, identification checks, disposal of sharps, deactivation of toxin, and disposal. Results: All staff correctly stated storage requirements for toxin. 75% staff (n=8) were unsure about reporting and regulations. Whilst all staff knew how to dispose of vials, 0% staff showed awareness for the crucial step of deactivating toxin. All checklists (n=20) scored 100% for adequate storage, ID checks, and toxin disposal. However, there were no steps taken to deactivate toxin in any cases. Staff training took place with revision to clinical protocols. In line with Trust guidelines, an additional clinical step has been introduced including use of 0.5% sodium hypochlorite to deactivate botox. Conclusion: Deactivation is crucial to ensure residual toxin is not misused. There are cases of stolen botox within South-Tees Hospital (Woodcock, 2014). This audit was successful in increasing compliance to safe handling and disposal of botox by 100% and ensured our hospitalmeets Trust guidance.

Keywords: botulinum toxin, aesthetics, handling, disposal

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Authors: Danijela Vuković, Radovan Čobanović, Milorad Plačkić

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The expansion of population imposes increase in usage of animal meat, on whose quality directly affects the quality of the feed that the animals are fed with. The selection of raw materials, hygiene during the technological process, various hydrothermal treatments, methods of mixing etc. have an influence on the quality of feed. Monitoring of the feed is very important to obtain information about the quality of feed and the possible prevention of animal diseases which can lead to different human diseases outbreaks. In this study parameters of feed safety were monitored. According to the mentioned, the goal of this study was to evaluate microbiological safety of feed (feedstuffs and complete mixtures). Total number of analyzed samples was 4399. Analyzed feed samples were collected in various retail shops and feed factories during the period of 44 months (from January 2013 untill September 2017). Samples were analyzed on Salmonella spp. and Clostridium perfringens in quantity of 50g according to Serbian regulation. All microorganisms were tested according to ISO methodology: Salmonella spp. ISO 6579:2002 and Clostridium perfringens ISO 7937:2004. Out of 4399 analyzed feed samples 97,5% were satisfactory and 2,5% unsatisfactory concerning Salmonella spp. As far as Clostridium perfringens is concerned 100% of analyzed samples were satisfactory. The obtained results suggest that technological processing of feed in Serbia is at high level when it comes to safety and hygiene of the products, but there are still possibilities for progress and improvement which only can be reached trough the permanent monitoring of feed.

Keywords: microbiology, safety, hygiene, feed

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Authors: Jawaher F. Alsharef, Abdullah A. Ghaddaf, Mohammed S. Alomari, Abdullah A. Al Qurashi, Ahmed S. Abdulhamid, Mohammed S. Alshehri, Majed Alosaimi

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Botulinum toxin type A (BTX-A) is the most popular therapeutic agent for muscle relaxation and pain control. Lately, BTX-A injection received great interest as a part of multimodal pain management for lower limb lengthening and deformity correction. This systematic review aimed to determine the role of BTX-A injection in pain management for during lower limb lengthening and/or deformity correction. We searched Medline, Embase, and CENTRAL. We included randomized controlled trials (RCTs) that compared the BTX-A injection to placebo for individuals undergoing lower limb lengthening and/or deformity correction. We sought to evaluate the following outcomes: pain on visual analogue scale (VAS), range of motion parameters, average opioid consumption, and adverse events. The standardized mean difference (SMD) was used to represent continuous outcomes while risk ratio (RR) was used to represent dichotomous outcomes. A total of 4 RCTs that enrolled 257 participants (337 limbs) deemed eligible. Adjuvant BTX-A injection showed a significant reduction in post-operative pain compared to placebo (SMD=–0.28, 95% CI –0.53 to –0.04). No difference was found between BTX-A injection and placebo in terms of range of motion parameters, average opioid consumption, or adverse events after surgical limb lengthening and/or deformity correction (RR= 0.77, 95% CI –0.58 to 1.03). Conclusions: Adjuvant BTX-A injection conferred a discernible reduction in post-operative pain during surgical limb lengthening and/or deformity without increasing the risk of adverse events.

Keywords: botulinum toxin type A, limb lengthening, distraction osteogenesis, deformity correction, pain management

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Authors: Daniela Prado, Lexi Frankel, Amalia Ardeljan, Lokesh Manjani, Matthew Cardeiro, Omar Rashid

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Background: Clostridium difficile (C. diff) is a gram-positive bacterium that is known to cause life-threatening diarrhea and severe inflammation of the colon. It originates as an alteration of the gut microbiome and can be transmitted through spores. Recent studies have shown a high association between the development of C. diff in cancer patients due to extensive hospitalization. However, research is lacking regarding C. diff’s association in the causation or prevention of cancer. The objective of this study was to therefore assess the correlation between Clostridium difficile infection (CDI) and the incidence of bone cancer. Methods: This retrospective analysis used data provided by a Health Insurance Portability and Accountability Act (HIPAA) compliant national database to evaluate the patients infected versus patients not infected with C. diff using ICD-10 and ICD-9 codes. Access to the database was granted by the Holy Cross Health, Fort Lauderdale, for the purpose of academic research. Standard statistical methods were used. Results: Between January 2010 and December 2019, the query was analyzed and resulted in 78863 patients in both the infected and control group, respectively. The two groups were matched by age range and CCI score. The incidence of bone cancer was 659 patients (0.835%) in the C. diff group compared to 1941 patients (2.461%) in the control group. The difference was statistically significant by a P-value < 2.2x10^-16 with an odds ratio (OR)= 0.33 (0.31-0.37) with a 95% confidence interval (CI). Treatment for CDI was analyzed for both C. diff infected and noninfected populations. 91 out of 16,676 (0.55%) patients with a prior C. diff infection and treated with antibiotics were compared to the control group were 275 out of 16,676 (1.65%) patients with no history of CDI and received antibiotic treatment. Results remained statistically significant by P-value <2.2x10-16 with an OR= 0.42 (0.37, 0.48). and a 95% CI. Conclusion: The study shows a statistically significant correlation between C. diff and a reduced incidence of bone cancer. Further evaluation is recommended to assess the potential of C. difficile in reducing bone cancer incidence.

Keywords: bone cancer, colitis, clostridium difficile, microbiome

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Authors: Ga-Young Lee, Hyun-Man Kim

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The modulation of the cell-cell junction is critical in epithelial-mesenchymal transition during tumorigenesis. As RhoA activity is known to be up-regulated to dissociate cell-cell junction by contracting acto-myosin complex in various cancer cells, the present study investigated if RhoA activity was also associated with the disruption of the cell-cell junction of oral cancer cells. We studied SCC-25 cells which are established from oral squamous cell carcinoma if their E-cadherin junction (ECJ) was under control of RhoA. Interestingly, development of ECJ of SCC-25 cells depended on the amount of fibronectin (FN) coated on the culture dishes. Seeded cells promptly aggregated to develop ECJ on the substrates coated with a low amount of FN, whereas they were retarded in the development of ECJ on the substrates coated with a high amount of FN. However, it was an unexpected finding that total RhoA activity was lower in the dissociated cells on the substrates of high FN than in the aggregated cells on the substrates of low FN. Treating the dissociated cells on the substrates of high FN with LPA, a RhoA activator, promoted the development to ECJ. In contrast, treating the aggregated cells on the substrates of low FN with Clostridium botulinum C3, a toxin decreasing RhoA activity, dissociated cells concomitant with the disruption of ECJ. Genetical knockdown of RhoA expression by transfecting RhoA siRNA also down-regulated the development of ECJ in SCC-25 cells. Furthermore, PMA, an activator of protein kinase C (PKC), down-regulated the development of ECJ junction of SCC-25 cells on the substrates coated with low FN. In contrast, GO6976, a PKC inhibitor, up-regulated the development of ECJ of SCC-25 cells with the activation of RhoA on the substrates coated with high FN. In conclusion, in the present study, we demonstrated unexpected results that the activation of RhoA promotes the development of ECJ, whereas the inhibition of RhoA retards the development of ECJ in SCC-25 cells.

Keywords: E-cadherin junction, oral squamous cell carcinoma, PKC, RhoA, SCC-25

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Authors: Jessica Pinheiro Silva, Brenda Rabello De Camargo, Daniel Gusmao De Morais, Eliane Ferreira Noronha

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The utilization of lignocellulosic biomass as source of polysaccharides for industrial applications requires an arsenal of enzymes with different mode of action able to hydrolyze its complex and recalcitrant structure. Clostridium thermocellum is gram-positive, thermophilic bacterium producing lignocellulosic hydrolyzing enzymes in the form of multi-enzyme complex, termed celulossomes. This complex has several hydrolytic enzymes attached to a large and enzymically inactive protein known as Cellulosome-integrating protein (CipA), which serves as a scaffolding protein for the complex produced. This attachment occurs through specific interactions between cohesin modules of CipA and dockerin modules in enzymes. The present work aims to construct celulosomes in vitro with the structural protein CipA, a xylanase called Xyn10D and a cellulose called CelJ from C.thermocellum. A mini-scafoldin was constructed from modules derived from CipA containing two cohesion modules. This was cloned and expressed in Escherichia coli. The other two genes were cloned under the control of the alcohol oxidase 1 promoter (AOX1) in the vector pPIC9 and integrated into the genome of the methylotrophic yeast Pichia pastoris GS115. Purification of each protein is being carried out. Further studies regarding enzymatic activity of the cellulosome is going to be evaluated. The cellulosome built in vitro and composed of mini-CipA, CelJ and Xyn10D, can be very interesting for application in industrial processes involving the degradation of plant biomass.

Keywords: cellulosome, CipA, Clostridium thermocellum, cohesin, dockerin, yeast

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Authors: Karla Cautivo, Thomas Riley, Daniel Knight

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Clostridium difficile infection (CDI) is the most common cause of infectious diarrhea in hospitalised humans. C. difficile colonises the gastrointestinal tract, causes disease in a variety of animal species and can persist as a spore in diverse environments. Genetic overlap between C. difficile strains from human, animal and environmental sources suggests CDI has a zoonotic or foodborne aetiology. In Australia, C. difficile PCR ribotype RT014 (MLST clade 1) and several ST11 (MLST clade 5) RTs are found commonly in livestock. The high prevalence and diversity of ST11 strains in Australian production animals indicates Australia might be the ancestral home for this lineage. This project describes for the first time the ecology of C. difficile in Australian native animals, providing insights into the prevalence, molecular epidemiology and evolution of C. difficile in this unique environment and a possible role in CDI in humans and animals in Australia. Faecal samples were collected from wild/captive reptiles (n=37), mammals (n=104) and birds (n=102) in Western Australia in 2020/21. Anaerobic enrichment culture was performed, and C. difficile isolates were characterised by PCR ribotyping and toxin gene profiling. Seventy isolates of C. difficile were recovered (prevalence of C. difficile in faecal samples 28%, n=68/243); 27 unique RTs were identified, 5 were novel. The prevalence of C. difficile was similar for reptiles and mammals, 46% (n=17/37) and 43%(n=45/104), respectively, but significantly lower in birds (7.8%, n=8/102; p<0.00001 for both reptiles and mammals). Of the 57 isolates available for typing, RT237 (clade 5) and RT002 (clade 2) were the most prevalent, 15.8% (n=9/57) and 14% (n=8/57), respectively. The high prevalence of C. difficile in reptiles and mammals, particularly clade 5 strains, supported by previous studies of C. difficile in Australian soils, suggest that Australia might be the ancestral home of MLST clade 5.

Keywords: Clostridium difficile, zoonosis, molecular epidemiology, ecology and evolution

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Authors: Ni Luh Putu Harta Wedari

Abstract:

Background: Clostridium difficile has two main virulence factors, namely TcdA and TcdB. TcdA encoded by the tcdA gene acts as an enterotoxin, pro-inflammatory and fluid accumulation, while TcdB encoded by the tcdB gene is cytotoxic, causes disruption of the actin cytoskeleton, and causes disruption of tight junctions in colon cells. This study aims to explore the relationship between the tcdA and tcdB genes and the duration of diarrhea in elderly patients. Method: This research was an observational analytic with a prospective cross-sectional with samples of elderly diarrhea patients who met the inclusion criteria in Denpasar City health service facilities from 1 December 2022 until 30 June 2023, and then their feces were analyzed using the real-time PCR method. Results: In this study, 40 elderly diarrhea patients met the inclusion criteria and in accordance with the minimum sample size, 28 (70%) men and 12 (30%) women. 5 patients (12.5%) had a history of azithromycin, 4 (10%) levofloxacin, 17 (42.5%) ciprofloxacin, 8 (20%) metronidazole, 1 (2.5%) cefoperazone, 5 (12, 5%) doxycycline. Comorbids, namely 13 (32.5%) type II diabetes mellitus, 4 (10%) chronic kidney disease, 10 (25%) malignancies, 7 (17.5%) urinary tract infections, 3 (7.5%) %) immunocompromised, 2 (5%) cardiac heart failure, and 1 (2.5%) acute on chronic kidney disease. The overall diarrhea duration average was 5 days. 8 samples (20%) were positive for 16s rRNA, and there was no significant difference in diarrhea duration with negative samples (p=0.166). The relationship between the tcdA gene and the duration of diarrhea could not be performed because all samples were negative. Likewise, relationship analysis between the coexistence of tcdA and tcdB could not be performed. There was no significant difference between tcdB positive 3 (7.5%) and negative with diarrhea duration (p=0.739). Conclusion: There is no significant relationship between the presence of the 16s rRNA and tcdB C. difficile genes with the duration of diarrhea in elderly patients.

Keywords: clostridium, difficile, diarrhea, elderly, tcdA, tcdB

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Authors: Carlos A. Garcia-Mogollon, Juan C. Quintero-Diaz, Claudio Avignone-Rossa

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Clostridium saccharoperbutylacetonicum 1-4N (Csb 1-4N) is an industrial reference strain for Acetone-Butanol-Ethanol (ABE) fermentation. Csb 1-4N is a solventogenic clostridium and H₂ producer with a metabolic profile that makes it a good candidate for Bioelectrosynthesis System (BES). The aim of this study was to evaluate the electroactivity of Csb 1-4N by cyclic voltammetry technique (CV). The Bioelectrosynthesis fermentation (BES) started in a Triptone-Yeast extract (TY) medium with trace elements and vitamins, Complex Nitrogen Source (CNS), and bicarbonate (NaHCO₃, 4g/L) as a carbon source, run at -600mVAg/AgCl and adding 200uM NADH. The six BES batches were performed with different media composition with and without NADH, CNS, HCO₃⁻ , and applied potential. The CV was performed as three-electrode system: platinum slice working electrode (WE), nickel contra electrode (CE) and reference electrode Ag/AgCl (ER). CVs were run in a potential range of -0.7V to 0.7V vs. VAg/AgCl at a scan rate 10mV/s. A CV recorded using different NaHCO₃ concentrations (0.25; 0.5; 1.0; 4g/L) were obtained. BES fermentation samples were centrifuged (3000 rpm, 5min, 4C), and supernatant (7mL) was used. CVs were obtained for Csb1-4N BES culture cell-free supernatant at 0h, 24h, and 48h. The electrochemical analysis was carried out with a PalmSens 4.0 potentiostat/galvanostat controlled with the PStrace 5.7 software, and CVs curves were characterized by reduction and oxidation currents and reduction and oxidation peaks. The CVs obtained for NaHCO₃ solutions showed that the reduction current and oxidation current decreased as the NaHCO₃ concentration was decreased. All reduction and oxidation currents decreased until exponential growth stop (24h), independence of initial cathodic current, except in medium with trace elements, vitamins, and NaHCO3, in which reduction current was around half at 24h and followed decreasing at 48. In this medium, Csb1-4N did not grow, but pH was increased, indicating that NaHCO₃ was reduced as the reduction current decreased. In general, at 48h reduction currents did not present important changes between different mediums in BES cultures. In terms of intensities in the peaks (Ip) did not present important variations; except with Ipa and Ipc in BES culture with NaHCO₃ and NADH added are higher than peaks in other cultures. Based on results, cathodic and anodic currents changes were induced by NaHCO₃ reduction reactions during Csb1-4N metabolic activity in different BES experiments.

Keywords: clostridium saccharoperbutylacetonicum 1-4N, bioelectrosynthesis, carbon dioxide fixation, cyclic voltammetry

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Authors: Rajalaxmi N., Padma Bhat, Pooja Garag, Pooja N. M., V. S. Hombalimath

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Microbial fuel cell (MFC) is the advancement of science that aims at utilizing the oxidizing potential of bacteria for wastewater treatment and production of bio-hydrogen and bio-electricity. Salt-bridge is the economic alternative to highly priced proton-exchange membrane in the construction of a microbial fuel cell. This paper studies the electricity generating capacity of E.coli and Clostridium sporogenes in microbial fuel cells (MFCs). Unlike most of MFC research, this targets the long term goals of renewable energy production and wastewater treatment. In present study the feasibility and potential of bioelectricity production from different wastewater was observed. Different wastewater was primarily treated which were confirmed by the COD tests which showed reduction of COD. We observe that the electricity production of MFCs decreases almost linearly after 120 hrs. The sewage wastewater containing Clostridium sporogenes showed bioelectricity production up to 188mV with COD removal of 60.52%. Sewage wastewater efficiently produces bioelectricity and this also helpful to reduce wastewater pollution load.

Keywords: microbial fuel cell, bioelectricity, wastewater, salt bridge, COD

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Authors: Aparna Lal, Ashwin Swaminathan, Teisa Holani

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Background: Clostridium difficile infections (CDIs) have been on the rise globally. In Australia, rates of CDI in all States and Territories have increased significantly since mid-2011. Identifying risk factors for CDI in the community can help inform targeted interventions to reduce infection. Methods: We examine the role of neighbourhood socio-economic status, demography, testing practices and the number of residential aged care facilities on spatial patterns in CDI incidence in the Australian Capital Territory. Data on all tests conducted for CDI were obtained from ACT Pathology by postcode for the period 1st January 2004 through 31 December 2014. Distribution of age groups and the neighbourhood Index of Relative Socio-economic Advantage Disadvantage (IRSAD) were obtained from the Australian Bureau of Statistics 2011 National Census data. A Bayesian spatial conditional autoregressive model was fitted at the postcode level to quantify the relationship between CDI and socio-demographic factors. To identify CDI hotspots, exceedance probabilities were set at a threshold of twice the estimated relative risk. Results: CDI showed a positive spatial association with the number of tests (RR=1.01, 95% CI 1.00, 1.02) and the resident population over 65 years (RR=1.00, 95% CI 1.00, 1.01). The standardized index of relative socio-economic advantage disadvantage (IRSAD) was significantly negatively associated with CDI (RR=0.74, 95% CI 0.56, 0.94). We identified three postcodes with high probability (0.8-1.0) of excess risk. Conclusions: Here, we demonstrate geographic variations in CDI in the ACT with a positive association of CDI with socioeconomic disadvantage and identify areas with a high probability of elevated risk compared with surrounding communities. These findings highlight community-based risk factors for CDI.

Keywords: spatial, socio-demographic, infection, Clostridium difficile

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Authors: Asma Afshari, Abdollah Jamshidi, Jamshid Razmyar, Mehrnaz Rad

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Clostridium perfringens is a serious pathogen which causes enteric diseases in domestic animals and food poisoning in humans. Spores can survive cooking processes and play an important role in the possible onset of disease. In this study RAPD-PCR and REP-PCR were used to examine the genetic diversity of 49isolates ofC. Perfringens type A from 3 different sources. The results of RAPD-PCR revealed the most genetic diversity among poultry isolates, while human isolates showed the least genetic diversity. Cluster analysis obtained from RAPD_PCR and based on the genetic distances split the 49 strains into five distinct major clusters (A, B, C, D, and E). Cluster A and C were composed of isolates from poultry meat, cluster B was composed of isolates from human feces, cluster D was composed of isolates from minced meat, poultry meat and human feces and cluster E was composed of isolates from minced meat. Further characterization of these strains by using (GTG) 5 fingerprint repetitive sequence-based PCR analysis did not show further differentiation between various types of strains. To our knowledge, this is the first study in which the genetic diversity of C. perfringens isolates from different types of meats and human feces has been investigated.

Keywords: C. perfringens, genetic diversity, RAPD-PCR, REP-PCR

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Authors: M. G. Shehata, S. A. El Sohaimy, Marwa M. Abu-Serie, Nourhan M. Abd El-Aziz

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Probiotic strains can potentially be used as bio-preservatives and functional food supplement. Eight lactic acid bacteria strains (LAB) Lactobacillus brevis NRRL B-4527; Streptococcus thermophilus BLM 58; Pediococcusacidilactici ATCC 8042; Lactobacillus rhamnosus CCUG 1452; Lactobacillus curvatus ATCC 51436; Lactococcuslactis sub sp. lactisDSM 20481; Lactobacillus plantarum DMSZ 20079 and Lactobacillus plantarumTF103 were selected to screen the antioxidant, anticancer potential and probiotic properties. LAB strains exhibited good probiotic, antioxidant properties and showed antagonistic activity against food-borne pathogenic (Bacillus subtilis DB 100 host; Candida albicans ATCCMYA-2876; Clostridium botulinum ATCC 3584; Escherichia coli BA 12296; Klebsiellapneumoniae ATCC12296; Salmonella senftenberg ATCC 8400 and Staphylococcus aureus NCTC 10788). Further, in vitro probiotic properties of eight strains displayed excellent acid tolerance, bile tolerance, simulated gastrointestinal juice tolerance, in vitro adhesion ability for HT-29 cell line. The antioxidant effect of intracellular and cell-free extract of lactic acid bacteria strains was evaluated by various antioxidant assays, namely, resistance to hydrogen peroxide, DPPH radical scavenging, ABTS radical scavenging, and hydroxyl radical scavenging (HRS). The results showed that intracellular and cell-free supernatant of S. Thermophilus BLM 58, L. lactissubsp.lactis DSM 20481, P. acidilactici ATCC 8042, L. brevis NRRL B-4527 strains possess excellent antioxidant capacity. The intracellular of S. Thermophilus BLM 58 and P. acidilactici ATCC 8042 also showed excellent anticancer activity against Caco-2, MCF-7, HepG-2, and PC-3. Antioxidative property of selected lactic acid bacteria strains would be useful in the functional food manufacturing industry. They could beneficially affect the consumer by providing dietary source of antioxidants.

Keywords: anticancer activity, antioxidant activity, functional food, lactic acid bacteria, probiotic

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