Search results for: halotolerant pathogenic bacteria

Authors: Amel Bouderhem, Aminata Ould El Hadj Khelil, Amina N. Djrarbaoui, Aroussi Aroussi

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The present work aims to find the influence of the nature of the soil on the effectiveness of the biodegradation of hydrocarbons by a mixture of bacterial strains hydrocarbonoclastes. Processes of bioaugmentation and biostimulation trial are applied to samples of soils polluted voluntarily by the crude oil. For the evaluation of the biodegradation of hydrocarbons, the bacterial load, the pH and organic carbon total are followed in the different experimental batches. He bacterial load of the sandy soil varies among the witnesses of 45,2 .108 CFU/ml at the beginning of the experimentation to 214,07.108 CFU/ml at the end of the experiment. Of the soil silty-clay varies between 103,31 .108 CFU/ml and 614,86.108 CFU/ml . It was found a strong increase in the bacterial biomass during the processing of all samples. This increase is more important in the samples of sand bioaugmente or biomass increased from 63.16 .108 CFU/ml to 309.68 .108 CFU/ml than in soil samples silty clay- bioaugmente whose content in bacteria evolved of 73,01 .108 CFU/ml to 631.80 . 108CFU/ml

Keywords: pollution, hydrocarbons, bioremediation, bacteria hydrocarbonoclastes, ground, texture

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Authors: P. S. BEDI* , Neavty Thakur, Balvınder Sıngh

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In the present study an attempt has been made to prepare the crude extracts of leaves and stem of ‘Girardinia heterophylla’ by using various solvents like petroleum ether, ethanol and double distilled water. The samples were given the code NGLS 1, NGLS 2, NGLS 3, NGSS 1, NGSS 2 and NGSS 3 respectively. All the extracts were used to study their antimicrobial activity against gram positive bacteria eg. Bacillus subtilis, Gram negative bacteria eg. E. coli, K. pneumonia and antifungal activity against Aspergillus niger. The results of the antimicrobial activity showed that all the crude extracts of the plant posseses antibacterial activity. Maximum antibacterial activity was shown by NGLS 2, NGLS 3 and NGSS 3 against K. pneumonia. The growth of fungus A. niger was also inhibited by all the crude extracts. Maximum inhibition was shown by NGSS 2 followed by NGSS 1.

Keywords: Girardinia heterophylla, leaves and stem extracts, Antibacterial activity, antifungal activity.

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Authors: Iddrisu Ibrahim, Joseph Atia Ayariga, Junhuan Xu, Daniel Abugri, Boakai Robertson, Olufemi S. Ajayi

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The emergence of multidrug resistance poses a huge risk to public health globally. Yet these recalcitrant pathogens continue to rise in incidence rate, with resistance rates significantly outpacing the speed of antibiotic development. This, therefore, presents an aura of related health issues such as untreatable nosocomial infections arising from organ transplants and surgeries, as well as community-acquired infections that are related to people with compromised immunity, e.g., diabetic and HIV patients, etc. There is a global effort to fight multidrug-resistant pathogens spearheaded by the World Health Organization, thus calling for research into novel antimicrobial agents to fight multiple drug resistance. Previously, our laboratory demonstrated that Cannabidiol (CBD) was an effective antimicrobial against Salmonella typhimurium (S. typhimurium). However, we observed resistance development over time. To understand the mechanisms S. typhimurium uses to develop resistance to Cannabidiol (CBD), we studied the abundance of bacteria lipopolysaccharide (LPS) and membrane sterols of both susceptible and resistant S. typhimurium. Using real-time quantitative polymerase chain reaction (RT-qPCR), we also analyzed the expression of selected genes known for aiding resistance development in S. typhimurium. We discovered that there was a significantly higher expression of blaTEM, fimA, fimZ, and integrons in the CBD-resistant bacteria, and these were also accompanied by a shift in abundance in cell surface molecules such as lipopolysaccharide (LPS) and sterols.

Keywords: antimicrobials, resistance, cannabidiol, gram-negative bacteria, integrons, blaTEM, Fim, LPS, ergosterols

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Authors: Kanika Chowdhary, Satyawati Sharma

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Endophytic microbes are hosted inside plants in a symbiotic and hugely benefitting relationship. Exploring agriculturally beneficial endophytes is quite a prospective field of research. In the present work fungal endophytes associated with aromatic plant Ocimum basicilium L. were investigated for biocontrol potential. The anti-plant pathogenic activity of fungal endophytes was tested against causal agent of stem rot Sclerotinia sclerotiorum. 75 endophytic fungi were recovered through culture-dependent approach. Fungal identification was performed both microscopically and by rDNA ITS sequencing. Curvuaria lunata (Sb-6) and Colletotrichum lindemuthianum (Sb-8) inhibited 86% and 72% mycelia growth of S. sclerotinia on Sabouraud dextrose agar medium at 7.4 pH. Small-scale fermentation was carried out on sterilised oatmeal grain medium. In another set of experiment, fungi were grown in oatmeal grain medium amended with certain biostimulants such as aqueous seaweed extract (10% v/w); methanolic seaweed extract (5% v/w); cow urine (20% v/w); biochar (10% w/w) in triplicate along with control of each to ascertain the degree of metabolic difference and anti-plant pathogenic activity induced. Phytochemically extracts of both the fungal isolates showed the presence of flavanoids, phenols, tannins, alkaloids and terpenoids. Ethylacetate extract of C. lunata and C. lindemuthianum suppressed S. sclerotinia conidial germination at IC50 values of 0.514± 0.02 and 0.913± 0.04 mg/ml. Therefore, fungal endophytes of O. basicilium are highly promising bio-resource agent, which can be developed further for sustainable agriculture.

Keywords: endophytic fungi, ocimum basicilium, sclerotinia sclerotiorum, biostimulants

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Authors: Raana Babadi Fathipour

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Biofilms, those intricate structures of microbial aggregation that emerge as microorganisms adhere to animate or inanimate surfaces, possess an innate capacity to shield their inhabitants from adversities within the environment whilst fortifying their endurance against antimicrobial agents. This remarkable aspect facilitates the persistence and virulence of said microorganisms, establishing biofilm formation as an integral component of bacterial survival mechanisms. However, should foodborne pathogens adopt this mode of existence, the potentiality for foodborne disease infections becomes alarmingly intensified—an alarming prospect that harbors significant public health hazards and engenders deleterious economic ramifications. Thus, due to these consequences lurking on the horizon, extensive research concentrating upon comprehending biofilms and devising efficacious removal strategies assumes a position imbued with paramount importance within the realm of the food industry. The problem of food waste resulting from spoilage in the food industry continues to present a widespread challenge to both environmental sustainability and the security of our food supplies. In this comprehensive analysis, we delve into the formation of bacterial biofilms, highlighting the specific issues they pose within the realm of food production. Additionally, we provide an overview of various types of common foodborne pathogens that tend to thrive in these biofilms. Furthermore, we summarize existing strategies aimed at tackling or managing detrimental bacterial biofilm growth. We also introduce contemporary approaches that show promise in terms of controlling this issue and highlight their potential for further advancement. Ultimately, our focus lies on outlining prospects for future development as they pertain specifically to combatting bacterial biofilms within the field.

Keywords: foodborne pathogens, food safety, biofilm, resistance, quorum-sensing

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Authors: Sarah Devi Silvian, Hanna Shobrina Iqomatul Haq, Fara Cholidatun Nabila, Agustin Krisna Wardani

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Bacteria ability to survive in antibiotic is controlled by the presence of gene that encodes the antibiotic resistance protein. The instability of the antibiotic resistance gene can be observed by exposing the bacteria under the lethal dose of antibiotic. Low concentration of antibiotic can induce mutation, which may take a role in bacterial adaptation through the antibiotic concentration. Lactobacillus casei FNCC 0090 is one of the probiotic bacteria that has an ability to survive in tetracycline by expressing the tetM gene. The aims of this study are to observe the possibilities of mutation happened in L.casei FNCC 0090 by exposing in sub-lethal dose of tetracycline and also observing the instability of the tetM gene by comparing the sequence between the wild type and mutant. L.casei FNCC 0090 has a lethal dose in 60 µg/ml, low concentration is applied to induce the mutation, the range from 10 µg/ml, 15 µg/ml, 30 µg/ml, 45 µg/ml, and 50 µg/ml. L.casei FNCC 0090 is exposed to the low concentration from lowest to the highest concentration to induce the adaptation. Plasmid is isolated from the highest concentration culture which is 50 µg/ml by using modified alkali lysis method with the addition of lysozyme. The tetM gene is isolated by using PCR (Polymerase Chain Reaction) method, then PCR amplicon is purified and sequenced. Sequencing is done on both samples, wild type and mutant. Both sequences are compared and the mutations can be traced in the presence of nucleotides changes. The changing of the nucleotides means that the tetM gene is instable.

Keywords: L. casei FNCC 0090, probiotic, tetM, tetracycline

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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: Mohammad Hossein Pazandeh, Monir Doudi, Sona Rostampour Yasouri

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Irregular consumption of current antibiotic makes increases of antibiotic resistance between urin pathogens on all worlds. This study selected based on this great community problem. The aim of this study was the biosynthesis of silver nanoparticles from Zataria multiflora extract and then to investigate its antibacterial effect on gram-negative bacilli common in Urinary Tract Infections (UTI) and MDR. The plant used in the present research was Zataria multiflora whose extract was prepared through Soxhlet extraction method. Green synthesis condition of silver nanoparticles was investigated in terms of three parameters including the extract amount, concentration of silver nitrate salt, and temperature. The seizes of nanoparticles were determined by Zetasizer. In order to identify synthesized silver nanoparticles Transmission Electron Microscopy (TEM) and X-ray Diffraction (XRD) methods were used. For evaluating the antibacterial effects of nanoparticles synthesized through biological method different concentrations of silver nanoparticles were studied on 140 cases of Muliple Drug Resistance (MDR) bacteria strains Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, Proteus vulgaris,Citrobacter freundii, Acinetobacter bumanii and Pseudomonas aeruginosa, (each genus of bacteria, 20 samples), which all were MDR and cause urinary tract infections , for identification of bacteria were used of Polymerase Chain Reaction (PCR) test and laboratory methods (Agar well diffusion and Microdilution methods) to assess their sensitivity to Nanoparticles. The data were analyzed using SPSS software by nonparametric Kruskal-Wallis and Mann-Whitney tests. Significant results were found about the effects of silver nitrate concentration, different amounts of Zataria multiflora extract, and temperature on nanoparticles; that is, by increasing the concentration of silver nitrate, extract amount, and temperature, the sizes of synthesized nanoparticles declined. However, the effect of above mentioned factors on particles diffusion index was not significant. Based on the TEM results, particles were mainly spherical shape with a diameter range of 25 to 50 nm. The results of XRD Analysis indicated the formation of Nanostructures and Nanocrystals of silver.. The obtained results of antibacterial effects of different concentrations of silver nanoparticles on according to agar well diffusion and microdilution method, biologically synthesized nanoparticles showed 1000 mg /ml highest and lowest mean inhibition zone diameter in E.coli , Acinetobacter bumanii 23 and 15mm, respectively. MIC was observed for all of bacteria 125mg/ml and for Acinetobacter bumanii 250mg/ml.Comparing the growth inhibitory effect of chemically synthesized Nanoparticles and biologically synthesized Nanoparticles showed that in the chemical method the highest growth inhibition belonged to the concentration of 62.5 mg /ml. The inhibitory effect on the growth all of bacteria causes of urine infection and MDR was observed and by increasing silver ion concentration in Nanoparticles, antibacterial activity increased. Generally, the biological synthesis can be considered an efficient way not only in making Nanoparticles but also for having anti-bacterial properties. It is more biocompatible and may be possess less toxicity than the Nanoparticles synthesized chemically.

Keywords: biosynthesis, MDR bacteria, silver nanoparticles, UTI

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Authors: Zuliani Mahmood, Thirumulu Ponnuraj Kannan, Yean Yean Chan, Salahddin A. Al-Hudhairy

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Caries is the most common childhood disease which develops due to disturbances in the physiological equilibrium in the dental plaque resulting in demineralization of tooth structures. Plaque and dentine samples were collected from three different tooth surfaces representing caries progression (intact, over carious lesion and dentine) in children with early childhood caries (ECC, n=36). In caries free (CF) children, plaque samples were collected from sound tooth surfaces at baseline and after one year (n=12). The genomic DNA was extracted from all samples and subjected to 16S rRNA PCR amplification. The end products were cloned into pCR®2.1-TOPO® Vector. Five randomly selected positive clones collected from each surface were sent for sequencing. Identification of the bacterial clones was performed using BLAST against GenBank database. In the ECC group, the frequency of Lactobacillus sp. detected was significantly higher in the dentine surface (p = 0.031) than over the cavitated lesion. The highest frequency of bacteria detected in the intact surfaces was Fusobacterium nucleatum subsp. polymorphum (33.3%) while Streptococcus mutans was detected over the carious lesions and dentine surfaces at a frequency of 33.3% and 52.7% respectively. Fusobacterium nucleatum subsp. polymorphum was also found to be highest in the CF group (41.6%). Follow up at the end of one year showed that the frequency of Corynebacterium matruchotii detected was highest in those who remained caries free (16.6%), while Porphyromonas catoniae was highest in those who developed caries (25%). In conclusion, Streptococcus mutans and Porphyromonas catoniae are strongly associated with caries progression, while Lactobacillus sp. is restricted to deep carious lesions. Fusobacterium nucleatum subsp. polymorphum and Corynebacterium matruchotii may play a role in sustaining the healthy equilibrium in the dental plaque. These identified bacteria show promise as potential biomarkers in diagnosis which could help in the management of dental caries in children.

Keywords: early childhood caries, genotypic identification, oral bacteria, 16S rRNA

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Authors: Wendu Admasu, Assefa Sintayehu, Alemu Gezahgne, Zewdu Terefework

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Eucalyptus is the most widely planted forest tree species in the world. In Ethiopia, pathogenic fungi pose an increasing threat to Eucalyptus species. Due to limited research, there is insufficient information on the associated diseases and pathogens. This study investigated Eucalyptus diseases, the extent of their damage, and the causal fungal pathogens. A Eucalyptus disease survey was conducted in the Eucalyptus forestry areas of Ethiopia during the growth years 2019/20 and 2020/21. Disease assessment and sampling were carried out in eighteen plantations at nine locations. E. camaldulensis was the most dominant species planted in the surveyed areas. The field study shows a high incidence and severity of canker diseases. Diseased stem and branch samples were collected, cultured on malt extract agar media and studied. The results of morphological and ITS sequence analysis confirmed that the fungal species Neofusicoccum parvum, Lasiodiplodia theobromae, and Aplosporella hesperidica caused the observed canker symptoms. This is the first report of Lasiodiplodia theobromae and Aplosporella hesperidica causing diseases in Eucalyptus plants in Ethiopia. Changes in global climate and environmental factors, such as altitude, are believed to have a strong impact on the susceptibility of Eucalyptus plants to diseases. Strict quarantine practices and continuous monitoring of pathogenic and endophytic fungal species associated with Eucalyptus trees are issued to be prioritized to effectively control and manage the disease.

Keywords: Neofusicoccum, Lasiodiplodia, Aplosporella, pathogenicity, phylogeny, severity

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Authors: Wahiba Boudraa, Farah Chettibbi, Meriem Aberkane, Fatma Djamaa, Moussa Houhamdi

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The intensity of human activities in regions surrounding the Mediterranean Sea always has a strong long-term environmental impact resulting in coastal and marine degradation, as well as an aggravated risk of more serious damage. The available data on water quality show that most water resources in Algeria are polluted by uncontrolled discharges from municipal sewage and untreated industrial effluents. Annaba is a coastal town in Algeria; The Gulf of Annaba, responds to these changes as it receives the continental inputs and urban waste, industrial without prior treatment of a highly industrialized and urbanized city, subject to the same environmental problems that know the rest of the Algerian coast. In later year, the beaches of bacterial enumeration process waters showed relatively high levels of bacterial indicators of fecal contamination (group D streptococci, total and fecal coliforms), which reflect the risks to people attending these beaches. During the twelve months of our study, we isolated from three beaches in the city of Annaba (St. Cloud, El-Kettara, and Djenane El Bey) a number of pathogenic microorganisms considered, namely: Salmonella, Aeromonas, Citrobacter, Yersinia, Enterococcus, and E.coli. The microbial count revealed elevated levels of coliform bacteria, fecal coliforms and fecal streptococci quite high especially in urban beaches (St. Cloud and El-Kettara). They are widely popular during the summer by many vacationers. For the physico-chemical parameters, there exist some weak values which increase during the pluvial period, hivernal and festival saison. These values remain, nevertheless, weak to be able to cause an organic or metallic pollution.

Keywords: quality microbiology, pollution of water, fecal contamination, physico-chemistry, beaches of Annaba city, Algeria.

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Authors: Qunying Yuan, Manjula Bomma, Adrian Rhoden, Zhigang Xiao

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Selenium is an essential micronutrient for all mammals and plays an important role in maintaining human physiological functions. The potential applications of selenium as food supplements, cancer-prevention, antimicrobial and anti-inflammatory agents have been investigated in biomedicine and food sciences. Nanoscale of selenium is of particular interest due to its better biocompatibility, higher bioavailability, lower toxicity, more homogeneous distribution, and presumptive controlled release of substances. The objective of this study is to explore whether selenium nanoparticle (SeNP) has the potential to be used as a food preservative to reduce food spoilage. SeNPs were synthesized through ascorbic acid reduction of sodium selenite using the bovine serum albumin (BSA) as capping and stabilizing agent. The chemically synthesized SeNPs had a spherical conformation and a size of 22.8 ± 4.7 nm. FTIR analysis confirmed that the nanoparticles were covered with BSA. We further tested the antimicrobial activity of these SeNPs against common food-borne bacteria. Colony forming unit assay showed that SeNPs exhibited good inhibition on the growth of Listeria Monocytogens (ATCC15313), Staphylococcus epidermidis (ATCC 700583) starting at 0.5µg/mL, but only a moderate inhibitory effect on the growth of Staphylococcus aureus (ATCC12600) and Vibrio alginolyticus (ATCC 33787) at a concentration higher than 10µg/mL and 2.5µg/mL, respectively. There was a mild effect against the growth Salmonella enterica (ATCC19585) when the concentration reached 15µg/mL. No inhibition was observed in the growth of Enterococcus faecalis (ATCC 19433). Surprisingly, SeNPs appeared to promote the growth of Vibrio parahaemolyticus (ATCC43996) and Salmonella enterica (ATCC49284) at 30 µg/mL and above. Our preliminary data suggested that the chemically synthesized SeNPs may be able to inhibit some food-borne bacteria, and SeNP as a food preservative should be used with caution. We will explore the mechanisms of the inhibitory action of chemically synthesized SeNPs on bacterial growth and whether the SeNPs are able to inhibit the development of biofilm and antibiotic resistance.

Keywords: antimicrobial, food-borne bacteria, nanoparticles, selenium

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Authors: Sonya Barzgar, J. Patrick, A. Hettiaratchi

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Methane (CH4) is reputed as the second largest contributor to greenhouse effect with a global warming potential (GWP) of 34 related to carbon dioxide (CO2) over the 100-year horizon, so there is a growing interest in reducing the emissions of this gas. Methane biofiltration (MBF) is a cost effective technology for reducing low volume point source emissions of methane. In this technique, microbial oxidation of methane is carried out by methane-oxidizing bacteria (methanotrophs) which use methane as carbon and energy source. MBF uses a granular medium, such as soil or compost, to support the growth of methanotrophic bacteria responsible for converting methane to carbon dioxide (CO₂) and water (H₂O). Even though the biofiltration technique has been shown to be an efficient, practical and viable technology, the design and operational parameters, as well as the relevant microbial processes have not been investigated in depth. In particular, limited research has been done on the effects of sulphur on methane bio-oxidation. Since bacteria require a variety of nutrients for growth, to improve the performance of methane biofiltration, it is important to establish the input quantities of nutrients to be provided to the biofilter to ensure that nutrients are available to sustain the process. The study described in this paper was conducted with the aim of determining the influence of sulphur on methane elimination in a biofilter. In this study, a set of experimental measurements has been carried out to explore how the conversion of elemental sulphur could affect methane oxidation in terms of methanotrophs growth and system pH. Batch experiments with different concentrations of sulphur were performed while keeping the other parameters i.e. moisture content, methane concentration, oxygen level and also compost at their optimum level. The study revealed the tolerable limit of sulphur without any interference to the methane oxidation as well as the particular sulphur concentration leading to the greatest methane elimination capacity. Due to the sulphur oxidation, pH varies in a transient way which affects the microbial growth behavior. All methanotrophs are incapable of growth at pH values below 5.0 and thus apparently are unable to oxidize methane. Herein, the certain pH for the optimal growth of methanotrophic bacteria is obtained. Finally, monitoring methane concentration over time in the presence of sulphur is also presented for laboratory scale biofilters.

Keywords: global warming, methane biofiltration (MBF), methane oxidation, methanotrophs, pH, sulphur

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Authors: Eszter J. Tóth, Alexandra Hoffmann, Csaba Vágvölgyi, Tamás Papp

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Members of the genera Curvularia and Bipolaris are closely related melanin producing filamentous fungi; both of them have the teleomorph states in genus Cochliobolus. While Bipolaris species infect only plants and may cause serious agriculture damages, some Curvularia species was recovered from opportunistic human infections. The human pathogenic species typically cause phaeohyphomycoses, i.e. mould infections caused by melanised fungi, which can manifest as invasive mycoses with frequent involvement of the central nervous system in immunocompromised patients or as local infections (e.g. keratitis, sinusitis, and cutaneous lesions) in immunocompetent people. Although their plant-fungal interactions have been intensively studied, there is only little information available about the human pathogenic feature of these fungi. The aim of this study was to investigate the neutrophil granulocytes’ response to hyphal forms of Curvularia and Bipolaris in comparison with the response to Aspergillus. In the present study Curvularia lunata SZMC 23759 and Aspergillus fumigatus SZMC 23245 both isolated from human eye infection, and Bipolaris zeicola BRIP 19582b isolated from plant leaf were examined. Neutrophils were isolated from heparinised venous blood of healthy donors with dextran sedimentation followed by centrifugation over Ficoll and hypotonic lysis of erythrocytes. Viability and purity of the cells were checked with trypan blue and Wright staining, respectively. Infection of neutrophils was carried out with germinated conidia in a ratio of 5:1. Production of hydrogen peroxide, superoxide anion, and nitrogen monoxide was measured both intracellularly and extracellularly in response to the germinated spores with or without the supernatant and after serum treatment. ROS and NOS production of neutrophils in interaction with the three fungi were compared. It is already known that Aspergillus species induce ROS production of neutrophils only after serum treatment. Although, in case of C. lunata, serum opsonisation also induced an intensive production of reactive species, lower level of production was measured in the lack of serum as well. After interaction with the plant pathogenic B. zeicola, amount of reactive species found to be similar with and without serum treatment. The presence of germination supernatant decreased the reactive species production in case of each fungus. Interaction with Curvularia, Bipolaris and Aspergillus species induced different response of neutrophils. It seems that recognition of C. lunata and B. zeicola is independent of serum opsonisation, albeit it increases the level of the produced reactive species in response for C. lunata. The study was supported by the grant LP2016-8/2016.

Keywords: Curvularia, neutrophils, NOS, ROS, serum opsonisation

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Authors: O. Zennouhi, M. El Mderssa, J. Ibijbijen, E. Bouiamrine, L. Nassiri

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Rangelands and silvopastoral systems of the middle Atlas are under a heavy pressure, which led to pasture degradation, invasion by non-palatable and toxic species and edaphic aridification due to the regression of the global vegetation cover. In this situation, the introduction of multipurpose leguminous shrubs, such as Bituminaria bituminosa (L.) Stirton, commonly known as bituminous clover, could be a promising socio-ecological alternative for the rehabilitation of these degraded areas. The application of biofertilizers like plant growth promoting rhizobacteria especially phosphate solubilizing bacteria (PSB) can ensure a successful installation of this plant in the selected degraded areas. The main objective of the present work is to produce well-inoculated seedlings using the best efficient PSB strains in the greenhouse to increase their ability to resist to environmental constraints once transplanted to the field in the central Middle Atlas.

Keywords: biofertilizers, bituminaria bituminosa, phosphate solubilizing bacteria, rehabilitation

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Authors: Tomasz Borowski, Dawid Sołoducha, Agata Markowska-Szczupak, Aneta Wesołowska, Marian Kordas, Rafał Rakoczy

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Essential oils (EOs) are fragrant volatile oils obtained from plants. These are used for cooking (for flavor and aroma), cleaning, beauty (e.g., rosemary essential oil is used to promote hair growth), health (e.g. thyme essential oil cures arthritis, normalizes blood pressure, reduces stress on the heart, cures chest infection and cough) and in the food industry as preservatives and antioxidants. Rosemary and thyme essential oils are considered the most eminent herbs based on their history and medicinal properties. They possess a wide range of activity against different types of bacteria and fungi compared with the other oils in both in vitro and in vivo studies. However, traditional uses of EOs are limited due to rosemary and thyme oils in high concentrations can be toxic. In light of the accessible data, the following hypothesis was put forward: Low frequency rotating magnetic field (RMF) increases the antimicrobial potential of EOs. The aim of this work was to investigate the antimicrobial activity of commercial Salvia Rosmarinus L. and Thymus vulgaris L. essential oil from Polish company Avicenna-Oil under Rotating Magnetic Field (RMF) at f = 25 Hz. The self-constructed reactor (MAP) was applied for this study. The chemical composition of oils was determined by gas chromatography coupled with mass spectrometry (GC-MS). Model bacteria Escherichia coli K12 (ATCC 25922) was used. Minimum inhibitory concentrations (MIC) against E. coli were determined for the essential oils. Tested oils in very small concentrations were prepared (from 1 to 3 drops of essential oils per 3 mL working suspensions). From the results of disc diffusion assay and MIC tests, it can be concluded that thyme oil had the highest antibacterial activity against E. coli. Moreover, the study indicates the exposition to the RMF, as compared to the unexposed controls causing an increase in the efficacy of antibacterial properties of tested oils. The extended radiation exposure to RMF at the frequency f= 25 Hz beyond 160 minutes resulted in a significant increase in antibacterial potential against E. coli. Bacteria were killed within 40 minutes in thyme oil in lower tested concentration (1 drop of essential oils per 3 mL working suspension). Rapid decrease (>3 log) of bacteria number was observed with rosemary oil within 100 minutes (in concentration 3 drops of essential oils per 3 mL working suspension). Thus, a method for improving the antimicrobial performance of essential oil in low concentrations was developed. However, it still remains to be investigated how bacteria get killed by the EOs treated by an electromagnetic field. The possible mechanisms relies on alteration in the permeability of ionic channels in ionic channels in the bacterial cell walls that transport in the cells was proposed. For further studies, it is proposed to examine other types of essential oils and other antibiotic-resistant bacteria (ARB), which are causing a serious concern throughout the world.

Keywords: rotating magnetic field, rosemary, thyme, essential oils, Escherichia coli

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Authors: Zakaria Boual, Abdellah Kemassi, Toufik Chouana, Philippe Michaud, Mohammed Didi Ould El Hadj

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In order to investigate the prebiotic potential of oligosaccharides prepared by chemical hydrolysis of water-soluble polysaccharides (WSP) from Zizyphus lotus leaves, the effect of oligosaccharides on bacterial growth was studied. The chemical composition of WSP was evaluated by colorimetric assays revealed the average values: 7.05±0.73% proteins and 86.21±0.74% carbohydrates, among them 64.81±0.42% are neutral sugar and the rest 16.25±1.62% are uronic acids. The characterization of monosaccharides was determined by high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) was found to be composed of galactose (23.95%), glucose (21.30%), rhamnose (20.28%), arabinose (9.55%), and glucuronic acid (22.95%). The effects of oligosaccharides on the growth of lactic acid bacteria were compared with those of fructo-oligosaccharide (RP95). The oligosaccharides concentration was 1g/L of man rogosa sharpe broth. Bacterial growth was assessed during 2, 4.5, 6.5, 9, 12, 16 and 24 h by measuring the optical density of the cultures at 600 nm (OD600) and pH values. During fermentation, pH in broth cultures decreased from 6.7 to 5.87±0.15. The enumeration of lactic acid bacteria indicated that oligosaccharides led to a significant increase in bacteria (P≤0.05) compared to the control. The fermentative metabolism appeared to be faster on RP95 than on oligosaccharides from Zizyphus lotus leaves. Both RP95 and oligosaccharides showed clear prebiotic effects, but had differences in fermentation kinetics because of to the different degree of polymerization. This study shows the prebiotic effectiveness of oligosaccharides, and provides proof for the selection of leaves of Zizyphus lotus for use as functional food ingredients.

Keywords: Zizyphus lotus, polysaccharides, characterization, prebiotic effects

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Authors: Afzan Mahmad, Santibuana Abd Rahman, Gouri Kumar Dash, Mohd. Syafiq Bin Abdullah

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Antibacterial activity of the methanol extract of fruit peel of Punica granatum Linn (Family: Punicaceae) was evaluated against two Gram positive and two Gram negative bacteria. The Gram positive bacteria included Staphylococcus aureus, Streptococcus pneumoniae and the Gram negative organisms included Escherichia coli and Pseudomonas aeruginosa respectively. The culture media used for antibacterial assay was Mueller Hinton agar for the growth of S. aureus, E. coli, and P. aeruginosa. The media used for the growth of S. pneumoniae was Mueller Hinton blood agar. The antibacterial assay was performed through Disc diffusion technique. The methanol extract was tested at three different concentrations (50, 100 and 200 mg/ml). Standard antibiotic discs containing vancomycin (30 μg) for S. pneumoniae, penicillin (10 units) for S. aureus, ceftriaxone (30 μg) for E. coli and ciprofloxacin (5 μg) for P. aeruginosa were used for the activity comparison. The results of the study revealed that the extract possesses antibacterial activity against S. aureus, S. pneumoniae and P. aeruginosa at all tested concentrations. The maximum zone of inhibition of 19 mm of the extract at 200 mg/ml was observed against S. pneumoniae. However, no zone of inhibition was observed against E. coli at the tested concentrations of the extract. Based on the results obtained in this study, it may be concluded that the fruit peel of P. granatum possess broad spectrum of antibacterial activity against a number bacteria.

Keywords: Punica granatum Linn., methanol extract, antibacterial, zone of inhibition

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Authors: Vitheejongjaroen P., Jaisin Y., Pachekrepapol U., Taweechotipatr M.

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Lactic acid bacteria (LAB) are the most common type of microorganisms that had been used as probiotics also known for many beneficial health effects. The antioxidant activity of LAB is associated with numerous health-protective effects. This research aimed to investigate the antioxidant activity of lactic acid bacteria isolated from Thai sour pork sausage for their application in fermented milk products. Antioxidant activity determined by DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay showed that the isolate FN33-7, as 1 of 8 isolated exhibited scavenging activity in intact cell 5-7%, and supernatant 13-16%, intracellular cell free extract 42-48% respectively. This isolate was identified using 16S ribosomal DNA sequence analysis as Lactobacillus plantarum. The effect of milk fermented with L. plantarum FN33-7 on microbial count, pH and syneresis was assessed during refrigerated storage period of 28 days. The strain showed increased viability, pH level decreased, while syneresis increased. These results are similar to dairy products fermented with commercial starter cultures. Additionally, microstructure analysis of fermented milk by fluorescent microscopy showed that curd structure appeared to be dense and less porous in this fermented milk than commercial yogurt. The results of this study indicated that L. plantarum FN33-7 was a good probiotic candidate to be used in cultured milk products to reduce the risk of diseases caused by oxidative stress.

Keywords: Lactobacillus plantarum, probiotics, free radical, antioxidant, oxidative stress, fermented milk products

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Authors: Maha Alaoui Ismaili, Bouchta Saidi, Mohamed Zahar, Abed Hamama

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112 lactic isolates were obtained from 15 samples of camel raw milk produced in Laayoune Boujdour Sakia-El Hamra region (South of Morocco). The main objective was the identification of species of lactic flora belonging to Lactococcus, Lactobacillus and Leuconostoc. Data obtained showed predominance of cocci among lactic isolates (86.6%) while lactic rods represented only 13.4%. With regard to genera identified, Enterococcus was the mostly found out (53.57%), followed by Lactococcus (28.57%), Lactobacillus (13.4%) and Leuconostoc (4.4 %). Identification of the lactic isolates according to their morphological, physiological, and biochemical characteristics led to differentiating 11 species with Lactococcus lactis ssp lactis biovar diacetylactis being the mostly encountered (24.1%) followed by Lactobacillus brevis (3.57%), Lactobacillus plantarum (3.57%), Lactobacillus delbrueckii subsp lactis (3.57%) and Lactococcus lactis subsp cremoris (2.67%).

Keywords: raw camel milk, south of morocco, lactic acid bacteria, identification

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Authors: Ilze Dimanta, Zane Rutkovska, Vizma Nikolajeva, Janis Kleperis, Indrikis Muiznieks

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Majority of word’s steadily increasing energy consumption is provided by non-renewable fossil resources. Need to find an alternative energy resource is essential for further socio-economic development. Hydrogen is renewable, clean energy carrier with high energy density (142 MJ/kg, accordingly – oil has 42 MJ/kg). Biological hydrogen production is an alternative way to produce hydrogen from renewable resources, e.g. using organic waste material resource fermentation that facilitate recycling of sewage and are environmentally benign. Hydrogen gas is produced during the fermentation process of bacteria in anaerobic conditions. Bacteria are producing hydrogen in the liquid phase and when thermodynamic equilibrium is reached, hydrogen is diffusing from liquid to gaseous phase. Because of large quantities of available crude glycerol and the highly reduced nature of carbon in glycerol per se, microbial conversion of it seems to be economically and environmentally viable possibility. Such industrial organic waste product as crude glycerol is perspective for usage in feedstock for hydrogen producing bacteria. The process of biodiesel production results in 41% (w/w) of crude glycerol. The developed lab-scale test system (experimental bioreactor) with hydrogen micro-electrode (Unisense, Denmark) was used to determine hydrogen production yield and rate in the liquid phase. For hydrogen analysis in the gas phase the RGAPro-100 mass-spectrometer connected to the experimental test-system was used. Fermentative bacteria strains were tested for hydrogen gas production rates. The presence of hydrogen in gaseous phase was measured using mass spectrometer but registered concentrations were comparatively small. To decrease the hydrogen partial pressure in liquid phase reactor with a system for continuous bubbling with inert gas was developed. H2 production rate for the best producer in liquid phase reached 0,40 mmol H2/l, in gaseous phase - 1,32 mmol H2/l. Hydrogen production rate is time dependent – higher rate of hydrogen production is at the fermentation process beginning when concentration increases, but after three hours of fermentation, it decreases.

Keywords: bio-hydrogen, fermentation, experimental bioreactor, crude glycerol

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Authors: Nurcan Cetinkaya

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The aim of this paper is to bring about the utilization of Juncus acutus as an alternative roughage resource in ruminant nutrition. In Turkey, JA is prevailing plant of the natural grassland in Kizilirmak Delta, Samsun. Crude nutrient values such as crude protein (CP), ether extract (EE), organic matter (OM), neutral detergent fiber (NDF), acid detergent fiber (ADF), and acid detergent lignin(ADL) including antioxidant activity, total phenolic and flavonoid compounds, total organic matter digestibility (OMD) and metabolisable energy (ME) values of Juncus acutus stem, seed, and also its mixture with maize silage were estimated. and published. Furthermore, the effects of JA over rumen cellulolitic bacteria were studied. The obtained results from different studies conducted on JA by our team show that Juncus acutus may be a new roughage source in ruminant nutrition.

Keywords: antioxidant activity, cellulolytic bacteria, Juncus acutus, organic matter digestibility

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Authors: Ágota Ábrahám, Md Nurul Islam, Karimane Zeghbib, Gábor Kemenesi, Sazeda Akter

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Collecting palm sap as a food source is an everyday practice in some parts of the world. However, the consumption of palm juice has been associated with regular infections and epidemics in parts of Bangladesh. This is attributed to fruit-eating bats and other vertebrates or invertebrates native to the area, contaminating the food with their body secretions during the collection process. The frequent intake of palm juice, whether as a processed food product or in its unprocessed form, is a common phenomenon in large areas. The range of pathogens suitable for human infection resulting from this practice is not yet fully understood. Additionally, the high sugar content of the liquid makes it an ideal culture medium for certain bacteria, which can easily propagate and potentially harm consumers. Rapid diagnostics, especially in remote locations, could mitigate health risks associated with palm juice consumption. The primary objective of this research is the rapid genomic detection and risk assessment of bacteria that may cause infections in humans through the consumption of palm juice. Utilizing state-of-the-art third-generation Nanopore metagenomic sequencing technology based on 16S rRNA, and identified bacteria primarily involved in fermenting processes. The swift metagenomic analysis, coupled with the widespread availability and portability of Nanopore products (including real-time analysis options), proves advantageous for detecting harmful pathogens in food sources without relying on extensive industry resources and testing.

Keywords: raw date palm sap, NGS, metabarcoding, food safety

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Authors: Mohamed Abd Elfattah Ibraheem Elghrbawy

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Biodeterioration of cultural heritage is a complex process which is caused by the interaction of many physical, chemical and biological agents; the growth of microorganisms can cause staining, cracking, powdering, disfigurement and displacement of monuments material, which leads to the permanent loss of monuments material. Organisms causing biodeterioration on monuments have usually been controlled by chemical products (biocides). In order to overcome the impact of biocides on the environment, human health and monument substrates, alternative tools such as antimicrobial agents from natural products can be used for monuments conservation and protection. The problem is how to formulate antibacterial agents with high efficiency and low toxicity. Various types of biodegradable metal nanoparticles (MNPs) have many applications in plant extract delivery. So, Nano-encapsulation of metal and natural antimicrobial agents using polymers such as chitosan increases their efficacy, specificity and targeting ability. Green synthesis and characterization of metal nanoparticles such as silver with natural products extracted from some plants having antimicrobial properties, using the ecofriendly method one pot synthesis. Encapsulation of the new synthesized mixture using some biopolymers such as chitosan nanoparticles. The dispersions and homogeneity of the antimicrobial heterogeneous metal nanoparticles encapsulated by biopolymers will be characterized and confirmed by Fourier Transform Infrared Spectroscopy (FTIR), Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM) and Zeta seizer. The effect of the antimicrobial biopolymer metal nano-formulations on normal human cell lines will be investigated to evaluate the environmental safety of these formulations. The antimicrobial toxic activity of the biopolymeric antimicrobial metal nanoparticles formulations will be will be investigated to evaluate their efficiency towards different pathogenic bacteria and fungi.

Keywords: antimicrobial, biodeterioration, chitosan, cultural heritage, silver

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Authors: Muhammad Shahzad Hussain

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Two major problems are facing generally by conventional poultry farming that is disease outbreaks and poor performance, which results due to improper management. To enhance the growth performance and efficiency of feed and reduce disease outbreaks, antibiotic growth promoters (AGPs) which are antibiotics at sub-therapeutic levels, are extensively used in the poultry industry. European Union has banned the use of antibiotics due to their presence in poultry products, development of antibiotic-resistant pathogens, and disturbance of normal gut microbial ecology. These residues cause serious health concerns and produce antibiotic resistance in pathogenic microbes in human beings. These issues strengthen the need for the withdrawal of AGPs from poultry feed. Nowadays, global warming is a major issue, and it is more critical in tropical areas like Pakistan, where heat stress is already a major problem. Heat stress leads to poor production performance, high mortality, immuno-suppression, and concomitant diseases outbreak. The poultry feed industry in Pakistan, like other countries of the world, has been facing shortages and high prices of local as well as imported feed ingredients. Prebiotics are potential replacer for AGP as prebiotics has properties to enhance the production potential and reduce the growth of harmful bacteria as well as stimulate the growth/activity of beneficial bacteria. The most commonly used prebiotics in poultry includes mannan oligosaccharide (MOS). MOS is an essential component of the yeast cell wall (YCW) (Saccharomyces cerevisiae); therefore, the YCW wall possesses prebiotic properties. The use of distillery yeast wall (YCW) has the potential to replace conventional AGPs and to reduce mortality due to heat stress as well as to bind toxins in the feed. The dietary addition of YCW has not only positive effects on production performance in poultry during normal conditions but during stressful conditions. A total of 168-day-old broilers were divided into 6 groups, each of which has 28 birds with 4 replicates (n=7).Yeast cell wall (YCW) supplementation @ 0%, 1%, 1.5%, 2%, 2.5%, 3% from day 0 to 35. Heat stress was exposed from day 21 to 35 at 30±1.1ᵒC with relative humidity 65±5%. Zootechnical parameters like body weight, FCR, Organ development, and histomorphometric parameters were studied. A significant weight gain was observed at group C supplemented @ 1.5% YCW during the fifth week. Significant organ weight gain of Gizzard, spleen, small intestine, and cecum was observed at group C supplemented @ 1.5% YCW. According to morphometric indices Duodenum, Jejunum, and Ileum has significant villus height, while Jejunum and Ileum have also significant villus surface area in the group supplemented with 1.5% YCW. IEL count was only decreased in 1.5% YCW-fed group in jejunum and ileum, not in duodenum, that was less in 2% YCW-supplemented group. Dietary yeast cell wall of saccharomyces cerevisiae partially reduced the effects of high ambient temperature in terms of better growth and modified gut histology and components of mucosal immune response to better withstand heat stress in broilers.

Keywords: antibiotics, AGPs, broilers, MOS, prebiotics, YCW

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Authors: Claudio Lamilla, Andrés Santos, Vicente Llanquinao, Jocelyn Hermosilla, Leticia Barrientos

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The industry in general is very interested in improving and optimizing industrial processes in order to reduce the costs involved in obtaining raw materials and production. Thus, an interesting and cost-effective alternative is the incorporation of bioactive metabolites in such processes, being an example of this enzymes which catalyze efficiently a large number of enzymatic reactions of industrial and biotechnological interest. In the search for new sources of these active metabolites, Antarctica is one of the least explored places on our planet where the most drastic cold conditions, salinity, UVA-UVB and liquid water available are present, features that have shaped all life in this very harsh environment, especially bacteria that live in different Antarctic ecosystems, which have had to develop different strategies to adapt to these conditions, producing unique biochemical strategies. In this work the production of cellulolytic enzymes of seven bacterial strains isolated from marine sediments at different sites in the Antarctic was evaluated. Isolation of the strains was performed using serial dilutions in the culture medium at M115°C. The identification of the strains was performed using universal primers (27F and 1492R). The enzyme activity assays were performed on R2A medium, carboxy methyl cellulose (CMC)was added as substrate. Degradation of the substrate was revealed by adding Lugol. The results show that four of the tested strains produce enzymes which degrade CMC substrate. The molecular identifications, showed that these bacteria belong to the genus Streptomyces and Pseudoalteromonas, being Streptomyces strain who showed the highest activity. Only some bacteria in marine sediments have the ability to produce these enzymes, perhaps due to their greater adaptability to degrade at temperatures bordering zero degrees Celsius, some algae that are abundant in this environment and have cellulose as the main structure. The discovery of new enzymes adapted to cold is of great industrial interest, especially for paper, textiles, detergents, biofuels, food and agriculture. These enzymes represent 8% of industrial demand worldwide and is expected to increase their demand in the coming years. Mainly in the paper and food industry are required in extraction processes starch, protein and juices, as well as the animal feed industry where treating vegetables and grains helps improve the nutritional value of the food, all this clearly puts Antarctic microorganisms and their enzymes specifically as a potential contribution to industry and the novel biotechnological applications.

Keywords: antarctic, bacteria, biotechnological, cellulolytic enzymes

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Authors: Andreev V. A., Kovalenko T. N., Privolnev V. V., Chernavin A. V., Knyazeva E. R.

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Aim: To evaluate the antagonistic activity of Streptococcus salivarius K12 (SsK12) against ENT and oral cavity infection pathogens (S. pneumoniae, S. pyogenes, S. aureus), gram-negative bacteria (E. coli, P. aeruginosa) and C. albicans. Materials and methods: The probiotic strain SsK12 was isolated from the dietary supplement containing at least 1 × 109 CFU per tablet. The tablet was dissolved in the enrichment broth. The resulting suspension was seeded on 5% blood agar and incubated at 35°C in 4-6% CO2 for 48 hours. The raised culture was identified as Streptococcus salivarius with MALDI-TOF mass spectrometry method. The evaluation of SsK12 antagonistic activity was carried out using a perpendicular streak technique. The daily SsK12 culture was inoculated as heavy streaks with a loop at one side of Petri dish with the Muller-Hinton agar (MHA) and incubated for 24 hours at 350 C in anaerobic conditions. It was supposed that bacteriocins would diffuse over the whole area of the agar media. On the next day S. pneumoniae, S. pyogenes, S. aureus, E. coli, P. aeruginosa and C. albicans clinical isolates were streaked at the clear side of MHA Petri dish. MHA Petri dish inoculated with SsK12 (one part) and with the respective clinical isolates (another part) streaked perpendicularly on the same day was used as the control. Results: There was no growth of S. pyogenes on the Petri dish with SsK12 daily culture; the growth of a few colonies of S. pneumonia was noted. The growth of S. aureus, E. coli, P. aeruginosa and C. albicans was noted along the inoculated streak. On the control Petri dish with simultaneous inoculating of the SsK12 strain and the test cultures, the growth of all the testes isolates was noted. Conclusions: (1) SsK12 possesses perfect antagonistic activity against S. pyogenes and good activity against S. pneumoniae. (2) There was no antagonistic activity of SsK12 against S. aureus, E. coli, P. aeruginosa and C. albicans. (3) SsK12 antagonistic properties make it possible to use this probiotic strain for prophylaxis of recurrent ENT infections.

Keywords: probiotics, SsK12, streptococcus salivarius K12, antagonistic activity

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Authors: Mrunalini Sonne

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Aquaculture uses a variety of broad spectrum antibiotics to manage and prevent a variety of diseases without understanding their mechanisms of action. This has led to water pollution in the modern world. The necessity for alternate control measures against bacterial illnesses in the aquaculture sector is highlighted by issues brought on by antibiotic-resistant bacteria and the dearth of effective control strategies. Bacteriophages (phages) have shown promise as therapeutic agents for the efficient management of bacterial infections in aquaculture. In the current study, a variety of investigations were conducted to determine if utilizing lytic phages to reduce Aeromonas spp. infection in fish aquaculture was appropriate. Motile Aeromonas septicaemia is a fish disease that has caused financial harm to the aquaculture sector. Currently, the production of aquaculture depends significantly on antibiotics, which adds to the worldwide problem of the rise of bacteria that are resistant to medicines and resistance genes. To decrease the usage of antibiotics in aquaculture systems, it is crucial to create efficient antibiotic substitutes. Bacteriophages are capable of acting as a natural antagonist, mostly because of their great specificity, capacity for self-replication, and ability to quickly eradicate dangerous bacteria. There is a need for research that goes beyond just isolating and characterising lytic bacteriophages to examine their morphology, stability, and efficacy in various environmental conditions. Bacteriophage (or phage) therapy is a promising technique to control dangerous microbes in farmed fish. More phage therapy research in aquaculture is required in order to effectively employ phage treatment to lessen infection in fish brought on by Aeromonas.

Keywords: aquaculture, bacteriophages, fish, freshwater

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Authors: Alazar Nebyou, Sujata Pandit

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Lactic acid bacteria (LAB) are essential ingredients in probiotic foods, intestinal microflora, and dairy products that are capable of coping up with harsh gastrointestinal tract conditions and are available in a variety of environments. The objective of this study is to evaluate the probiotic property of LAB isolated from bovine milk. Milk samples were collected from local dairy farms. Samples were obtained using sterile test tubes and transported to a laboratory in the icebox for further biochemical characterization. Preliminary physiological and biochemical identification of LAB isolates was conducted by growing on MRS agar after ten-fold serial dilution. Seven of the best isolates were selected for the evaluation of the probiotic property. The LAB isolates were checked for resistance to antibiotics and their antimicrobial activity by disc diffusion assay and agar well diffusion assay respectively. Bile salt hydrolase activity of isolates was studied by growing isolates in a BSH medium with bile salt. Cell surface property of isolates was assayed by studying their autoaggregation and coaggregation percentage with S. aerues. All isolates were found BSH positive. In addition, BCM2 and BGM1 were susceptible to all antibiotic disks except BBM1 which was resistant to all antibiotic disks. BCM1 and BGM1 had the highest autoaggregation and coaggregation potential respectively. Since all LAB isolates showed gastrointestinal tolerance and good cell surface property they could be considered as good potential probiotic candidates for treatment and probiotic starter culture preparation.

Keywords: probiotic, aggregation, lactic acid bacteria, antimicrobial activity

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Authors: Ngonidzashe Mangoma, Caroline Marigold Sebata

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The release of cyanide-rich effluents from gold mines, and other industries, into the environment, is a global concern considering the well-known metabolic effects of cyanide in all forms of life. Such effluents need to be treated to remove cyanide, among other pollutants, before their disposal. This study aimed at investigating the possible use of bacteria in the biological removal of cyanide from cyanide-rich effluents. Firstly, cyanide-degrading bacteria were isolated from gold mine effluents and characterised. The isolates were then tested for their ability to grow in the presence of cyanide and their tolerance to increasing levels of the compound. To evaluate each isolate’s cyanide-degrading activities, isolates were grown in the simulated and actual effluent, and a titrimetric method was used to quantify residual cyanide over a number of days. Cyanide degradation efficiency (DE) was then calculated for each isolate. Identification of positive isolates involved 16S rRNA gene amplification and sequence analysis through BLAST. Six cyanide-utilising bacterial strains were isolated. Two of the isolates were identified as Klebsiella spp. while the other two were shown to be different strains of Clostridium bifermentans. All isolates showed normal growth in the presence of cyanide, with growth being inhibited at 700 mg/L cyanide and beyond. Cyanide degradation efficiency for all isolates in the simulated effluent ranged from 79% to 97%. All isolates were able to remove cyanide from actual gold mine effluent with very high DE values (90 – 94%) being recorded. Isolates obtained in this study were able to efficiently remove cyanide from both simulated and actual effluent. This observation clearly demonstrates the feasibility of the biological removal of cyanide from cyanide-rich gold mine effluents and should, therefore, motivate research towards the possible large-scale application of this technology.

Keywords: cyanide effluent, bioremediation, Clostridium bifermentans, Klebsiella spp, environment

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