Search results for: fish spoilage bacteria

Authors: M. K. Sheikh, Y. M. Laghari., P. K. Lashari., N. T. Narejo

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The Indus River is one of the longest important rivers of the world in Asia that flows southward through Pakistan, merges into the Arabian Sea near the port city of Karachi in Sindh Province, and forms the Indus Delta. Fish are an important resource for humans worldwide, especially as food. In fish, healthy nutriments are present which are not found in any other meat source because it have a huge quantity of omega- 3 fatty acids, which are very essential for the human body. Ichthyologic surveys were conducted to explore the diversity of freshwater fishes, distribution, abundance and current status of the fishes at different spatial scale of the downstream, Indus River. Total eight stations were selected namely Railo Miyan (RM), Karokho (Kk), Khanpur (Kp), Mullakatiyar (Mk), Wasi Malook Shah (WMS), Branch Morie (BM), Sujawal (Sj) and Jangseer (JS). The study was carried in the period of January 2016 to December 2019 to identify River and biodiversity threats and to suggest recommendations for conservation. The data were analysed by different population diversity index. Altogether, 124 species were recorded belonging to 12 Orders and 43 Families from the downstream of Indus River. Among 124 species, 29% belong to high commercial value and 35% were trash fishes. 31% of fishes were identified as marine/estuarine origin (migratory) and 05% were exotic fish species. Perciformes is the most predominated order, contributing to 41% of families. Among 43 families, the family Cyprinidae was the richest family from all localities of downstream, represented by 24% of fish species demonstrating a significant dominance in the number of species. A significant difference was observed for species abundance in between all sites, the maximum abundance species were found at first location RM having 115 species and minimum observed at the last station JS 56 genera. In the recorded Ichthyofauna, seven groups were found according to the International Union for Conservation of Nature status (IUCN), where a high species ratio was collected, in Least Concern (LC) having 94 species, 11 were found as not evaluated (NE), whereas 8 was identified as near threatened (NT), 1 was recorded as critically endangered (CR), 11 were collected as data deficient (DD), and while 8 was observed as vulnerable (VU) and 3 endangered (EN) species. Different diversity index has been used extensively in environmental studies to estimate the species richness and abundance of ecosystems outputs of their wellness; a positive environment (biodiversity rich) with species at RM had an environmental wellness and biodiversity levels of 4.566% while a negative control environment (biodiversity poor) on last station JS had an environmental wellness and biodiversity levels of 3.931%. The status of fish biodiversity and river has been found under serious threat. Due to the lower diversity of fishes, it became not only venerable for fish but also risky for fishermen. Necessary steps are recommended to protect the biodiversity by conducting further conservative research in this area.

Keywords: ichthyofaunal biodiversity, threatened species, diversity index, Indus River downstream

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Authors: Dilek Zorlu Kaya, Sena Çenesiz, Utku Duran

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Infectious pancreatic necrosis virus is a disease of great concern in aquaculture, causing mortality of 80 - 90% of the stocks in salmonid production. We aimed to investigate the efficacy of quercetin on oxidant and antioxidant parameters of infectious pancreatic necrosis virus, which is important for fish farming and economy in vitro. Quercetin experimental model was used in the cell culture of Oncorhynchus mykiss infected with infectious pancreatic necrosis virus. Malondialdehyde, ceruloplasmin, total oxidant capacity, total antioxidant levels, and glutathione-peroxidase were measured in the samples. As a result of the study, it was observed that quercetin can minimize the damage caused by scavenging free radicals in cells infected with infectious pancreatic necrosis virus. Thus, we think that an important development can be achieved for fish farming and the economy.

Keywords: IPNV, oncorhynchus mykiss, TAS, TOS, quercetin

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Authors: S. F. Afzali, W. L. Wong

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The epizootic ulcerative syndrome (EUS) causes by the oomycete fungus, Aphanomyces invadans; known to be one of the infectious fish diseases for farmed and wild fishes in fresh and brackish-water from the Asia-pacific region, America and Africa. Although, EUS had been documented by the Office International des Epizooties (OIE) since 1995, hitherto, there is neither standard chemical agents that can be used for successful treatment of this destructive infection in the time of outbreak; nor available vaccine for prevention. Plant-based remedies in controlling fish diseases are gaining much attention recently as an alternative to chemical treatments, which possess negative effects to the environment and human. In present study, Sonneratia alba, a mangrove plant belongs to the Sonneratiaceae family, was screened in vitro and in vivo for its antifungal activity against A. invadans mycelium growth and its effects on fish innate immune system and disease resistant. The in vitro tests was performed using the disc diffusion methods with measurements of minimum inhibitory concentration (MIC) and inhibition zone. For in vivo study, the S. alba extract supplemented diets were administrated at 0.0, 1.0%, 3.0%, and 5.0% on healthy goldfish, Carassius auratus, which challenged with A. invadans zoospores (100 spores/ml). To compare the significant differences in the hematological and immunological parameters obtained from the experiments, the data were analysed using the SPSS. The methanol extract of S. alba effectively inhibited the mycelial growth of A. invadans at a minimum concentration of 1000 ppm for agar and filter paper diffusion experiments. In the agar diffusion test, 500 ppm of the extract inhibited the fungus mycelial growth up to 96 hours after exposure. The mycelial growth from the edge of the pre-inoculated A. invadans agar discs treated with S. alba extracts at concentrations of 100, 500 and 1000 ppm were 15, 8 and 0 mm respectively. The results of the filter paper disc test showed that the S. alba extract at its minimal inhibitory concentration (1000 ppm) has similar qualitative inhibitory effect as malachite green at 1 ppm and formalin at 250 ppm. According to the in vivo tests findings, in the infected fish fed with 3.0% and 5.0% supplementation diet, the numbers of white blood cell and myeloperoxidase activity significantly increased after the second week of treatment. Whilst the numbers of red blood cell significantly decreased in the infected fish fed with 0.0 and 1.0% supplementation diet. After the third week of feeding, significant increases in the total protein, albumin level, lysozyme activity were recorded in the infected fish fed with 3.0% and 5.0% supplementation diet. Also, the enriched diets increased the survival rate as compared to the untreated group that suffered from 90% mortality. The present study indicated that S. alba extract may inhibit the mycelial growth of A. invadans effectively, suggesting an alternative to other chemotherapeutic agents, which brought much environmental and health concerns to the public, for EUS treatment.

Keywords: fungal pathogen, goldfish, organic extract, treatment

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Authors: Aliasghar Golmohammadian, Sona Rostampour Yasouri

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One of the most important intestinal pathogenic strains is E. coli O₁₅₇:H₇. This pathogenic bacterium is transmitted to humans through water and food. E. coli O₁₅₇:H₇ is the main cause of Hemorrhagic colitis (HC), Hemolytic Uremic Syndrome (HUS), Thrombotic Thrombocytopenic Purpura (TTP) and in some cases death. Since E. coli O₁₅₇:H₇ can be transmitted through the consumption of different foods, including vegetables, agricultural products, and fresh dairy products, this study aims to identify and isolate E. coli O₁₅₇:H₇ from wastewater by PCR technique. One hundred twenty samples of water and wastewater were collected by Falcom Sterile from Shahrood and Neka cities. The samples were checked for colony formation after appropriate centrifugation and cultivation in the specific medium of Sorbitol MacConkey Agar (SMAC) and other diagnostic media of E. coli O₁₅₇:H₇. Also, the plates were observed macroscopically and microscopically. Then, the necessary phenotypic tests were performed on the colonies, and finally, after DNA extraction, the PCR technique was performed with specific primers related to rfbE and stx2 genes. The number of 5 samples (6%) out of all the samples examined were determined positive by PCR technique with observing the bands related to the mentioned genes on the agarose gel electrophoresis. PCR is a fast and accurate method to identify the bacteria E. coli O₁₅₇:H₇. Considering that E. coli bacteria is a resistant bacteria and survives in water and food for weeks and months, the PCR technique can provide the possibility of quick detection of contaminated water. Moreover, it helps people in the community control and prevent the transfer of bacteria to healthy and underground water and agricultural and even dairy products.

Keywords: E. coli O₁₅₇:H₇, PCR, water, wastewater

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Authors: Salina Saddick, Mohamed Afifi, Osama Abuznadah

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This study was carried out to determine the median lethal concentrations (LC50) of Zinc nanoparticles (ZnNPs) on Oreochromis niloticus and Tilapia zillii. The biochemical and molecular potential effects of ZnNPs (500 and 2000 μg L−1) on the antioxidant system in the brain tissue of O. niloticus and T. zillii were investigated. Four hundred fish were used for acute and sub-acute studies. ZnNP LC50 concentrations were investigated in O. niloticus and T. zillii. The effect of 500 and 2000 μg L−1 ZnNPs on brain antioxidants of O. niloticus and T. zillii was investigated. The result indicated that 69 h LC50 was 5.5 ± 0.6 and 5.6 ± 0.4 for O. nilotica and T. zillii, respectively. Fish exposed to 500 μg L−1 ZnNPs showed a significant increase in reduced glutathione (GSH), total glutathione (tGSH) levels, superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) activity and gene expression. On the contrary, malondialdehyde (MDA) levels significantly decreased. Meanwhile, fish exposed to 2000 μg L−1 ZnNPs showed a significant decrease of GSH, tGSH levels, SOD, CAT, GR, GPx and GST activity and gene expression. On the contrary, MDA levels significantly increased. It was concluded that, the 96 h LC50 of ZnNPs was 5.5 ± 0.6 and 5.6 ± 0.4 for O. nilotica and T. zillii, respectively. ZnNPs in exposure concentrations of 2000 μg/L induced a deleterious effect on the brain antioxidant system of O. nilotica and T. zillii. In contrast, ZnNPs in exposure concentrations of 500 μg L−1 produced an inductive effect on the brain antioxidant system of O. nilotica and T. zillii.

Keywords: ZnNPs, LC50, antioxidants, O. nilotica

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Authors: H. Nabahat, E. Amiri, F. AzaditalabDavoudabadi, N. Zaeri

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Tooth decay is one of the most common forms of oral and dental illness in the world, which causes huge costs of treatment, especially in high-risk groups such as people with oral dry mouth, prevention and control of it are very important. The use of traditional treatments such as extraction of drugs from medicinal plants is of paramount importance to Iran and the international community as well. The present study was conducted with the aim of investigating the antibacterial effect of the extract of Salvia officinalis and Mentha pulegium, which are the most commonly used drugs in the treatment of oral and teeth bacterial (Streptococcus mutant, Lactobacillus rhamnosis, and Actinomyces viscosis) in vitro method. In this experimental study, two herbs of Salvia and Mentha were prepared by maceration of hydroalcoholic extract, and the antibacterial effect was evaluated by broth macro dilution on streptococcal mutagen bacteria, lactobacillus rhamnosis, and viscose actinomycosis. The results were analyzed by the Whitney Mann test (P > 0.05). The results showed that the minimum inhibitory concentration (MIC) of the salmonella extract for Streptococcus mutan were 6.25 and 12.5 μg/ml, respectively, for lactobacillus of 1.56 and 3.12 μg/ml, respectively, and for actinomycosis viscose, The order of 12.5 and 100 μg/ml was obtained. As a result, broth macro dilution showed that both extracts of Salvia and Mentha had an inhibitory effect on all three species of bacteria. This effect for Salvia was significantly (P < 0.05) more than Mentha and was within the concentration range of both the extracts and had a bactericidal effect on all three bacteria.

Keywords: antibacterial effect, dental bacteria, herbal extracts , salvia officinalis, mentha pulegium

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Authors: B. Tayebani, N. Hosseinibalam, D. Mostofinejad

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Interest in using bacteria in cement materials due to its positive influences has been increased. Cement materials such as mortar and concrete basically suffer from higher porosity and water absorption compared to other building materials such as steel materials. Because of the negative side-effects of certain chemical techniques, biological methods have been proposed as a desired and environmentally friendly strategy for reducing concrete porosity and diminishing water absorption. This paper presents the results of an experimental investigation carried out to evaluate the influence of Sporosarcina pasteurii bacteria on the behaviour of two types of concretes (light weight aggregate concrete and normal weight concrete). The resistance of specimens to water penetration by testing water absorption and evaluating the electrical resistance of those concretes was examined and compared. As a conclusion, 20% increase in electrical resistance and 10% reduction in water absorption of lightweight aggregate concrete (LWAC) and for normal concrete the results show 7% decrease in water absorption and almost 10% increase in electrical resistance.

Keywords: bacteria, biological method, normal weight concrete, lightweight aggregate concrete, water absorption, electrical resistance

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Authors: Hacer Yeldan, Erhan Akamca, Sedat Gündogdu

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Length-Weight and Length-length relationship were estimated of 14 species Sparidae (Boops boops, Diplodus annularis, Diplodus cervinus, Dipladus puntazzo, Diplodus sargus, Diplodus vulgaris, Lithognathus mormyrus, Oblada melanura, Pagellus acarne, Pagellus erythrinus, Pagrus auriga, Pagrus caeruleostictus, Sarpa salpa, Sparus aurata) sampled from in the Northeastern Mediterranean Sea coast of Turkey, Iskenderun Bay. Samples were collected from July 2014 to June 2015, using bottom trawl and trammel net into three different depth; 0-10 m, 10-20 m, 20-50m. Length-length relationships were determined size measurements: standard length (SL) and fork length (FL) to total length (TL) for fish species. The relationships between TL, FL and TL, SL were all linear. The values of the exponent b of the length-weight relationships ranged between 2.685 and 3.473. The type of growth for fish species was algometric growth.

Keywords: Sparidae, Iskenderun bay, length-length, length-weight relationships

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Authors: Negar Zaheddoust, Negin Zaheddoust, Abbas Asoudeh-Fard

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Probiotic bacteria have been employed as a novel and less side-effect strategy for anticancer therapy. Since the oral cavity is a host for probiotic and pathogen bacteria to colonize, more investigation is needed to evaluate the effectiveness of this novel adjunctive treatment for oral cancer. We considered Lactobacillus plantarum as a probiotic and Streptococcus mutans as a pathogen bacterium in our study. The aim of this study is to examine the effect of Lactobacillus plantarum and Streptococcus mutans on Casp3, AKT / PTEN, and MAPK signaling pathway, which is involved in apoptosis or survival of oral cancer KB cells. On the other hand, to study the effects of these bacteria on normal cells, we used HUVEC cells. The KB and HUVEC cell lines were co-cultured with Lactobacillus plantarum and Streptococcus mutans isolated from traditional Iranian dairy and dental plaque, respectively. The growth-inhibitory effects of these two bacteria on KB and HUVEC cells were determined by (3-(4, 5-dimethylthiazolyl-2)-2,5diphenyltetrazolium bromide) MTT assay. MTT results demonstrated that the proliferation of KB cells was affected in a time, dose, and strain-dependent manner. In the following, the examination of induced apoptosis or necrosis in co-cultured KB cells with the best IC50 concentration of the Lactobacillus plantarum and Streptococcus mutans will be analyzed by FACS flow cytometry, and the changes in gene expression of Casp3, AKT / PTEN, MAPK genes will be evaluated using real-time polymerase chain reaction.

Keywords: cancer therapy, induced apoptosis, oral cancer, probiotics

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Authors: Rashad R. Al-Hindi

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The aim of this study was to isolate and identify lactic acid bacteria and evaluate their therapeutic and food preservation importance. Ninety-three suspected lactic acid bacteria (LAB) were isolated from thirteen different raw and fermented milk of indigenous sources in the Kingdom of Saudi Arabia. The identification of forty-six selected LAB strains and genetic relatedness were performed based on 16S rDNA gene sequence comparison. The LAB counts in certain samples were higher under microaerobic than anaerobic conditions. The identified LAB belonged to genera Enterococcus (16 strains), Lactobacillus (9 strains), Weissella (10 strains), Streptococcus (8 strains) and Lactococcus (3 strains). Phylogenetic tree generated from the full-length (~1.6 kb) sequences confirmed previous findings. Utilization of shorter 16S rDNA sequences (~1.0 kb) also discriminated among strains of which V2 region was the most effective. None of the strains exhibited resistance to clinically relevant antibiotics or undesirable hemolytic activity, while they differed in other probiotic characteristics, e.g., tolerance to acidic pH, resistance to bile salt, and antibacterial activity. In conclusion, the isolates Lactobacillus casei MSJ1, Lactobacillus casei Dwan5, Lactobacillus plantarum EyLan2 and Enterococcus faecium Gail-BawZir8 are likely the best probiotic LAB and we speculate that studying the synergistic effects of bacterial combinations might result in the occurrence of more effective probiotic potential. We argue that the raw and fermented milk of animals hosted in Saudi Arabia, especially stirred yogurt (Laban) made from camel milk, are rich in LAB with promising probiotics potential.

Keywords: fermented foods, lactic acid bacteria, probiotics, Saudi Arabia

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Authors: Pankaj Dinesh Baviskar

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This study examines the impact of curcumin-loaded nano-spheres in the form of emulsions on fish eye cataracts. Curcumin nanoemulsions were prepared by using phenethyl isothiocyanate. Nanoemulsions were synthesized by ultrasound-assisted method at 150 Watt. A zeta potential measurement for curcumin-loaded nanoemulsions was found to be -30.7eV, -13.4eV, and -9.55eV, and particle size was found to be 149.3 nm, 245.3 and nm 403.5 nm using particle size analyzer respectively for different conditions. The surface morphology of nano-spheres was examined by FE-SEM analysis. The zeta potential measured indicates its stability for corresponding nano-spheres. The anti-cataract application was studied by using isolated fish eye lenses. The cataract was induced using high glucose concentrated solution. The biochemical parameters in the form of reduced glutathione were measured to interpret the anti-cataract ability of curcumin-loaded nanoemulsions.

Keywords: curcumin, nano, cataract, nanoemulsion

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Authors: Tsige Reda

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Essential oil of Thymus vulgaris was extracted by means of hydro-distillation. This study was done to investigate the chemical composition, antibacterial and antioxidant activities. The chemical composition of the essential oils was determined using gas chromatography coupled to mass spectroscopy (GC-MS). Using disc diffusion assay the antibacterial activity was assessed on one Gram-positive bacteria and one Gram-negative bacteria. The percentage oil yield of the essential oil was found to be 0.97 ± 0.08% (w/w) with yellow color. The physicochemical constants of the oil were also noted. The phytochemical screening of the plant extract revealed the presence of tannins, saponins, phenol, flavonoids, terpenoids, steroids and alkaloids. A total of 18 chemical constituents were identified by Gas Chromatography-Mass Spectroscopy analysis representing 100% of the total essential oil of Thymus vulgaris, with thymol (31.977%), o-cymene (29.992%), and carvacrol (14.541%). Previous studies have revealed that the thymol, o-cymen and carvacrol components of Thymus vulgaris are responsible for their biological activities. Thymus vulgaris have been used traditionally to treat a wide variety of infections. Based on the extensive use and lack of scientific evidence, a study was embarked upon to determine its bioactivity. The essential oil of Thymus vulgaris leaves exhibited higher activity towards the Gram-positive bacteria (Staphylococcus aurous) than the Gram-negative bacteria (Escherichia coli) and also has good antioxidant activity, and can be used medicinal and therapeutic applications. This activity may be due to the high amount of thymol, o-cymen and carvacrol.

Keywords: hydro-distillation, Thymus vulgaris, essential oil composition, phytochemical screening, physicochemical constants, antioxidant activity, antibacterial activity

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Authors: Thangavelu Muthukumar, Thotapalli Parvathaleswara Sastry

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Collagen is the most abundantly available connective tissue protein, which is being used as a biomaterial for various biomedical applications. Presently, fish wastes are disposed improperly which is causing serious environmental pollution resulting in offensive odour. Fish scales are promising source of Type I collagen. Medicinal plants have been used since time immemorial for treatment of various ailments of skin and dermatological disorders especially cuts, wounds, and burns. Developing biomaterials from the natural sources which are having wound healing properties within the search of a common man is the need of hour, particularly in developing and third world countries. With these objectives in view we have developed a wound dressing material containing fish scale collagen (FSC) incorporated with Macrotyloma uniflorum plant extract (PE). The wound dressing composite was characterized for its physiochemical properties using conventional methods. SEM image revealed that the composite has fibrous and porous surface which helps in transportation of oxygen as well as absorbing wound fluids. The biomaterial has shown 95% biocompatibility with required mechanical strength and has exhibited antimicrobial properties. This biomaterial has been used as a wound dressing material in experimental wounds of rats. The healing pattern was evaluated by macroscopic observations, panimetric studies, biochemical, histopathological observations. The results showed faster healing pattern in the wounds treated with CSPE compared to the other composites used in this study and untreated control. These experiments clearly suggest that CSPE can be used as wound/burn dressing materials.

Keywords: collagen, wound dressing, Macrotyloma uniflorum, burn dressing

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Authors: M. E. Abalaka, S. Y. Daniyan, S. O. Adeyemo, D. Damisa

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Gold nanoparticles (AuNPs) have gained increasing interest in recent times. This is greatly due to their special features, which include unusual optical and electronic properties, high stability and biological compatibility, controllable morphology and size dispersion, and easy surface functionalization. In typical synthesis, AuNPs were produced by reduction of gold salt AuCl4 in an appropriate solvent. A stabilizing agent was added to prevent the particles from aggregating. The antibacterial activity of different sizes of gold nanoparticles was investigated against Staphylococcus aureus, Salmonella typhi and Pseudomonas pneumonia using the disk diffusion method in a Müeller–Hinton Agar. The Au-NPs were effective against all bacteria tested. That the Au-NPs were successfully synthesized in suspension and were used to study the antibacterial activity of the two medicinal plants against some bacterial pathogens suggests that Au-NPs can be employed as an effective bacteria inhibitor and may be an effective tool in medical field. The study clearly showed that the Au-NPs exhibiting inhibition towards the tested pathogenic bacteria in vitro could have the same effects in vivo and thus may be useful in the medical field if well researched into.

Keywords: gold nanoparticles, Gomphrena celesioides, Prunus amygdalus, pathogens

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Authors: Jeet Chakraborty, Sanjay Dutta

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Antibiotic resistance by bacteria has proved to be a severe threat to mankind in recent times, and this fortifies an urgency to design and develop potent antibacterial small molecules/compounds with nonconventional mechanisms than the conventional ones. DNA carries the genetic signature of any organism, and bacteria maintain their genomic DNA inside the cell in a well-regulated compact form with the help of various nucleoid associated proteins like HU, HNS, etc. These proteins control various fundamental processes like gene expression, replication, etc., inside the cell. Alteration of the native DNA structure of bacteria can lead to severe consequences in cellular processes inside the bacterial cell that ultimately result in the death of the organism. The change in the global DNA structure by small molecules initiates a plethora of cellular responses that have not been very well investigated. Echinomycin and Triostin-A are biologically active Quinoxaline small molecules that typically consist of a quinoxaline chromophore attached with an octadepsipeptide ring. They bind to double-stranded DNA in a sequence-specific way and have high activity against a wide variety of bacteria, mainly against Gram-positive ones. To date, few synthetic quinoxaline scaffolds were synthesized, displaying antibacterial potential against a broad scale of pathogenic bacteria. QNOs (Quinoxaline N-oxides) are known to target DNA and instigate reactive oxygen species (ROS) production in bacteria, thereby exhibiting antibacterial properties. The divergent role of Quinoxaline small molecules in medicinal research qualifies them for the evaluation of their antimicrobial properties as a potential candidate. The previous study from our lab has given new insights on a 6-nitroquinoxaline derivative 1d as an intercalator of DNA, which induces conformational changes in DNA upon binding.7 The binding event observed was dependent on the presence of a crucial benzyl substituent on the quinoxaline moiety. This was associated with a large induced CD (ICD) appearing in a sigmoidal pattern upon the interaction of 1d with dsDNA. The induction of DNA superstructures by 1d at high Drug:DNA ratios was observed that ultimately led to DNA condensation. Eviction of invitro-assembled nucleosome upon treatment with a high dose of 1d was also observed. In this work, monoquinoxaline derivatives of 1d were synthesized by various modifications of the 1d scaffold. The set of synthesized 6-nitroquinoxaline derivatives along with 1d were all subjected to antibacterial evaluation across five different bacteria species. Among the compound set, 3a displayed potent antibacterial activity against Staphylococcus aureus bacteria. 3a was further subjected to various biophysical studies to check whether the DNA structural alteration potential was still intact. The biological response of S. aureus cells upon treatment with 3a was studied using various cell biology processes, which led to the conclusion that 3d can initiate DNA damage in the S. aureus cells. Finally, the potential of 3a in disrupting preformed S.aureus and S.epidermidis biofilms was also studied.

Keywords: DNA structural change, antibacterial, intercalator, DNA superstructures, biofilms

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Authors: George Abu-Aqil, Leah Tsror, Elad Shufan, Shaul Mordechai, Mahmoud Huleihel, Ahmad Salman

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Pectobacterium and Dickeya spp which negatively affect a wide range of crops are the main causes of the aggressive diseases of agricultural crops. These aggressive diseases are responsible for a huge economic loss in agriculture including a severe decrease in the quality of the stored vegetables and fruits. Therefore, it is important to detect these pathogenic bacteria at their early stages of infection to control their spread and consequently reduce the economic losses. In addition, early detection is vital for producing non-infected propagative material for future generations. The currently used molecular techniques for the identification of these bacteria at the strain level are expensive and laborious. Other techniques require a long time of ~48 h for detection. Thus, there is a clear need for rapid, non-expensive, accurate and reliable techniques for early detection of these bacteria. In this study, infrared spectroscopy, which is a well-known technique with all its features, was used for rapid detection of Pectobacterium and Dickeya spp. at the strain level. The bacteria were isolated from potato plants and tubers with soft rot symptoms and measured by infrared spectroscopy. The obtained spectra were analyzed using different machine learning algorithms. The performances of our approach for taxonomic classification among the bacterial samples were evaluated in terms of success rates. The success rates for the correct classification of the genus, species and strain levels were ~100%, 95.2% and 92.6% respectively.

Keywords: soft rot enterobacteriaceae (SRE), pectobacterium, dickeya, plant infections, potato, solanum tuberosum, infrared spectroscopy, machine learning

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Authors: M. Thuanthong, W. Youravong, N. Sirinupong

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Nile tilapia (Oreochromis niloticus) is one of fish species cultured in Thailand with a high production volume. A lot of skin is generated during fish processing. In addition, there are many research reported that fish skin contains abundant of collagen. Thus, the use of Nile tilapia skin as collagen source can increase the benefit of industrial waste. In this study, Acid soluble collagen (ASC) was extracted at 5, 15 or 25 ˚C with 0.5 M acetic acid then the acid was removed out and collagen was concentrated by ultrafiltration-diafiltration (UFDF). The triple helix collagen from UFDF process was used as substrate to produce collagen peptides by alcalase hydrolysis in an enzymatic membrane reactor (EMR) coupling with 1 kDa molecular weight cut off (MWCO) polysulfone hollow fiber membrane. The results showed that ASC extracted at high temperature (25 ˚C) with 0.5 M acetic acid for 5 h still preserved triple helix structure. In the UFDF process, the acid removal was higher than 90 % without any effect on ASC properties, particularly triple helix structure as indicated by circular dichroism spectrum. Moreover, Collagen from UFDF was used to produce collagen peptides by EMR. In EMR, collagen was pre-hydrolyzed by alcalase for 60 min before introduced to membrane separation. The EMR operation was operated for 10 h and provided a good of protein conversion stability. The results suggested that there is a successfulness of UF in application for acid removal to produce ASC with desirable preservation of its quality. In addition, the EMR was proven to be an effective process to produce low molecular weight peptides with ACE-inhibitory activity properties.

Keywords: acid soluble collagen, ultrafiltration-diafiltration, enzymatic membrane reactor, ace-inhibitory activity

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Authors: Etleva Hamzaraj, Eva Kica, Anila Paparisto, Pranvera Lazo

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Worldwide demographic developments of recent decades have been associated with negative environmental consequences. For this reason, there is a growing interest in assessing the state of natural ecosystems or assessing human impact on them. In this respect, this study aims to evaluate the change in water quality of the Mat River for a period of about ten years to highlight human impact. In one year, period of study, several biological and environmental parameters are determined to evaluate river water quality, and the data collected are compared with those of a similar study in 2007. Samples are collected every month in five stations evenly distributed along the river. Total coliform bacteria, the number of heterotrophic bacteria in water, and benthic macroinvertebrates are used as biological parameters of water quality. The most probable number index is used for evaluation of total coliform bacteria in water, while the number of heterotrophic bacteria is determined by counting colonies on plates with Plate Count Agar, cultivated with 0.1 ml sample after series dilutions. Benthic macroinvertebrates are analyzed by the number of individuals per taxa, the value of biotic index, EPT Richness Index value and tolerance value. Environmental parameters like pH, temperature, and electrical conductivity are measured onsite. As expected, the bacterial load was higher near urban areas, and the pollution increased with the course of the river. The maximum concentration of fecal coliforms was 1100 MPN/100 ml in summer and near the most urbanized area of the river. The data collected during this study show that after about ten years, there is a change in water quality of Mat River. According to a similar study carried out in 2007, the water of Mat River was of ‘excellent’ quality. But, according to this study, the water was classified as of ‘excellent’ quality only in one sampling site, near river source, while in all other stations was of ‘good’ quality. This result is based on biological and environmental parameters measured. The human impact on the quality of water of Mat River is more than evident.

Keywords: water quality, coliform bacteria, MPN index, benthic macroinvertebrates, biotic index

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Authors: Elena Carcano, James Ball, Betty Tiko

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Carrying capacity refers to the maximum population that a given level of resources can sustain over a specific period. In undisturbed environments, the maximum population is determined by the availability and distribution of resources, as well as the competition for their utilization. This information is typically obtained through long-term data collection. In regulated environments, where resources are artificially modified, populations must adapt to changing conditions, which can lead to additional challenges due to fluctuations in resource availability over time and throughout development. An example of this is observed in hydropower plants, which alter water flow and impact fish migration patterns and behaviors. To assess how fish species can adapt to these changes, specialized surveys are conducted, which provide valuable information on fish populations, sample sizes, and density before and after flow modifications. In such situations, it is highly recommended to conduct hydrological and biological monitoring to gain insight into how flow reductions affect species adaptability and to prevent unfavorable exploitation conditions. This analysis involves several planned steps that help design appropriate hydropower production while simultaneously addressing environmental needs. Consequently, the study aims to strike a balance between technical assessment, biological requirements, and societal expectations. Beginning with a small hydro project that requires restoration, this analysis focuses on the lower tail of the Flow Duration Curve (FDC), where both hydrological and environmental goals can be met. The proposed approach involves determining the threshold condition that is tolerable for the most vulnerable species sampled (Telestes Muticellus) by identifying a low flow value from the long-term FDC. The results establish a practical connection between hydrological and environmental information and simplify the process by establishing a single reference flow value that represents the minimum environmental flow that should be maintained.

Keywords: carrying capacity, fish bypass ladder, long-term streamflow duration curve, eta-beta method, environmental flow

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Authors: Ali Seirafi

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The purpose of this study was to investigate the effect and application of starch nanocrystals on physicochemical and microbial properties in the industrial production of probiotic yogurt. In this study, probiotic yoghurt was manufactured by industrial method with the optimization and control of the technological factors affecting the probabilistic biomass, using probiotic bacteria Lactobacillus acidophilus and Bifidobacterium bifidum with commonly used yogurt primers. Afterwards, the effects of different levels of fat (1.3%, 2.5 and 4%), as well as the effects of various perbiotic compounds include starch nanocrystals (0.5%, 1 and 1.5%), galactolegalosaccharide (0.5% 1 and 1.5%) and fructooligosaccharide (0.5%, 1 and 1.5%) were evaluated. In addition, the effect of packaging (polyethylene and glass) was studied, while the effect of pH changes and final acidity were studied at each stage. In this research, all experiments were performed in 3 replications and the results were analyzed in a completely randomized design with SAS version 9.1 software. The results of this study showed that the addition of starch nanocrystal compounds as well as the use of glass packaging had the most positive effects on the survival of Lactobacillus acidophilus bacteria and the addition of nano-crystals and the increase in the cooling rate of the product, had the most positive effects on the survival of bacteria Bifidobacterium bifidum.

Keywords: Bifidobacterium bifidum, Lactobacillus acidophilus, prebiotics, probiotic yogurt

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Authors: Preeyarach Whisetkhan, Malai Taweechotipatr, Ulisa Pachekrepapol

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Elevated level of blood cholesterol or hypercholesterolemia may lead to atherosclerosis and poses a major risk for cardiovascular diseases. Probiotics play a crucial role in human health, and probiotic bacteria that possesses bile salt hydrolase (BSH) activity can be used to lower cholesterol level of the host. The aim of this study was to investigate whether lactic acid bacteria (LAB) isolated from traditional Thai fermented foods were able to exhibit bile salt hydrolase activity and their use in fermented milk. A total of 28 isolates were tested for BSH activity by plate method on MRS agar supplemented with 0.5% sodium salt of taurodeoxycholic acid and incubated at 37°C for 48 h under anaerobic condition. The results showed that FN1-1 and FN23-3 isolates possessed strong BSH activity. FN1-1 and FN23-3 isolates were then identified for phenotype, biochemical characteristics, and genotype (16S rRNA sequencing). FN1-1 isolate showed 99.92% similarity to Lactobacillus pentosus DSM 20314(T), while FN23-3 isolate showed 99.94% similarity to Enterococcus faecium CGMCC1.2136 (T). Lactobacillus pentosus FN1-1 and Enterococcus faecium FN23-3 were tolerant of pH 3-4 and 0.3 and 0.8% bile. Bacterial count and pH of milk fermented with Lactobacillus pentosus FN1-1 at 37°C and 43°C were investigated. The results revealed that Lactobacillus pentosus FN1-1 was able to grow in milk, which led to decrease in pH level. Fermentation at 37°C resulted in faster growth rate than at 43 °C. Lactobacillus pentosus FN1-1 was a candidate probiotic to be used in fermented milk products to reduce the risk of high-cholesterol diseases.

Keywords: probiotics, lactic acid bacteria, bile salt hydrolase, cholesterol

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Authors: Oluyemi O. Awolusi, Abimbola M. Enitan, Sheena Kumari, Faizal Bux

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Nitrification is essential to biological processes designed to remove ammonia and/or total nitrogen. It removes the excess nitrogenous compound in wastewater which could be very toxic to the aquatic fauna or cause a serious imbalance of such aquatic ecosystem. Efficient nitrification is linked to an in-depth knowledge of the structure and dynamics of the nitrifying community structure within the wastewater treatment systems. In this study, molecular technique was employed for characterizing the microbial structure of activated sludge [ammonia oxidizing bacteria (AOB) and nitrite oxidizing bacteria (NOB)] in a municipal wastewater treatment with intention of linking it to the plant efficiency. PCR-based phylogenetic analysis was also carried out for. The average operating and environmental parameters, as well as specific nitrification rate of a plant, was investigated during the study. During the investigation, the average temperature was 23±1.5oC. Other operational parameters such as mixed liquor suspended solids and chemical oxygen demand inversely correlated with ammonia removal. The dissolved oxygen level in the plant was constantly lower than the optimum (between 0.24 and 1.267 mg/l) during this study. The plant was treating wastewater with the influent ammonia concentration of 31.69 and 24.47 mg/l. The influent flow rates (ML/day) was 96.81 during the period. The dominant nitrifiers include: Nitrosomonas spp. Nitrobacter spp. and Nitrospira spp. The AOB had a correlation with nitrification efficiency and temperature. This study shows that the specific ammonia oxidizing rate and the specific nitrate formation rates can serve as a good indicator of the plant overall nitrification performance.

Keywords: Ammonia monooxygenase α-subunit gene, amoA, ammonia-oxidizing bacteria, AOB, nitrite-oxidizing bacteria, NOB, specific nitrification rate

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Authors: Abdurrahim A. Elouzi, Abdurrauf M. Gusbi, Ali M. Elgerbi

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Antibiotic resistance has become a global public-health problem, thus it is imperative that new antibiotics continue to be developed. Major problems are being experienced in human medicine from antibiotic resistant bacteria. Moreover, no new chemical class of antibiotics has been introduced into medicine in the past two decades. The aim of the current study presents experimental results that evaluate the capability of bio surfactant rhamnolipid on enhancing the efficacy of hydrophobic antibiotics. Serial dilutions of azithromycin and clarithromycin were prepared. A bacterial suspension (approximately 5 X 105 CFU) from an overnight culture in MSM was inoculated into 20 ml sterile test tube each containing a serial 10-fold dilution of the test antibiotic(s) in broth with or without 200 mgL-1 rhamnolipid. The tubes were incubated for 24 h with vigorous shaking at 37°C. Antimicrobial activity in multiple antibiotic-resistant gram-negative bacteria pathogens and gram-positive bacteria were assessed using optical density technique. The results clearly demonstrated that the presence of rhamnolipid significantly improved the efficiency of both antibiotics. We hypothesized that the addition of rhamnolipid at low concentration, causes release of LPS which results in an increase in cell surface hydrophobicity. This allows increased association of cells with hydrophobic antibiotics resulting in increased cytotoxicity rates.

Keywords: hydrophobic antibiotics, biosurfactant, rhamnolipid, azithromycin, clarithromycin

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Authors: B. Chandar, M. K. Ramasamy, P. Madasamy

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The discovery and development of newer antibiotics are limited with the increase in resistance of such multi-drug resistant bacteria creating the need for alternative new therapeutic agents. The recently discovered New Delhi Metallo-betalactamase-1 (NDM-1), which confers antibiotic resistance to most of the currently available β-lactams, except colistin and tigecycline, is a global concern. Several antibacterial drugs approved are natural products or their semisynthetic derivatives, but plant extracts remain to be explored to find molecules that are effective against NDM-1 bacteria. Therefore, it is necessary to explore the possibility of finding new and effective antibacterial compounds against NDM-1 bacteria. In the present study, we have screened a diverse set South Indian plant species, and report most plant species as a potential source for antimicrobial compounds against NDM-1 bacteria. Ethanol extracts from the leaves of taxonomically diverse South Indian medicinal plants were screened for antibacterial activity against NDM-1 E. coli using streak plate method. Among the plant screened against NDM-1 E. coli, the ethanol extracts from many plant extracts showed minimum bactericidal concentration between 5 and 15 mg /ml and MIC between 2.54 and 5.12 mg/ml. These extracts also showed a potent synergistic effect when combined with antibiotics colistin and tetracycline. Combretum albidum that was effective was taken for further analysis. At 5mg/L concentration, these extracts inhibited the NDM-1 enzyme in vitro, and residual activity for Combretum albidum was 33.09%. Treatment of NDM-1 E. coli with the extracts disrupted the cell wall integrity and caused 89.7% cell death. The plant extract of Combretum albidum that was effective was subjected to fractionation and the fraction was further subjected to HPLC, LC-MS for identification of antibacterial compound.

Keywords: antibacterial activity, combretum albidum, Escherichia coli, NDM-1

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Authors: Anna D. Kotrova, Alexandr N. Shishkin, Elena I. Ermolenko

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The aim. To study the quantitative and qualitative colon bacteria ratio from patients with metabolic syndrome. Materials and methods. Fecal samples from patients of 2 groups were identified and analyzed: the first group was formed by patients with metabolic syndrome, the second one - by healthy individuals. The metagenomics method was used with the analysis of 16S rRNA gene sequences. The libraries of the variable sites (V3 and V4) gene 16S RNA were analyzed using the MiSeq device (Illumina). To prepare the libraries was used the standard recommended by Illumina, a method based on two rounds of PCR. Results. At the phylum level in the microbiota of patients with metabolic syndrome compared to healthy individuals, the proportion of Tenericutes was reduced, the proportion of Actinobacteria was increased. At the genus level, in the group with metabolic syndrome, relative to the second group was increased the proportion of Lachnospira. Conclusion. Changes in the colon bacteria ratio in the gut microbiota of patients with metabolic syndrome were found both at the type and the genus level. In the metabolic syndrome group, there is a decrease in the proportion of bacteria that do not have a cell wall. To confirm the revealed microbiota features in patients with metabolic syndrome, further study with a larger number of samples is required.

Keywords: gut microbiota, metabolic syndrome, metagenomics, tenericutes

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Authors: I. Kamika, S. Azizi, M. Tekere

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Nanotechnology promises significant improvements of advanced materials and manufacturing techniques with a vast range of applications, which are critical for the future competitiveness of national industries. The manipulations and productions of materials, whilst, controlling the optical properties and surface area to a nanosize scale enabled a birth of a new field known as nanotechnology. However, their rapidly developing industry raises concerns about the environmental impacts of nanoparticles, as their effects on functional bacterial community in wastewater treatment remain unclear. The present research assessed the impact of cerium Oxide nanoparticles (nCeO) on the bacterial microbiome of an activated sludge system, which influenced its performance of this system on nutrient removal. Out of 15875 reads sequenced, a total of 13133 reads were non-chimeric. The wastewater samples were more dominant to the unclassified bacteria (51.07% of bacteria community) followed with the classified bacteria (48.93). Proteobacteria was the most dominant phylum in both classified and unclassified bacteria, whereas 18% of bacteria could even not be assigned a phylum and remained unclassified suggesting hitherto vast untapped microbial diversity. The bacterial operational taxonomic units (OTUs) ranged from 1014 to 2629 over the experimental period. The denitrification related species including Diaphorobacter species, Thauera species and those in the Sphaerotilus and Leptothrix group were found to be inhibited in a high concentration of CeO-NP. The diversity indices suggested that the bacterial community inhabiting the wastewater samples were less diverse as the concentration of CeO increases. The canonical correspondence analysis (CCA) results highlighted that the bacterial community variance had the strongest relationship with water temperature, conductivity, pH, and dissolved oxygen (DO) content as well as nCeO. The results provided the relationships between the microbial community and environmental variables in the wastewater samples.

Keywords: bacterial community, next generation, cerium oxide, wastewater, activated sludge, nanoparticles, nanotechnology

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Authors: Yael Shai, Dror L. Angel, Dror Zurel, Peleg Astrahan, Maxim Rubin-Blum, Eyal Rahav

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The high demand for oil and its by-products is symptomatic of the 21st century and occasionally leads to oil spills and pollution of coastal waters. Marine oil pollution may originate from a variety of sources -urban runoff, tanker cleaning, drilling activities, and oil spills. These events may release large amounts of highly toxic polycyclic aromatic hydrocarbons (PAHs) and other pollutants to coastal water, thereby threatening local marine life. Here, we investigated the effects of crude oil on the temporal dynamics of phytoplankton and heterotrophic bacteria in Israeli coastal waters. To this end, we added crude oil (500 µm thick layer, with and without additional nutrients; NO₃ and PO₄) to mesocosms (1m³ bags) containing oligotrophic surface coastal water collected near Haifa during summer and winter. Changes in phytoplankton biomass, activity and diversity were monitored daily for 5-6 days. Our results demonstrate that crude oil addition resulted in a pronounced decrease in phytoplankton biomass and production rates, while heterotrophic bacterial production increased significantly. Importantly, a few days post addition we found that the oil-degrading bacteria, Oleibacter sp. and Oleispira sp. appeared in the mesocosms and that the addition of nutrients (along with the crude oil) further increased this trend. This suggests that oil-degrading bacteria may be NO₃ and PO₄ limited in Israeli coastal waters. The results of this study should enable us to establish improved science-based environmental policy with respect to handling crude oil pollution in this region.

Keywords: heterotrophic bacteria, nutrients, mesocosm, oil pollution, oligotrophic, phytoplankton

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Authors: Chittapon Jantararussamee, Malai Taweechotipatr, Udomsri Showpittapornchai, Wisuit Pradidarcheep

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Hepatic fibrosis is characterized by collagen accumulation in hepatic lobules following wound healing process. If lefts untreated, it could progress into hepatic cirrhosis, portal hypertension, and liver failure. Probiotics comprise of lactic acid bacteria which are crucial components of the intestinal microflora and possess many beneficial properties. The objective of this study is to investigate the hepatoprotective effects of probiotic lactic acid bacteria (mixture of Lactobacillus paracasei, Lactobacillus casei, and Lactobacillus confusus at a ratio of 1: 1: 1) on thioacetamide-induced liver fibrotic rats in term of histomorphology study. Twenty-four male Wistar rats were randomly divided into four groups with 6 rats each: (A) control, (B) fibrotic, (C) fibrotic+probiotic, and (D) probiotic. Group (A) received daily oral administration of distilled water. Group (B and C) were induced by intraperitoneal injection of thioacetamide (TAA) (200 mg/kg BW) 3 times per week for consecutive 8 weeks. In probiotic-treated group (C and D), the number of a mixture of the viable microbial cells at 10⁹ CFU/ml was administered orally daily. After sacrifice, liver tissues were collected and processed for routine histological technique and stained with Sirius red. It was found that the fibrotic rats showed hepatic injury marked by area of inflammation, hydropic degeneration of hepatocytes, and accumulation of myofibroblast-like cells. The collagen fibers were substantially accumulated in the hepatic lobules. Moreover, probiotic-treated group significantly reduced the accumulation of collagen in rats treated by TAA. The liver damage was found to be lesser in the probiotic-treated group. It was noted that the liver tissues of control and probiotics groups were shown to be normal. Administration with probiotic lactic acid bacteria could improve the histomorphology in fibrotic liver and be useful for prevention of hepatic disorders.

Keywords: liver fibrosis, probiotics, lactic acid bacteria, thioacetamide

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Authors: Syed Makhdoom Hussain, Muhammad Afzal, Farhat Jabeen, Arshad Javid, Tasneem Hameed

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A feeding trial was conducted with Cirrhinus mrigala fingerlings to study the effects of microbial phytase with graded levels (0, 500, 1000, 1500, and 2000 FTUkg-1) by sunflower meal based diet on growth performance and nutrient digestibility. The chromic oxide was added as an indigestible marker in the diets. Three replicate groups of 15 fish (Average wt 5.98 g fish-1) were fed once a day and feces were collected twice daily. The results of present study showed improved growth and feed performance of Cirrhinus mrigala fingerlings in response to phytase supplementation. Maximum growth performance was obtained by the fish fed on test diet-III having 1000 FTU kg-1 phytase level. Similarly, nutrient digestibility was also significantly increased (p<0.05) by phytase supplementation. Digestibility coefficients for sunflower meal based diet increased 15.76%, 17.70%, and 12.70% for crude protein, crude fat and apparent gross energy as compared to the reference diet, respectively at 1000 FTU kg-1 level. Again, maximum response of nutrient digestibility was recorded at the phytase level of 1000 FTU kg-1 diet. It was concluded that the phytase supplementation to sunflower meal based diet at 1000 FTU kg-1 level is optimum to release adequate chelated nutrients for maximum growth performance of C. mrigala fingerlings. Our results also suggested that phytase supplementation to sunflower meal based diet can help in the development of sustainable aquaculture by reducing the feed cost and nutrient discharge through feces in the aquatic ecosystem.

Keywords: sunflower meal, Cirrhinus mrigala, growth, nutrient digestibility, phytase

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Authors: Assel E. Molzhigitova, Amankeldi K. Sadanov, Elvira T. Ismailova, Kulyash A. Iskandarova, Olga N. Shemshura, Ainur I. Seitbattalova

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Fire blight is a very harmful for commercial apple and pear production quarantine bacterial disease. To date, several different methods have been proposed for disease control, including the use of copperbased preparations and antibiotics, which are not always reliable or effective. The use of bacteria as biocontrol agents is one of the most promising and eco-friendly alternative methods. Bacteria with protective activity against the causal agent of fire blight are often present among the epiphytic microorganisms of the phyllosphere of host plants. Therefore, the main objective of our study was screening of local epiphytic bacteria as possible antagonists against Erwinia amylovora, the causal agent of fire blight. Samples of infected organs of apple and pear trees (shoots, leaves, fruits) were collected from the industrial horticulture areas in various agro-ecological zones of Kazakhstan. Epiphytic microorganisms were isolated by standard and modified methods on specific nutrient media. The primary screening of selected microorganisms under laboratory conditions to determine the ability to suppress the growth of Erwinia amylovora was performed by agar-diffusion-test. Among 142 bacteria isolated from the fire blight host plants, 5 isolates, belonging to the genera Bacillus, Lactobacillus, Pseudomonas, Paenibacillus and Pantoea showed higher antagonistic activity against the pathogen. The diameters of inhibition zone have been depended on the species and ranged from 10 mm to 48 mm. The maximum diameter of inhibition zone (48 mm) was exhibited by B. amyloliquefaciens. Less inhibitory effect was showed by Pantoea agglomerans PA1 (19 mm). The study of inhibitory effect of Lactobacillus species against E. amylovora showed that among 7 isolates tested only one (Lactobacillus plantarum 17M) demonstrated inhibitory zone (30 mm). In summary, this study was devoted to detect the beneficial epiphytic bacteria from plants organs of pear and apple trees due to fire blight control in Kazakhstan. Results obtained from the in vitro experiments showed that the most efficient bacterial isolates are Lactobacillus plantarum 17M, Bacillus amyloliquefaciens MB40, and Pantoea agglomerans PA1. These antagonists are suitable for development as biocontrol agents for fire blight control. Their efficacies will be evaluated additionally, in biological tests under in vitro and field conditions during our further study.

Keywords: antagonists, epiphytic bacteria, Erwinia amylovora, fire blight

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