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Abstract

Bioremediation is based on microorganisms able to use pollutants either as a source of carbon or in co-metabolism, and is a promising strategy in cleaning the environment. Using soil contaminated with petroleum products from an industrial area in Saudi Arabia (Jubail), and after enrichment with the polycyclic aromatic hydrocarbon (PAH) naphthalene, a Methylobacterium radiotolerans strain (N7A0) was isolated that can grow in the presence of naphthalene as the sole source of carbon. M. radiotolerans is known to be resistant to gamma radiation, and this is the first documented report of a strain of this bacterium using a PAH as the sole source of carbon. The commonly reported Pseudomonas aeruginosa (strain N7B1) that biodegrades naphthalene was also identified, and gas chromatography analyses have shown that the biodegradation of naphthalene by M. radiotolerans and P. aeruginosa did follow both the salicylate and phthalate pathways.

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Authors and Affiliations

Alexis Nzila
Assad Thukair
Saravanan Sankara
Basheer Chanbasha
Musa M. Musa
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Abstract

The suitability of low-cost impedance sensors for microbiological purposes and biofilm growth monitoring was evaluated. The sensors with interdigitated electrodes were fabricated in PCB and LTCC technologies. The electrodes were golden (LTCC) or gold-plated (PCB) to provide surface stability. The sensors were used for monitoring growth and degradation of the reference ATCC 15442 Pseudomonas aeruginosa strain biofilm in invitro setting. During the experiment, the impedance spectra of the sensors were measured and analysed using electrical equivalent circuit (EEC) modelling. Additionally, the process of adhesion and growth of bacteria on a sensor’s surface was assessed by means of the optical and SEM microscopy. EEC and SEM microscopic analysis revealed that the gold layer on copper electrodes was not tight, making the PCB sensors susceptible to corrosion while the LTCC sensors had good surface stability. It turned out that the LTCC sensors are suitable for monitoring pseudomonal biofilm and the PCB sensors are good detectors of ongoing stages of biofilm formation.

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Authors and Affiliations

Konrad Chabowski
Adam F. Junka
Tomasz Piasecki
Damian Nowak
Karol Nitsch
Danuta Smutnicka
Marzenna Bartoszewicz
Magdalena Moczała
Patrycja Szymczyk
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Abstract

Petroleum pollution is still one or crucial environmental problems. Bioaugmcntation is a popular technique used in soil remediation. The aim of soil inoculation is acceleration or decomposition processes or improving the degradation efficiency. Effectiveness of bioaugmentation processes depends on the number and activity of microorganisms adapted to pollutant degradation. Enhancement of microorganisms' activity can be reached by the use or plants. Roots or plants excrete organic substances that stimulate microorganisms' growth. Among different species or plants interesting arc papilionaceous plants because of their nitrogen fixation ability in symbiosis with bacteria. The effects or using papilionaceous plants (Trifolium pratense), multiplied autochthonous microorganisms and commercial biopreparation in aged-petroleumpolluted soil were studied. The samples of soil were taken from the refinery in Czechowice-Dziedzice (Poland) and classified as heavily degraded with a C/N-ratio or 100:0.7. Investigations were conducted for 14 weeks. Microbiological analysis included: total bacteria, fungi, Actinomycetes and Pscudonionas counts. Concentration or heavy fractions, TPH (total petroleum hydrocarbons) and PAHs (polycyclic aromatic hydrocarbons) were measured at the start and at the end of the experiment. Presence of pap il ionaceous plant (Trifolium pratensei enhanced the growth or microorganisms, nitrogen concentration and biodegradation processes (removal of 63% or TPH, 44% or heavy fractions, 9% or 4-6 aromatic PAHand 80% of 2-3 aromatic PAH) in polluted soil. An increasing number of Pseudomonas species was observed in samples in which pollution removal was more effective.
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Authors and Affiliations

Ewa Zabłocka-Godlewska
Wioletta Przystaś
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Abstract

Taro leaf blight caused by Phytophthora colocasiae affects plant health and is a major threat to taro culture in Cameroon. Chemical fertilizers used often harm the ecosystem. Plant growth-promoting rhizobacteria (PGPR) are better alternatives that increase plant growth promotion and suppress phytopathogens. In the present study, a total of 67 fluorescent Pseudomonas spp. was characterized by 17.91, 5.97, and 4.47% populations of P. fluorescens, P. chlororaphis, and P. putida, respectively, among the most represented. More than 36% of bacteria showed antagonistic potential through the production of both diffusible and volatile compounds. Some of them (03) exhibited antagonistic activity in dual culture against P. colocasiae with a diameter greater than 13 mm. These rhizobacteria produced a significant amount of siderophore, IAA, SA, HCN, protease, lipases, and cellulases. For the pot experiment, treatment by Pseudomonas significantly increased the enzymatic activity involved in the resistance of taro, such as peroxidase (PO), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL). The two antagonists also increased plant growth parameters of taro such as chlorophyll, plant height, shoot length, total leaf surface, fresh root biomass, and fresh leaf biomass. These findings showed that fluorescent Pseudomonas have an intriguing and undeniable potential in the fight against P. colocasiae, which could lead to the development of a biopesticide in the future.
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Authors and Affiliations

Samuel Arsène Ntyam Mendo
1
ORCID: ORCID
Dorice Nguelo Dzumafo
2
Laure Brigitte Kouitcheu Mabeku
3
Severin Tchameni Nguemezi
2
ORCID: ORCID
Lambert Sameza Modeste
2
Rosalie Anne Ngono Ngane
2

  1. Department of Biological Sciences, Higher Teacher Training College, University of Yaounde 1, Yaounde, Cameroon
  2. Department of Biochemistry, Faculty of Sciences, University of Douala, Douala, Cameroon
  3. Department of Microbiology, Faculty of Science, University of Yaoundé 1, Yaounde, Cameroon
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Abstract

A total of 94 pectolytic and 60 nonpectolytic Pseudomonas isolates were obtained from 250 samples of rotted vegetable specimens representing various economically important vegetables. The isolates were identified on the basis of standard biochemical tests. Pseudomonas fluorescens biovar V and II and Pseudomonas putida were the most abundant species among pectolytic isolates and Pseudomonas fluorescens biovar I among nonpectolytic ones. Only 3 Pseudomonas viridiflava isolates were identified and all of them were obtained from potato. Isolates of pectolytic phenotype were scattered among nonpectolytic ones irrespective of their taxonomical status. Isolates identified biochemically, as Pseudomonas marginalis were also present in nonpectolytic group. PCR method is unsuitable for identification and differentiation of bacteria belonging to pectolytic fluorescens Pseudomonas group due to great diversity of species. However, the results of PCR amplification of the genes encoding pectate lyase suggest that genes responsible for production of this enzyme may also be present in isolates of nonpectolytic phenotype.
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Authors and Affiliations

Lena Żołbowska
Henryk Pospieszny
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Abstract

Fluorescent Pseudomonas (FP) is a major group of plant growth promoting rhizobacteria and a well-known synthesizer of siderophores, which imparts a selective advantage on rhizosphere competence and their biocontrol traits. The present study was aimed at examining the factors affecting the production of siderophores and their potential biocontrol traits. Sixteen FP isolates were shortlisted based on their siderophore-producing ability in chrome azural S medium. The isolates were checked for variations in siderophore production under varying incubation times, temperatures, pH, iron (Fe3+) concentrations and mutagens. In addition, the iron binding affinity of siderophores, mycelial inhibition assay and plant growth promotion traits were assessed. Results showed that the siderophore production was highly influenced by the time of incubation, changes in pH, temperature and iron concentration. Chemical characterization showed that the produced siderophores were hydroxamates. Maximum siderophore production was observed at pH 7 whereas UV and EtBr exposure invariably suppressed siderophore production drastically in all isolates. All FPs from maize rhizosphere showed excellent siderophore production which could be due to the competence in strategy-II of the plant rhizosphere and significant growth inhibition on Fusarium oxysporum. Our results suggest the inclination of siderophores to iron, in terms of various criteria affecting production and the possible role of environmental mutations that affect the natural iron harvesting mechanism.
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Authors and Affiliations

Deori Monali
Jayamohan Nellickal Subramanyan
Kumudini Belur Satyan
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Abstract

In the present study a severe outbreak of hemorrhagic pneumonia (HP) in neonatal minks concomitant with Leismania infantum (L. infantum) detection is reported. The outbreak took place on a Greek mink farm and affected 1,362 mink kits, with 524 dying. Macroscopic lesions of 14 necropsied affected kits were confined to the respiratory system with dark red, consolidated lung lobes and to the small intestine with severe, acute, hemorrhagic and necrotic enteritis. Microscopic examination of lung sections revealed severe hemorrhagic pyogranulomatous pneumonia. Bacteria were obtained in pure culture from the lungs of all necropsied animals and were confirmed as Pseudomonas aeruginosa (P. aeruginosa). Three out of 14 (21.4%) animals were positive for the presence of L. infantum DNA. The outbreak was attributed to the infection of minks with P. aeruginosa, possibly as a consequence of being immuno-suppressed by L. infantum. Further research is necessary, especially on the pathogenesis of P. aeruginosa/L. infantum co-infection and the implications of this interaction on HP disease outcome.
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Authors and Affiliations

G. Filioussis
E. Petridou
D. Papadopoulos
E. Karavanis
E. Morgan
C. Billinis
E. Papadopoulos
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Abstract

Excessive use of chemical fertilizers, in agriculture, has negative impacts on water, soil and affects the environment and health. In recent decades, researchers have been interested in the natural benefits of natural microorganisms and how they could be a good alternative to the use of chemical fertilizers. The aim of this study was to investigate the effect of soil inoculation with strains of mycorrhizae and beneficial bacteria on soil properties and productivity of table grapes. Field trials were conducted on a commercial table grape production farm ( Vitis vinifiera cv. Mousca), located in northeastern Morocco. Twelve-yearold plants were used. Control plants were not inoculated (T1). The prototype plants were inoculated with 1.2 × 104 of Glomus iranicum var. tenuihypharum/100 g (T2), a mixture of 1/2 concentration of Glomus iranicum var. tenuihypharum and 1/2 concentration of Pseudomonas putida (T3) and 1 × 108 CFU ∙ g–1 of Pseudomonas putida (T4). The inoculations were realized twice; the first inoculation was completed on July 19, 2019 while the second inoculation on February 21, 2020. Soil analyses were carried out, both physicochemical (pH, electrical conductivity (EC), salinity, % of dry matter) and microbiological properties (total flora, fungi and actinobacteria). Plant growth (length of the plant, number and diameter of sticks, number of clusters per tree, number of nodes per stick, distance between nodes and bud burst), yield and fruit quality (number of berries per cluster, cluster weight, cluster length and width, pH, Brix degrees, acidity, EC and % dry matter) were measured. Results showed slight trends regarding the effects of treatments on the physicochemical and microbiological properties of the soil, plant growth and fruit quality. The number of clusters was significantly higher in Glomus (T2) Pseudomonas (T4) and Glomus than in control treatments.
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Authors and Affiliations

Salah Ed-dine Samri
1
ORCID: ORCID
Kamal Aberkani
1
ORCID: ORCID
Mourad Said
1
Khadija Haboubi
2
ORCID: ORCID
Hassan Ghazal
3
ORCID: ORCID

  1. Biology and Geology, Plolydisciplinary Faculty of Nador, University Mohammed Fisrt, Selonane, Morocco
  2. Environment, National School of Applied Sciences, University Abdelmalek Essaadi, Al Hoceima, Morocco
  3. Bioinformatics, National Center for Scientific and Technical Research, Rabat, Morocco
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Abstract

In the present work, bacteria of the Pseudomonas genus native to the Ecuadorian Amazon with the capacity to treat contaminated water and soils were selected. For this purpose, 20 soil samples from Amazon region with evidence of contamination were analysed. For identification, each sample was assigned a code according to the sampling area: Joya de los Sachas (S), Minga (M) and Siete de Julio-Shushufindi (SH). The cultures were performed in the combination of Bushnell Hass (BH) + Luria Bertani (LB) and Müeller-Hinton (MH) + Brucella agar (BA) media, all with the addition of diesel to verify their efficacy in the growth of bacteria capable of surviving in contaminated media. The combination with ideal results was that of BH + LB, by means of Gram-staining it was determined that 19 of the samples had interest microorganisms. To characterize the isolates at the species level, biochemical tests of: catalase, citrate, glucose, hemolytic activity and urease were applied, which allowed to confirm the existence of the Pseudomonas of interest. The results indicated that P. stutzeri (in samples S1 and M1), P. aeruginosa (in SH2 and SH5) and P. putida (in S7, S8, S10 and SH4) obtaining a total of 8 isolates (40%) of interest from the initial 19. With the results obtained from this work, an optimal culture method was standardized for the selection of bacteria with potential for treating contaminated soils and water.
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Authors and Affiliations

Henry Joel Escudero-López
1
ORCID: ORCID
Karla Anabel Serrano-Carrillo
2
ORCID: ORCID
Carlos Rodrigo Jácome-Pilco
1
ORCID: ORCID
Herminia del Rosario Sanaguano-Salguero
1
ORCID: ORCID
Isidro Favian Bayas-Morejón
1
ORCID: ORCID

  1. State University of Bolívar, Faculty of Agricultural Sciences, Agroindustry Career, Biotechnological Research and Development Center, Km 3 1/2 sector Alpachaca, CP: 020150, Guaranda, Ecuador
  2. San Pedro Educational Unit, Department of Education, Guaranda, Ecuador
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Abstract

Carbonic anhydrase is an important enzyme that can play a significant role in the processes of lowering carbon dioxide concentration in the atmosphere. The aim of the work was to investigate the extracellular carbonic anhydrase (CA) production by the bacteria Pseudomonas fragi. In the research, we focused on the evaluation of the phase of bacterial growth correlated with carbonic anhydrase production and on the evaluation of induction of CA production by calcium carbonate concentration in the nutrient medium. Presented data indicated that calcium carbonate can serve as the only carbon source for Pseudomonas fragi, inducing carbonic anhydrase secretion to culture broth. The enzyme was produced mainly in the adaptation growth phase reaching the maximal activity at the end of this phase or at the beginning of the growth phase. The maximal enzyme activity detected in all batches was at a similar level. The enzyme activity was constant but lower in the exponential phase growth. Therefore, the enzyme production is not growth-dependent, but it is correlated with bacteria adaptation to cultivation conditions.
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Authors and Affiliations

Andrzej Tietz
1
Małgorzata M. Jaworska
1
ORCID: ORCID

  1. Warsaw University of Technology, Faulty of Chemical and Process Engineering, ul. Warynskiego 1, 00-645 Warsaw, Poland
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Abstract

In a dairy cow herd consisted of herd of 200 lactating Holstein-Friesian cows and heifers, clinical signs of mastitis in 40 out of 170 animals were observed. Treatments with antibiotics were proved ineffective. Milk bacterial cultures from 15 affected animals revealed Pseudomonas aeruginosa. An autogenous vaccine was administered subcutaneously, twice in a month period, to all adults. Cases of clinical mastitis declined significantly (p<0.0001) during next 3 months.
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Authors and Affiliations

E.J. Petridou
1
I.A. Fragkou
2
S.Q. Lafi
3
N.D. Giadinis
4

  1. Laboratory of Microbiology and Infectious Diseases, Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, Greece
  2. Veterinary Service, Ministry of Rural Development and Food, Athens, Greece
  3. Department of Pathology and Animal Health, Faculty of Veterinary Medicine, Jordan University of Science and Technology, P.O. Box 3030, Irbid, Jordan
  4. Clinic of Farm Animals, Faculty of Veterinary Medicine, Aristotle University of Thessaloniki, Greece
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Abstract

Influence of Fusarium oxysporum f. sp. cubense (E.F. Smith) Snyder and Hansen on 2,4-diacetylphloroglucinol (DAPG) production in the rhizosphere of banana cultivar Rasthali by Pseudomonas fluorescens was investigated. The purified extracts of Pfm strain of P. fluorescens isolated from banana rhizosphere inhibited the growth and spore germination of F. oxysporum f. sp. cubense under laboratory conditions. DAPG extracted from the cultures of the strain was observed as distinct spots in thin layer chromatographic plates at Rf value of 0.88. The extracts of soil inoculated with P. fluorescens and challenge inoculated with F. oxysporum f. sp. cubense eluted at retention time ranges from 20.00 min to 21.30 min. The quantity of DPAG production was less in the extracts of soil inoculated with P. fluorescens and challenge inoculated with F. oxysporum f. sp. cubense as compared to P. fluorescens alone inoculated soil. The talc formulation of Pfm strain also reduced vascular discolouration due to the pathogen in banana plants when inoculated at 15 g/plant.

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Authors and Affiliations

Thangavelu Saravanan
Mari Muthusamy
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Abstract

Studies were carried out in the summer seasons of l 995-1999 (from June to September) on the quantitative and qualitative composition of Enterobacteriaceae bacteria (including Escherichia coli and Salmonella sp.), and potentially pathogenic bacteria Aeromonas hydrophila, Pseudomonas aeruginosa and Staphylococcus sp. in the water of 8 bathing sites of the Lake Wigry. Aeromonas hydrophila occurred in the all samples of studied water and was the most numerous in water sampled from sites of increased trophic levels. Irrespective of the site and time of sampling Pseudomonas aeruginosa was rarely isolated. In the total of 160 samples of water analysed Salmonella sp.. Escherichia coli and Staphyloccocus aureus were determined in 32 (22.6%), 68 (42.4%) and 90 (55%) samples, respectively. Pathogenic bacteria of the genus Salmonella sp. and potentially pathogenic Staphylococcus aureus and Aeromonas hydrophila identified in the analysed offshore waters (also when Escherichia coli were absent) suggest that the use of the Lake Wigry waters for swimming, falling into account faecal bacterial counts, may not be sufficient to fully reflect safety conditions for bathers. The results of the research suggest that the evaluation of the Lake Wigry surface waters for recreational use should include the frequency of the occurrence of Salmonella sp., Staphylococcus aureus, Aeromonas hydrophila and Pseudomonas aeruginosa. These three species, which arc not directly linked to faecal contamination, can cause various diseases of the skin, nasal and oral cavities, eyes, internal car and other problems in people swimming in contaminated water.
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Authors and Affiliations

Ewa Korzeniewska
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Abstract

In the last decade, atmospheric plasma has been used to treating bacterial infections in humans due to its bactericidal effects; however, its efficacy in dogs is unclear. This study evaluated the in vitro bactericidal efficacy of atmospheric plasma on Staphylococcus pseudinter- medius and Pseudomonas aeruginosa, two of the most important bacterial agents isolated from canine pyodermas. Three isolates each of S. pseudintermedius and P. aeruginosa obtained from dogs with pyoderma were subjected to atmospheric plasma. The isolates from the control group were not exposed to plasma, while those from the treatment groups were exposed to plasma for 15 (7.5 J/cm2), 30 (15 J/cm2), 60 (30 J/cm2), or 90 (45 J/cm2) seconds. After each treatment, a reduction in colony formation was observed. Bacterial viability was evaluated using the LIVE/ DEAD® BacLight™ Bacterial Viability Kit. The antibacterial effects were evaluated with Image J software and significance was assessed statistically in comparison to the control group. The bactericidal effect of atmospheric plasma against both bacteria increased significantly in a time-dependent manner. These results demonstrate the bactericidal capacity of atmospheric plasma, and suggest that it could serve as an alternative treatment method for canine pyoderma. Further studies are needed to evaluate the safety and efficacy of atmospheric plasma in dogs.

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Authors and Affiliations

S. Bae
D. Lim
D. Kim
J. Jeon
T. Oh
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Abstract

Pseudomonas spp. are a psychrotrophic species associated with milk spoilage caused by its enzymatic activities. The aim of this study was to identify Pseudomonas spp. in raw cow’s milk and to investigate their associated enzymatic properties and the ability to produce pyoverdine pigment. For microbiological analysis, 2 ml of milk samples was taken in sterile sample boxes. Milk sampling was carried out according to the principles of STN EN ISO 707. By identification using the PCR method, of n=320 samples of raw cow milk a total of 73 isolates of Pseudomonas spp. were identified as P. putida (34.25%); P. fragi (13.70%); P. lundensis (9.59%) and Pseudomonas spp. (42.47%). Proteolytic activity determined at a temperature of 7°C was positive from n=20 selected isolates of Pseudomonas spp. (60%) isolates and a temperature of 25°C was positively detected (85%). Lipolytic activity determined at a temperature of 7°C was confirmed in 35% of isolates and a temperature of 25°C it was confirmed in 70% isolates. Pyoverdine pigment production was detected in 65% of isolates. The results reveal the enzymatic activity of Pseudomonas spp. present in raw cow’s milk and its spoilage potential at different temperatures in relation to pigment production.
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Authors and Affiliations

M. Kováčová
1
J. Výrostková
1
I. Regecová
1
E. Dudriková
1
J. Zahumenská
1
S. Marcinčák
1

  1. Department of Food Hygiene Technology and Safety, University of Veterinary Medicine and Pharmacy in Košice, Komenského 73, 041 81 Košice, Slovakia
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Abstract

The purpose of the presented studies was to determine the species composition of the fungi occurring on the underground parts of potato at an thesis and to establish the qualitative and quantitative composition ofmicroorganisms living in the soil environment of this plant. Besides, the studies aimed at finding antagonistic microorganisms inhibiting potato infection by soil-borne phytopathogens. The mycological analysis showed that the symptoms of necrosis on the roots and the stem base of potato were caused by Alternaria alternata, Fusarium spp., Colletotrichum coccodes and Rhizoctonia so/ani. The microbiological analysis of the potato rhizosphere gave twice as many bacteria and fungi as from the non-rhizosphere soil. The dominating pathogenic fungi in the examined soil samples were Fusarium spp., A. alternata and R. so/ani. Three times as many antagonistic bacteria Pseudomonas spp. and Bacillus spp. and more than twice as many antagonistic fungi (Penicillium spp., Trichoderma spp., Gliocladium spp.) were obtained from the potato rhizosphere as compared to the non-rhizosphere soil.
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Authors and Affiliations

Danuta Pięta
Elżbieta Patkowska

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