PVA-Based Films with Strontium Titanate Nanoparticles Dedicated to Wound Dressing Application.

Bioactive materials may be applied in tissue regeneration, and an example of such materials are wound dressings, which are used to accelerate skin healing, especially after trauma. Here, we proposed a novel dressing enriched by a bioactive component. The aim of our study was to prepare and characterize poly(vinyl alcohol) films modified with strontium titanate nanoparticles. The physicochemical properties of films were studied, such as surface free energy and surface roughness, as well as the mechanical properties of materials. Moreover, different biological studies were carried out, like in vitro hemo- and cyto-compatibility, biocidal activity, and anti-biofilm formation. Also, the degradation of the materials' utilization possibilities and enzymatic activity in compost were checked. The decrease of surface free energy, increase of roughness, and improvement of mechanical strength were found after the addition of nanoparticles. All developed films were cyto-compatible, and did not induce a hemolytic effect on the human erythrocytes. The PVA films containing the highest concentration of STO (20%) reduced the proliferation of Eschericha coli, Pseudomonas aeruginosa, and Staphylococcus aureus significantly. Also, all films were characterized by surface anti-biofilm activity, as they significantly lowered the bacterial biofilm abundance and its dehydrogenase activity. The films were degraded by the compost microorganism. However, PVA with the addition of 20%STO was more difficult to degrade. Based on our results, for wound dressing application, we suggest using bioactive films based on PVA + 20%STO, as they were characterized by high antibacterial properties, favorable physicochemical characteristics, and good biocompatibility with human cells.

Characterization of bacterial biofilms developed on the biodegradable polylactide and polycaprolactone polymers containing birch tar in an aquatic environment.

Birch tar was added to polylactide (PLA) and polycaprolactone (PCL) to create films with antimicrobial properties. After incubating the films for seven days in lake water, the diversity of bacterial communities developed on the surfaces of PCL and PLA with embedded birch tar (1 %, 5 %, and 10 %, w/w) was assessed with amplicon sequencing of the 16S rRNA gene on a MiSeq platform (Illumina). Notably, Aquabacterium and Caulobacter were more abundant at the surface of PCL compared to PLA (13.4 % vs 0.2 %, p < 0.001 and 9.5 % vs 0.2 %, p < 0.001, respectively) while Hydrogenophaga was significantly more abundant at the surface of PLA compared to PCL (6.1 % vs 1.8 %, p < 0.01). Overall, lower birch tar concentrations (1 % and 5 % on both polymers) stimulated bacterial diversity in biofilms compared to the control. The number of reeds assigned to Flavobacterium and Aquabacterium showed a rising trend with the increase of birch tar concentration on the surface of both polymers.

Carboxymethyl cellulose based films enriched with polysaccharides from mulberry leaves (Morus alba L.) as new biodegradable packaging material.

The objective of this study was to determine the properties of a new active packaging film developed by the addition of mulberry leaves polysaccharides (MLP) into carboxymethyl cellulose (CMC). Biodegradable CMC-MLP films were fabricated by casting method with various concentrations of MLP (1, 5 and 10 % w/w). The addition of MLP into the CMC matrix resulted increased thickness (0.126 to 0.163 mm) and roughness of the films. Also, the decline in moisture content from 27.91 to 14.12 %, water vapor permeability from 8.95 to 5.21 × 10-10 g-1 s-1 Pa-1, and a swelling degree from 59.11 to 37.45 % were observed. With the increasing concentration of MLP, the mechanical properties of the films were improved and higher dispersion of UV light were noted. Fourier transform - infrared spectroscopy (FT-IR) and X-ray diffraction revealed good inter-molecular interaction between CMC matrix and MLP. The prepared films showed excellent thermal stability, antioxidant and antibacterial properties as well as susceptibility to biodegradation in the soil environment. Moreover, it was proved that the films have ability to retard oil oxidation. Overall, it was concluded that CMC-MLP films constitute a promising biomaterial that may be applied as active food packaging.

The effect of seed bacterization with Bacillus paralicheniformis 2R5 on bacterial and fungal communities in the canola rhizosphere.

Bacillus sp. is one of the best-studied plant growth-promoting rhizobacteria (PGPR). However, more detailed studies targeting its effect on the rhizosphere microbial community are required for improving management practices regarding its commercial application in the field. Our earlier study showed that PGPR Bacillus paralicheniformis 2R5 stimulated canola growth. Hence, this study aimed to assess the time-course impact of B. paralicheniformis 2R5 on bacterial and fungal community structure and diversity. The results showed that inoculation with B. paralicheniformis 2R5 initially significantly decreased the observed bacterial richness compared to the control, while after 44 days of treatment this alpha diversity metrics increased. A linear discriminant analysis effect size showed that B. paralicheniformis 2R5 altered the soil bacterial and fungal community structure by increasing the abundance of plants' beneficial microorganisms such as Nitrospira, Ramlibacter, Sphingomonas, Massilia, Terrimonas as well as Solicoccozyma, Schizothecium, Cyphellophora, Fusicolla, Humicola. B. paralicheniformis 2R5 seems to be a promising alternative to chemical pesticides and can be considered for practical application in the field. Its ability to alter the rhizosphere microbiome by increasing the diversity and composition of bacterial communities and increasing plants' beneficial groups of fungi, appears to be important in terms of improving canola development. However, further studies on these increased microbial taxa are necessary to confirm their function in promoting canola growth.

Effect of birch tar embedded in polylactide on its biodegradation.

The plasticized film was made of polylactide and birch tar, which was used in a concentration of 1, 5 and 10 % by weight. Tar was added to the polymer to obtain materials with antimicrobial properties. The main purpose of this work is to characterize and biodegradation of this film after the end of its use. Therefore, the following analyzes were performed: enzymatic activity of microorganisms in the presence of polylactide (PLA) film containing birch tar (BT), biodegradation process in compost, barrier changes and structural properties of the film before and after biodegradation and bioaugmentation. Biological oxygen demand BOD21, water vapor permeability (Pv), oxygen permeability (Po), scanning electron microscopy (SEM) and enzymatic activity of microorganisms were assessed. Microorganism strains Bacillus toyonensis AK2 and Bacillus albus AK3 were isolated and identified, which constituted an effective consortium increasing the susceptibility of polylactide polymer material with tar to biodegradation in compost. Analyses with the use of the above-mentioned strains had an impact on the change of physicochemical properties, e.g. the presence of biofilm on the surface of the analyzed films and the reduction of the barrier properties of the film, which translates into the recorded susceptibility to biodegradation of these materials. The analyzed films can be used in the packaging industry, and after use, subjected to intentional biodegradation processes, including bioaugmentation.

Physiological properties and genomic insights into the plant growth-promoting rhizobacterium Brevibacillus laterosporus K75 isolated from maize rhizosphere.

BACKGROUND: When looking for a safer alternative to pesticides that are potentially harmful to living organisms, one of the directions worth looking at are plant growth-promoting rhizobacteria. The purpose of the research was a comprehensive characterization of Brevibacillus laterosporus K75, a strain isolated from maize rhizosphere. Many studies have proved B. laterosporus to be a biocontrol agent; however, little is known about B. laterosporus as a plant growth-promoting rhizobacterium. RESULTS: Ninety strains were screened for plant growth-promoting activities. Four strains with the best plant growth-promoting traits (Rhodococcus qingshengii K8, Bacillus subtilis subsp. stercoris K73, Brevibacillus laterosporus K75, and Brevibacillus laterosporus K89) were used to research their effect on maize growth. Under sterile conditions, B. laterosporus K75 showed the best stimulatory effect, significantly improving the weight of roots, shoots and leaves, and considerably increasing content of chlorophyll. In unsterilized soil, B. laterosporus K75 significantly improved length of roots and weight of leaves compared to the K73, K89, and untreated control. Moreover, B. laterosporus K75 significantly increased specific leaf area compared to the untreated control and to other inoculant treatments. The genome of B. laterosporus K75 was compared to the recently published B. laterosporus MG64. Genome-mining displayed differences in identified plant growth-promoting genes and biosynthetic gene clusters of secondary metabolites. The B. laterosporus K75 genome possessed additional genes involved in indole-3-acetic acid production and phosphate solubilization that could be attributed to its ability to enhance maize growth. CONCLUSION: Our study demonstrated that B. laterosporus K75 is a promising candidate for use in inoculant formulation, effectively facilitating maize growth. © 2022 Society of Chemical Industry.

Plant growth-promoting rhizobacteria: Peribacillus frigoritolerans 2RO30 and Pseudomonas sivasensis 2RO45 for their effect on canola growth under controlled as well as natural conditions.

Even though canola is one of the most important industrial crops worldwide, it has high nutrient requirements and is susceptible to pests and diseases. Therefore, natural methods are sought to support the development of these plants. One of those methods could be a plant growth-promoting rhizobacteria (PGPR) that have a beneficial effect on plant development. The aim of this study was a genomic comparison of two PGPR strains chosen based on their effect on canola growth: Peribacillus frigoritolerans 2RO30, which stimulated canola growth only in sterile conditions, and Pseudomonas sivasensis 2RO45, which promoted canola growth in both sterile and non-sterile conditions. First of all, six bacterial strains: RO33 (Pseudomonas sp.), RO37 (Pseudomonas poae), RO45 (Pseudomonas kairouanensis), 2RO30 (Peribacillus frigoritolerans), 2RO45 (Pseudomonas sivasensis), and 3RO30 (Pseudomonas migulae), demonstrating best PGP traits in vitro, were studied for their stimulating effect on canola growth under sterile conditions. P. frigoritolerans 2RO30 and P. sivasensis 2RO45 showed the best promoting effect, significantly improving chlorophyll content index (CCI) and roots length compared to the non-inoculated control and to other inoculated seedlings. Under non-sterile conditions, only P. sivasensis 2RO45 promoted the canola growth, significantly increasing CCI compared to the untreated control and to other inoculants. Genome comparison revealed that the genome of P. sivasensis 2RO45 was enriched with additional genes responsible for ACC deaminase (acdA), IAA (trpF, trpG), and siderophores production (fbpA, mbtH, and acrB) compared to 2RO30. Moreover, P. sivasensis 2RO45 showed antifungal effect against all the tested phytopathogens and harbored six more biosynthetic gene clusters (BGC), namely, syringomycin, pyoverdin, viscosin, arylpolyene, lankacidin C, and enterobactin, than P. frigoritolerans 2RO30. These BGCs are well known as antifungal agents; therefore, it can be assumed that these BGCs were responsible for the antifungal activity of P. sivasensis 2RO45 against all plant pathogens. This study is the first report describing P. sivasensis 2RO45 as a canola growth promoter, both under controlled and natural conditions, thus suggesting its application in improving canola yield, by improving nutrient availability, enhancing stress tolerance, and reducing environmental impact of farming practices.

Detection, characterization, and antimicrobial susceptibility of Yersinia enterocolitica in different types of wastewater in the Czech Republic.

AIMS: The purpose of the study was to evaluate the occurrence of Yersinia enterocolitica in different types of wastewater and to characterize the isolates by biotyping, serotyping, and antimicrobial susceptibility testing. In addition, cultivation protocols were evaluated. METHODS AND RESULTS: The occurrence of Y. enterocolitica was determined in treated and untreated municipal wastewater, as well as in hospital, slaughterhouse, and cowshed wastewater. Y. enterocolitica was detected in 84.1% of the wastewater samples, while the main sources were untreated municipal and slaughterhouse wastewater. In contrast, the lowest incidence was found in hospital wastewater. An exclusive occurrence of biotype 1A (98.3%) was detected. Pathogenic bio-serotypes 4/O:3 and 3/O:3 were isolated only from slaughterhouse wastewater. The highest resistance rates were observed for ampicillin (92.5%) and amoxicillin-clavulanic acid (36.8%). CONCLUSIONS: Y. enterocolitica was commonly detected in wastewater, although the prevalence varied depending on the origin of the wastewater. No single cultivation protocol was able to recover Y. enterocolitica isolates from such a complex matrix as wastewater. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study provided data that can contribute to the evaluation of wastewater as a source of Y. enterocolitica and to understanding the threat of wastewater isolates to human health.

Biodegradability of Novel Polylactide and Polycaprolactone Materials with Bacteriostatic Properties Due to Embedded Birch Tar in Different Environments.

The microbial biodegradation of new PLA and PCL materials containing birch tar (1-10% v/v) was investigated. Product of dry distillation of birch bark (Betula pendula Roth) was added to polymeric materials to obtain films with antimicrobial properties. The subject of the study was the course of enzymatic degradation of a biodegradable polymer with antibacterial properties. The results show that the type of the material, tar concentration, and the environment influenced the hydrolytic activity of potential biofilm degraders. In the presence of PCL films, the enzyme activities were higher (except for α-D-glucosidase) compared to PLA films. The highest concentration of birch tar (10% v/v) decreased the activity of hydrolases produced by microorganisms to the most significant extent; however, SEM analysis showed the presence of a biofilm even on plastics with the highest tar content. Based on the results of the biological oxygen demand (BOD), the new materials can be classified as biodegradable but, the biodegradation process was less efficient when compared to plastics without the addition of birch tar.

The role of microorganisms in biodegradation of chitosan/tannic acid materials.

High utilization of thermoplastic polymers with low degradation rates as packaging materials generates a large amount of waste. Therefore, it should be replaced by natural polymers that can be degraded by microorganisms. In this paper, chitosan (CTS)/tannic acid (TA) materials in the weight ratios of 80CTS/20TA and 50CTS/50TA were prepared as potential packaging materials. The results showed that these materials were similarly degraded in soil and compost. However, in comparison to 50CTS/50TA, 80CTS/20TA was slightly better degraded in soil. After 14 days of biodegradation, the chemical structure of materials was changed resulting from adhesion of the microorganisms. The smallest changes were observed on 80CTS/20TA film. Bacterial species were collected and identified from materials after the degradation process. Microorganisms with the highest hydrolytic activity were chosen for the degradation study. Biodegradation and hydrolytic activity were observed only in a few strains, which indicate difficulties in material degradation. Soil bacteria degraded the films better than bacteria isolated from the compost. This study showed also that consortia of bacteria added to soil and compost had a positive effect on the biodegradation of the tested materials and increased the biodegradation of these materials in the studied environments.

Salino-alkaline lime of anthropogenic origin a reservoir of diverse microbial communities.

This paper presents study on the microbiome of a unique extreme environment - saline and alkaline lime, a by-product of soda ash and table salt production in Janikowo, central Poland. High-throughput 16S rDNA amplicon sequencing was used to reveal the structure of bacterial and archaeal communities in the lime samples, taken from repository ponds differing in salinity (2.3-25.5% NaCl). Surprisingly abundant and diverse bacterial communities were discovered in this extreme environment. The most important geochemical drivers of the observed microbial diversity were salinity, calcium ions, nutrients, and water content. The bacterial and archaeal communities in saline, alkaline lime were similar to those found in natural haloalkaline environments. Although the archaeal contribution to the whole microbial community was lower than 4%, the four archaeal genera Natronomonas, Halorubrum, Halobellus, and Halapricum constituted the core microbiome of saline, alkaline lime - a set of OTUs (> 0.1% of total archaeal relative abundance) present in all samples under study. The high proportion of novel, unclassified archaeal and bacterial sequences (not identified at 97% similarity level) in the 16S rRNA gene libraries indicated that potentially new genera, especially within the class of Thermoplasmata inhabit this unique environment.

Alkaline and Halophilic Protease Production by Bacillus luteus H11 and Its Potential Industrial Applications.

This paper presents the results of the study on the production of protease by Bacillus luteus H11 isolated from an alkaline soda lime. B. luteus H11 was identified as an alkalohalophilic bacterium, and its extracellular serine endoprotease also showed an extreme alkali- and halotolerance. It was remarkably stable in the presence of NaCl up to 5 M. The enzyme was active in a broad range of pH values and temperatures, with an optimum pH of 10.5 and a temperature of 45 °C. It had a molecular mass of about 37 kDa and showed activity against azocasein and a synthetic substrate for the subtilisin-like protease, N-succinyl-l-phenylalanine-p-nitroanilide. The halo-alkaline protease produced by B. luteus H11 seems to be significant from an industrial perspective because of its tolerance towards high salinity and alkalinity as well as its stability against some organic solvents, surfactants and oxidants. These properties make the protease suitable for applications in food, detergent and pharmaceutical industries, and also in environmental bioremediation.

Chemical composition, antimicromicrobial activity and insecticidal activity against the lesser mealworm Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) of Origanum vulgare L. ssp. hirtum (Link) and Artemisia dracunculus L. essential oils.

BACKGROUND: Essential oils (EOs) from Artemisia dracunculus L. and Origanum vulgare L. ssp. hirtum were obtained and the qualitative and quantitative chemical composition of the extracts was investigated. The insecticidal activity of EOs against the larval stages of Alphitobius diaperinus (Panzer) was studied. Moreover, the antimicrobial activity of these oils against pathogens transmitted by this pest was also investigated. RESULTS: The obtained results indicate the possibility of using Greek oregano EO with a high content of carvacrol as a feed additive in poultry nutrition. The use of the Greek oregano oil at 1% (w/w) dose showed stronger reduction of body weight gain of stage IV larvae. Their body mass was only 10.92% of the control. Moreover, EOs from O. vulgare strongly inhibited the growth of tested bacterial strains as well as Candida albicans. CONCLUSION: Greek oregano EO may be a good alternative to antibiotic growth promoters and coccidiostats whose use in feeding farm animals has been prohibited since January 2006 under European Union directives. The introduction of O. vulgare L. ssp. hirtum EO into the premises of farm and poultry houses may help to improve sanitary conditions and control of the lesser mealworm inhabiting these buildings. © 2017 Society of Chemical Industry.

Purification, characterization, and gene cloning of a chitinase from Stenotrophomonas maltophilia N4.

The Stenotrophomonas maltophilia synthesises high-activity chitinase in response to chitin or chitosan induction. The enzyme was purified 8.5 fold and subjected to characterisation. The optimum hydrolysis conditions for this enzyme when using colloidal chitin as substrate were pH 5.6 and temperature of 45 °C. The enzyme demonstrated high thermal stability at 45 °C within 2 h. The studied chitinase exhibited high activity towards colloidal chitin, glycol chitin and chitosan, while it did not hydrolyse glycosidic bonds in carboxymethylcellulose. The enzyme exhibited the highest activity, equalling 90 U/ml, towards Nitrophenyl ß-D-N,N',N"-triacetylchitotriose and activity of 37 U/ml towards 4-Nitrophenyl N,N'-diacetyl-ß-D-chitobioside. The K(m) value in the presence of the two former substrates was:1.2 and 3.9 mM, respectively, which classifies the studied enzyme as an endochitinase. Cysteine and 2-mercaptoethanol stimulated to a small degree the activity of the chitinase which may indicate the involvement of cysteine residues in the catalysis mechanism. The full length of the nucleotide sequence of this chitinase gene is 2106 bp, which amounts to 702 amino acids.

The role of open-air inhalatoria in the air quality improvement in spa towns.

OBJECTIVES: The present study was aimed at evaluating microbiological contamination of air in Ciechocinek and Inowroclaw - Polish lowland spa towns. Additionally, the impact of open-air inhalatoria on the quality of air was evaluated. MATERIAL AND METHODS: Air samples were collected seasonally in the urban areas, in the recreation areas and in the vicinity of inhalatoria in both towns using impaction. The numbers of mesophilic bacteria, staphylococci, hemolytic bacteria and actinomycetes were determined on media according to the Polish Standard PN-86/Z-04111/02. The number of moulds was determined on media according to the Polish Standard PN-86/Z-04111/03. RESULTS: While the highest numbers of microorganisms were noted at the sites located in the urban areas, the lowest numbers were noted in the vicinity of the open-air inhalatoria. In all the investigated air samples the values of bioaerosol concentrations were below the recommended TLVs (≤ 5000 CFU×m(-3) for both bacteria and fungi in outdoor environments). Location of the sampling site was invariably a decisive factor in determining the number of microorganisms in the air. CONCLUSIONS: The aerosol which is formed in the open-air inhalatoria has a positive influence on microbiological air quality. Owing to a unique microclimate and low air contamination, Ciechocinek and Inowroclaw comply with all necessary requirements set for health resorts specializing in treating upper respiratory tract infections.

Extracellular enzyme activity in a willow sewage treatment system.

This paper presents the results of studies on the activity of extra-cellular enzymes in soil-willow vegetation filter soil which is used in the post-treatment of household sewage in an onsite wastewater treatment system located in central Poland. Wastewater is discharged from the detached house by gravity into the onsite wastewater treatment system. It flows through a connecting pipe into a single-chamber septic tank and is directed by the connecting pipe to a control well to be further channelled in the soil-willow filter by means of a subsurface leaching system. Soil samples for the studies were collected from two depths of 5 cm and 1 m from three plots: close to the wastewater inflow, at mid-length of the plot and close to its terminal part. Soil samples were collected from May to October 2009. The activity of the extra-cellular enzymes was assayed by the fluorometric method using 4-methylumbelliferyl and 7-amido-4-methylcoumarin substrate. The ranking of potential activity of the assayed enzymes was the same at 5 cm and 1 m soil depths, i.e. esterase > phosphmomoesterase > leucine-aminopeptidase > ß-glucosidase > α-glucosidase. The highest values of enzymatic activity were recorded in the surface layer of the soil at the wastewater inflow and decreased with increasing distance from that point.

Production of antifungal chitinase by Aspergillus niger LOCK 62 and its potential role in the biological control.

Aspergillus niger LOCK 62 produces an antifungal chitinase. Different sources of chitin in the medium were used to test the production of the chitinase. Chitinase production was most effective when colloidal chitin and shrimp shell were used as substrates. The optimum incubation period for chitinase production by Aspergillus niger LOCK 62 was 6 days. The chitinase was purified from the culture medium by fractionation with ammonium sulfate and affinity chromatography. The molecular mass of the purified enzyme was 43 kDa. The highest activity was obtained at 40 °C for both crude and purified enzymes. The crude chitinase activity was stable during 180 min incubation at 40 °C, but purified chitinase lost about 25 % of its activity under these conditions. Optimal pH for chitinase activity was pH 6-6.5. The activity of crude and purified enzyme was stabilized by Mg(2+) and Ca(2+) ions, but inhibited by Hg(2+) and Pb(2+) ions. Chitinase isolated from Aspergillus niger LOCK 62 inhibited the growth of the fungal phytopathogens: Fusarium culmorum, Fusarium solani and Rhizoctonia solani. The growth of Botrytis cinerea, Alternaria alternata, and Fusarium oxysporum was not affected.

Identification and characterization of a chitinase of Stenotrophomonas maltophilia, a bacterium that is antagonistic towards fungal phytopathogens.

A rhizosphere strain of Stenotrophomonas maltophilia strain MUJ that is strongly antagonistic towards fungal phytopathogens secretes to the culture medium a single form of active chitinolytic enzyme belonging to family 18 of glycosyl hydrolases. The chitinase was purified by a two-stage procedure embracing fractionation with ammonium sulfate and affinity chromatography. The molecular mass of the purified enzyme determined by SDS-PAGE was approximately 52 kDa. The enzyme demonstrated highest activity at 45°C and pH 6.8. The enzymatic protein showed considerable thermal stability during 2 h incubation at 45°C. The activity of the enzyme was strongly inhibited in the presence of Hg²âº and Cu²âº. By applying mass spectrometry analysis, the peptides derived from the purified chitinase were assigned to amino acid sequences of the type ChiA chitinases synthesized by Stenotrophomonas bacteria. The purified enzyme inhibited the growth of fungal phytopathogens belonging to the genera Fusarium, Rhizoctonia and Alternaria.

Influence of the root system of the Common Osier (Salix viminalis L.) on abundance of heterotrophic bacteria in the willow sewage treatment system.

The impact of the Common Osier (Salix viminalis L.) root system on number (CFU) of heterotrophic bacteria and their production in a soil-willow filter was examined. The Osier rhizosphere was found to be suitable habitat for growth of the examined microbial group, and the root system stimulated development of heterotrophic bacteria. The rhizosphere bacteria to control soil bacteria (R:C) ratio oscillated between 2.48 and 2.75 depending on the location of sample collection. The highest abundance of bacteria as well as highest bacterial production was observed at location I, near sewage discharge onto the plot. There was a significant positive correlation between the number of heterotrophic bacteria and the bacterial production.

Occurrence and activity of microorganisms in shrimp waste.

This study aimed to determine the occurrence and respiration activity of heterotrophic bacteria and fungi in shrimp shell waste and to evaluate the role of chitinolytic bacteria and fungi in its decomposition. The highest levels of bacteria were found in shrimp heads sections and the lowest in exoskeletons. The level of fungi was much lower, with the highest proportion present in heads sections and the lowest in exoskeletons. Chitinolytic bacteria constituted a small percentage of the total heterotrophic bacteria in fresh shrimp waste, averaging 4% in exoskeletons, 2.4% in all parts, and 2% in heads. No chitinolytic bacteria were detected in stored waste. In contrast, the percentage of chitinolytic fungi in shrimp waste was much higher than that of bacteria. Chitinolytic fungi constituted 25-60% of the total fungi in fresh waste and 15-40% in stored waste. Chitinolytic bacteria isolated from heads sections were characterized by the highest chitinolytic activity, averaging 11.2 nmol of methylumbelliferyl x mg(-1) protein x h(-1), whereas the lowest activity was in strains from exoskeletons, averaging 3.2 nmol of methylumbelliferyl x mg(-1) protein x h(-1). The chitinolytic activity of fungi isolated from all parts waste, head sections, and exoskeletons was similar. The respiration activity of microorganisms in fresh and stored waste was similar. Oxygen consumption activity increased during incubation and approached a saturation value between days 4 and 5. No correlation between the end value of respiratory activity in the analyzed section of shrimp discard after 5 days and the level of bacteria and fungi was observed. The only significant correlation observed was between the respiratory activity of the shrimp and the level of fungi. The respiration activity significantly depended on the analyzed section of shrimp discard (p<0.000).