Isolation and identification of rhizospheric pseudomonads with insecticidal effects from various crops in Khuzestan Province, Iran.

Pseudomonas bacteria include a variety of species with distinct characteristics. Some species within this genus are known for their ability to stimulate plant growth. Recently, the potential of these bacteria in controlling insect pests has been documented. In this study, 58 bacterial isolates were purified from rhizospheres of wheat, broad bean and canola that were collected from different fields of Khuzestan province in south-west of Iran. With biochemical tests 19 non plant pathogenic pseudomonads strains were detected and their lethal effects on the eggs and larvae of Ephestia keuhniella as an important pest that infests stored products, were evaluated under laboratory conditions. For the bioassays, two concentrations of each strain were administered, and the 5th instar larvae and eggs of the pest were subjected to treatment. Mortality rates were recorded after 24 h. The results showed that all isolated Pseudomonad strains of this study had insecticidal effects against eggs and larvae of E. keuhniella. The strains AWI1, AWI2, AWI7, ABI12, ABI15 and ABI16 displayed the highest mortality rate (91.1 %, 86.2 %, 82.3 %, 84.2, 90.5 % and 90.5 %, respectively). Molecular identification and phylogeny tree according to 16 s rRNA sequencing clarified that AWI1, AWI2 belong to P. plecoglossicida, AWI5 belongs to P. lini, ABI12, ABI15 and ABI16 belong to P. taiwanensis. Moreover, the bacterial efficacy at a suspension concentration of 0.5 OD (80 %) was significantly greater than that at a concentration of 0.2 OD (63.33 %). No significant difference was detected in the response of the pest larvae or eggs to the different strains. Furthermore, olfactory trials revealed that the female parasitoid wasp Habrabracon hebetor actively avoided the infection of the treated larvae by the strains. These findings have practical implications for the development of microbiological pest control strategies.

Community-based mechanisms underlying the root cadmium uptake regulated by Cd-tolerant strains in rice (Oryza sativa. L).

In recent years, the problem of Cd pollution in paddy fields has become more and more serious, which seriously threatens the safe production of food crops and human health. Using microorganisms to reduce cadmium pollution in rice fields is a green, safe and efficient method, the complicated interactions between the microbes in rice roots throughout the process of cadmium absorption by rice roots are poorly understood. In this investigation, a hydroponic pot experiment was used to examine the effects of bacteria R3 (Herbaspirillum sp) and T4 (Bacillus cereus) on cadmium uptake and the endophytic bacterial community in rice roots. The results showed that compared with CK (Uninoculated bacterial liquid), the two strains had significant inhibitory or promotive effects on cadmium uptake in rice plant, respectively. Among them, the decrease of cadmium content in rice plants by R3 strain reached 78.57-79.39%, and the increase of cadmium content in rice plants by T4 strain reached 140.49-158.19%. Further investigation showed that the cadmium content and root cadmium enrichment coefficient of rice plants were significantly negatively correlated with the relative abundances of Burkholderia and Acidovorax, and significantly positively correlated with the relative abundances of Achromobacter, Agromyces and Acidocella. Moreover, a more complex network of microbes in rice roots inhibited rice plants from absorbing cadmium. These results suggest that cadmium uptake by rice plants is closely related to the endophytic bacterial community of roots. This study provides a reference scheme for the safe production of crops in cadmium contaminated paddies and lays a solid theoretical foundation for subsequent field applications.

A novel use of Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae) as inoculative agent of baculoviruses.

BACKGROUND: Alphabaculoviruses are Lepidoptera-specific virulent pathogens that infect numerous pests, including the Spodoptera complex. Due to their low environmental persistence, the traditional use of Alphabaculoviruses as bioinsecticides consist in high-rate spray applications with repeated treatments. Several abiotic and biotic factors can foster its dispersion, promoting their persistence in the agroecosystem. Amongst biotic factors, predatory arthropods can disperse the viruses by excretion after preying on infected individuals. Therefore, this study focused on promoting predator's ingestion of nucleopolyhedrovirus (NPV)-treated diets, and the later exposition of the insect host to leaf surfaces contaminated with predator excreta. The virus-host-predator system studied was Spodoptera littoralis nucleopolyhedrovirus (SpliNPV), Spodoptera littoralis (Boisduval) and Nesidiocoris tenuis (Reuter). The infective potential of N. tenuis feces and the retention time of SpliNPV were assessed under laboratory conditions after feeding on treated diets (sucrose solution and Ephestia kuehniella eggs). RESULTS: Mortality of S. littoralis larvae was lower via N. tenuis excretion than in positive control (spray application) in the first infection cycle, together with a delay in host death. In the second infection cycle, both SpliNPV-treated diets triggered 100% mortality. Both diets allowed the transmission of SpliNPV, with a faster excretion via sucrose solution compared to E. kuehniella eggs. SpliNPV remained in N. tenuis digestive tract and was viable after excretion at least for 9 days for both diets. CONCLUSIONS: This study demonstrated the potential of the predator N. tenuis as inoculative agent of baculoviruses, representing a new alternative that, along with inundative applications, might contribute to improve pest management strategies. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

MALDI-TOF as a tool for microbiological monitoring in areas considered aseptic.

To maintain asepsis in production environments, contamination must be constantly controlled. To this end, microbiological monitoring is constantly used with the objective of evaluating the incidence of microorganisms prevalent in the sampling of air, surface, and people, in the area of an environment considered aseptic, isolated, and identified using the rapid and automated phenotypic microbiological methodology, highlighting the MALDI-TOF mass spectrometry analysis technique (MS), being identified at the level of genus and/or species. For that purpose, microbiological control of environmental monitoring of environments considered aseptic in a pharmaceutical industry was conducted for 12 months. The isolated microorganisms were identified using the mass spectrometry identification method (MALDI-TOF). In area classification A, the most prevalent microorganisms were bacteria in the sampling person. The microbial population was composed of bacteria of the genus Micrococcus sp. and Staphylococcus sp. Based on the results, it is possible to observe that in an environment where the process requires human operations, possible microbial contamination is inevitable and requires the identification of microorganisms at least at the level of species and/or genus. The microorganisms identified and found in the sampling of the aseptic environment must be evaluated with frequency to ensure that the productive environment guarantees the quality of the product produced.

Microbiological Aspects of Pharmaceutical Manufacturing of Adipose-Derived Stem Cell-Based Medicinal Products.

Subcutaneous adipose tissue is an excellent source of mesenchymal stem cells (ADSCs), which can be used in cell therapies as an active substance in advanced therapy medicinal products (ATMPs). Because of the short shelf-life of ATMPs and the time needed to obtain the results of microbiological analysis, the final product is often administered to the patient before sterility is confirmed. Because the tissue used for cell isolation is not sterilized to maintain cell viability, controlling and ensuring microbiological purity at all stages of production is crucial. This study presents the results of monitoring the contamination incidence during ADSC-based ATMP manufacturing over two years. It was found that more than 40% of lipoaspirates were contaminated with thirteen different microorganisms, which were identified as being physiological flora from human skin. Such contamination was successfully eliminated from the final ATMPs through the implementation of additional microbiological monitoring and decontamination steps at various stages of production. Environmental monitoring revealed incidental bacterial or fungal growth, which did not result in any product contamination and was reduced thanks to an effective quality assurance system. To conclude, the tissue used for ADSC-based ATMP manufacturing should be considered contaminated; therefore, good manufacturing practices specific to this type of product must be elaborated and implemented by the manufacturer and the clinic in order to obtain a sterile product.

Genetic variability of Metarhizium isolates from the Ticino Valley Natural Park (Northern Italy) as a possible microbiological resource for the management of Popillia japonica.

The natural occurrence of entomopathogenic fungi (EPF) was investigated along the Ticino River (Ticino River Natural Park, Novara Province, Piedmont, Italy), at the center of the area of the first settlement of the invasive alien pest Popillia japonica. Using Zimmermann's "Galleria bait method", EPF were successfully isolated from 83 out of 155 soil samples from different habitats (perennial, cultivated, or uncultivated meadows, woodlands, and riverbanks). Sequencing of the 5' end of the Translation Elongation Factor 1 alfa (5'-TEF) region allowed the assignment of 94% of the isolates to Metarhizium spp., while 8% and 7% were assigned to Beauveria spp. and Paecilomyces spp., respectively. Four Metarhizium species were identified: Metarhizium robertsii was the most common one (61.5% of the isolates), followed by M. brunneum (24.4%), M. lepidiotae (9%), and M. guizhouense (5.1%). Microsatellite marker analysis of the Metarhizium isolates revealed the presence of 27 different genotypes, i.e., 10 genotypes among M. robertsii, 8 among M. brunneum, 5 among M. lepidiotae, and 4 among M. guizhouense. Metarhizium brunneum appeared to be associated with woodlands and more acid soils, while the other species showed no clear association with a particular habitat. Laboratory virulence tests against P. japonica 3rd instar larvae allowed the identification of one M. robertsii isolate that showed efficacy as high as 80.3%. The importance of this kind of study in the frame of eco-friendly microbiological control is discussed.

Physico-Chemical and Microbiological Control of the Composting Process of the Organic Fraction of Municipal Solid Waste: A Pilot-Scale Experience.

The aim of this work was to carry out a pilot experiment to monitor OFMSW (organic fraction of municipal solid waste) composting processes using different types of installations (automatic reactor, aerated static pile and turned pile). To carry out the process, pruning waste was used as structuring material (SM), in a 1:1 and 1:2, v:v, OFMSW:SM ratio. Monitoring was carried out through the control of physico-chemical and microbiological parameters, such as temperature, pH, humidity, Rottegrade, Solvita tests, the presence of Salmonella sp. and Escherichia coli, total coliform, and Enterococcus sp. concentrations. After carrying out the tests, it can be affirmed that the three types of installations used worked correctly in terms of the monitoring of physico-chemical parameters, giving rise to a compost of sufficient stability and maturity to be applied on agricultural soil. In all cases the bacterial concentrations in the final compost were lower than those detected in the mixture of initial components for its preparation, thus complying with the requirements established in RD 506/2013 and RD 999/2017RD on fertilizer products. However, it cannot be affirmed that one of the three types of installation used produces a greater bacterial inactivation than the others. When composting with different types of facilities, it is of interest to optimize the irrigation and aeration system in order to have a better control of the process and to study the possible temperature gradients in the piles to ensure good sanitization without the risk of bacterial proliferation a posteriori. Finally, the different initial mixtures of OFMSW and SM used in this study did not have a significant influence on the functioning of the composting process or on the microbiological quality during the process. The irrigation water can provide a bacterial contribution that can lead to increases in concentration during the composting process. This study is part of the Life-NADAPTA project (LIFE16 IPC/ES/000001), an integrated strategy for adaptation to Climate Change in Navarra, where NILSA participates in water action and collaborates in agricultural action, which includes among its objectives the development of new soil amendments from different organic waste.

Dermatocosmetic Emulsions Based on Resveratrol, Ferulic Acid and Saffron (Crocus sativus) Extract to Combat Skin Oxidative Stress-Trigger Factor of Some Potential Malignant Effects: Stability Studies and Rheological Properties.

The increasing incidence of skin diseases, against the background of increased pollution, urbanism, poor habits in lifestyle, work, rest, diet and general medication, led to the development of products with a protective effect. These new types of dermatocosmetic preparations ensure maximum benefits with minimal formulation. Antioxidants are, nowadays, ingredients that stand out with a proven role in skin protection from oxidative stress and its effects. Thus, research has shown that light-textured formulas, quickly absorbed into the skin, with optimum hydration and protection against excessive free radicals, uphold the skin integrity and appearance. This article aims to evaluate essential criteria for a newly marketed product: stability, rheological properties and microbiological characteristics of oil-in-water emulsions based on a mixture of 3% resveratrol 0.5% ferulic acid and 1mL alcoholic extract of Saffron. The tests led to the conclusion that O / W dermatocosmetic emulsions, based on 3% resveratrol and 0.5% ferulic acid, or also 1mL alcoholic extract of Saffron, show resistance to microbiological contamination, good rheological properties (viscoelastic behavior, structural stability, acceptable shearing behavior) that reveal satisfactory texture and high physical stability during storage. These results encourage the transition to dermatological testing as the final stage in considering a new commercial product.

Microbiological Safety and Quality of Meals and Work Surfaces in Collective Catering Systems in Central Italy: A Five-Year Monitoring Study.

Ready-to-eat (RTE) meals produced and served by the catering system still represent one of the major causes of foodborne outbreaks, especially for susceptible consumers. Despite the great progress in food hygiene and safety, the systematic monitoring of microbial contamination of foodstuff is the most effective tool to ensure food safety and protect consumers' health. The aim of this study was to perform a thorough assessment of the microbial safety and quality of meals and work surfaces of collective catering systems in central Italy, over a five-year period (2014−2018). In total 11,012 microbiological analytical determinations were performed in food matrices (80.1%) and environmental samples (19.9%). The results obtained show a low level of non-conformities ranging from 2.2% to 6.3% of total samples, concerning both hygiene and safety parameters. A decreasing trend of non-conformities during the years was also highlighted (p-value < 0.05), especially for environmental samples. This study suggests that the implementation of Good Manufacturing Practices (GMPs) and the proper definition of Hazard Analysis and Critical Control Point (HACCP) plans, combined with a thorough evaluation of microbiological monitoring, are able to ensure high levels of food safety and hygiene.

Quality of cosmetics with active caffeine in cream and gel galenic bases prepared by compounding pharmacies / Qualidade de cosméticos com cafeína ativa em bases galênicas em creme e gel preparadas por farmácias de manipulação

As there is a great scarcity of studies on the importance of good compounding practices in the preparation of cosmetics, this study aimed to evaluate the quality control of cosmetics with active ingredient caffeine for the treatment of cellulite prepared by magistrals pharmacies. Microbiological analyzes, pH determination, color measurement, quantification of the percentage of the active ingredient caffeine and viscosity in creams and gels with 5% of the active ingredient caffeine were performed. In the microbiological analysis, the presence of molds and yeasts was verified above the permitted level according to the Brazilian pharmacopoeia. The pH decreased over time, contributing to the formulations becoming more acidic. In the color parameters, it was found that pharmacy F4 showed a brownish color, both for the gel and for the cream. The percentage of caffeine was within specifications in all formulations and the viscosity remained unchanged during the shelf life of the samples. It is important that the compounding pharmacies demand more effectively the commitment of the team, as well as the analysis of the raw material according to the microbiological control regulations to translate into the quality of the products prepared by the pharmacies and favor the consumer in the effective objective that the product you want to achieve.

Como há uma grande escassez de estudos sobre a importância de boas práticas de manipulação na elaboração de cosméticos, o presente estudo teve como objetivo avaliar o controle de qualidade de cosméticos com princípio ativo cafeína para o tratamento de celulite, elaborados por farmácias de manipulação de Campo Mourão. Foram realizadas análises microbiológicas, determinação do pH, mensuração da coloração, quantificação da porcentagem do princípio ativo cafeína e viscosidade em cremes e geís com 5% do princípio ativo cafeína. Nas análises microbiológicas foi verificada a presença de bolores e leveduras acima do pemitido segundo a farmacopéia brasileira. O pH diminuiu ao longo do tempo, contribuindo para que as formulações ficassem mais ácidas. Nos parâmetros de cor averiguou-se que a farmácia F4 apresentou uma coloração amarronzada, tanto para o gel quanto para o creme. A porcentagem de cafeína estava dentro das especificações em todas as formulações e a viscosidade se manteve inalterada durante o prazo de validade das amostras. Com o propósito de um melhor preparo magistral é importante que as farmácias de manipulação exijam de forma mais eficaz o comprometimento da equipe, bem como a análise da matéria-prima segundo as regulamentações de controle microbiológico. Além disso, a implementação de normas mais rígidas e o melhor controle da matéria-prima e das formulações finais fazem com que a qualidade dos produtos manipulados pelas farmácias magistrais seja aumentada significativamente, favorecendo o consumidor na efetiva finalidade a que o produto objetiva atingir.

Quality of cosmetics with active caffeine in cream and gel galenic bases prepared by compounding pharmacies

Abstract As there is a great scarcity of studies on the importance of good compounding practices in the preparation of cosmetics, this study aimed to evaluate the quality control of cosmetics with active ingredient caffeine for the treatment of cellulite prepared by magistrals pharmacies. Microbiological analyzes, pH determination, color measurement, quantification of the percentage of the active ingredient caffeine and viscosity in creams and gels with 5% of the active ingredient caffeine were performed. In the microbiological analysis, the presence of molds and yeasts was verified above the permitted level according to the Brazilian pharmacopoeia. The pH decreased over time, contributing to the formulations becoming more acidic. In the color parameters, it was found that pharmacy F4 showed a brownish color, both for the gel and for the cream. The percentage of caffeine was within specifications in all formulations and the viscosity remained unchanged during the shelf life of the samples. It is important that the compounding pharmacies demand more effectively the commitment of the team, as well as the analysis of the raw material according to the microbiological control regulations to translate into the quality of the products prepared by the pharmacies and favor the consumer in the effective objective that the product you want to achieve.

Resumo Como há uma grande escassez de estudos sobre a importância de boas práticas de manipulação na elaboração de cosméticos, o presente estudo teve como objetivo avaliar o controle de qualidade de cosméticos com princípio ativo cafeína para o tratamento de celulite, elaborados por farmácias de manipulação de Campo Mourão. Foram realizadas análises microbiológicas, determinação do pH, mensuração da coloração, quantificação da porcentagem do princípio ativo cafeína e viscosidade em cremes e geís com 5% do princípio ativo cafeína. Nas análises microbiológicas foi verificada a presença de bolores e leveduras acima do pemitido segundo a farmacopéia brasileira. O pH diminuiu ao longo do tempo, contribuindo para que as formulações ficassem mais ácidas. Nos parâmetros de cor averiguou-se que a farmácia F4 apresentou uma coloração amarronzada, tanto para o gel quanto para o creme. A porcentagem de cafeína estava dentro das especificações em todas as formulações e a viscosidade se manteve inalterada durante o prazo de validade das amostras. Com o propósito de um melhor preparo magistral é importante que as farmácias de manipulação exijam de forma mais eficaz o comprometimento da equipe, bem como a análise da matéria-prima segundo as regulamentações de controle microbiológico. Além disso, a implementação de normas mais rígidas e o melhor controle da matéria-prima e das formulações finais fazem com que a qualidade dos produtos manipulados pelas farmácias magistrais seja aumentada significativamente, favorecendo o consumidor na efetiva finalidade a que o produto objetiva atingir.

Quality of cosmetics with active caffeine in cream and gel galenic bases prepared by compounding pharmacies / Qualidade de cosméticos com cafeína ativa em bases galênicas em creme e gel preparadas por farmácias de manipulação

As there is a great scarcity of studies on the importance of good compounding practices in the preparation of cosmetics, this study aimed to evaluate the quality control of cosmetics with active ingredient caffeine for the treatment of cellulite prepared by magistrals pharmacies. Microbiological analyzes, pH determination, color measurement, quantification of the percentage of the active ingredient caffeine and viscosity in creams and gels with 5% of the active ingredient caffeine were performed. In the microbiological analysis, the presence of molds and yeasts was verified above the permitted level according to the Brazilian pharmacopoeia. The pH decreased over time, contributing to the formulations becoming more acidic. In the color parameters, it was found that pharmacy F4 showed a brownish color, both for the gel and for the cream. The percentage of caffeine was within specifications in all formulations and the viscosity remained unchanged during the shelf life of the samples. It is important that the compounding pharmacies demand more effectively the commitment of the team, as well as the analysis of the raw material according to the microbiological control regulations to translate into the quality of the products prepared by the pharmacies and favor the consumer in the effective objective that the product you want to achieve.

Como há uma grande escassez de estudos sobre a importância de boas práticas de manipulação na elaboração de cosméticos, o presente estudo teve como objetivo avaliar o controle de qualidade de cosméticos com princípio ativo cafeína para o tratamento de celulite, elaborados por farmácias de manipulação de Campo Mourão. Foram realizadas análises microbiológicas, determinação do pH, mensuração da coloração, quantificação da porcentagem do princípio ativo cafeína e viscosidade em cremes e geís com 5% do princípio ativo cafeína. Nas análises microbiológicas foi verificada a presença de bolores e leveduras acima do pemitido segundo a farmacopéia brasileira. O pH diminuiu ao longo do tempo, contribuindo para que as formulações ficassem mais ácidas. Nos parâmetros de cor averiguou-se que a farmácia F4 apresentou uma coloração amarronzada, tanto para o gel quanto para o creme. A porcentagem de cafeína estava dentro das especificações em todas as formulações e a viscosidade se manteve inalterada durante o prazo de validade das amostras. Com o propósito de um melhor preparo magistral é importante que as farmácias de manipulação exijam de forma mais eficaz o comprometimento da equipe, bem como a análise da matéria-prima segundo as regulamentações de controle microbiológico. Além disso, a implementação de normas mais rígidas e o melhor controle da matéria-prima e das formulações finais fazem com que a qualidade dos produtos manipulados pelas farmácias magistrais seja aumentada significativamente, favorecendo o consumidor na efetiva finalidade a que o produto objetiva atingir.

Morphological and molecular identification of four isolates of the entomopathogenic fungal genus Akanthomyces and their effects against Bemisia tabaci on cucumber.

The cotton whitefly, Bemisia tabaci Gen. (Hem., Aleyrodidae), is a key pest of many vegetables. Entomopathogenic fungi are promising microbial control agents against B. tabaci, but limited information is available concerning indigenous Iranian isolates. In this study, three isolates of Akanthomyces lecanii (PAL6, PAL7, and PAL8) and one isolate of A. muscarius (AGM5) were obtained from citrus hemipteran pests, Pulvinaria aurantii Cock. and Aphis gossypii Glover, in Mazandaran province, northern Iran. The isolates were then morphologically and molecularly identified. The efficacies of five different agar media for vegetative growth and conidiation of each isolate were determined. Potato dextrose agar was the medium on which the fungal mycelia developed at a relatively high rate. However, the highest rate of conidiation was found on Sabouraud dextrose agar. To determine the effects of the isolates on B. tabaci, a dose-response bioassay was carried out to estimate lethal concentration (LC50) and lethal time (LT50) values of each fungal isolate to second instar nymphs. The mean LC50 values of A. lecanii isolates ranged from 4.22 × 106 to 7.35 × 1013 conidia ml-1 at 5 to 7 days after the treatment. For A. muscarius, the values varied from 9.2 × 104 to 8.7 × 1010 conidia ml-1 at 5 to 7 days after the treatment. The lowest and the highest mean LC50 values were observed for A. mucarius (AGM5) and A. lecanii (isolate PAL6), respectively. The mean LT50 values of A. lecanii and A. muscarius isolates were 7.1-9.0 and 4.9-7.2 days, respectively. The LT50 values of A. muscarius were significantly lower than the other isolates. Overall, all isolates, especially A. muscarius (AGM5), exhibited appropriate potential as a biological control agent against B. tabaci.

Combination of ceftaroline and daptomycin as treatment for complicated osteomyelitis.

Osteomyelitis is an infection involving bone. Staphylococcus aureus is the pathogen most frequently implicated; less frequently involved are other gram-positive organisms, such as Staphylococcus epidermidis, and also gram-negative organisms. The antibiotic of choice for treatment of osteomyelitis caused by methicillin-resistant staphylococci (MRS) is vancomycin, although other alternatives such as daptomycin or teicoplanin are also considered. Osteomyelitis caused by MRS can be difficult to treat safely and effectively. This case report describes the successful use of daptomycin combined with ceftaroline for the treatment of osteomyelitis caused by methicillin-resistant S. epidermidis (MRSE) in a 54-year-old woman, emphasising the clinical pharmacist's role in antimicrobial stewardship programmes. This alternative combination has been studied in the treatment of methicillin-resistant S. aureus (MRSA), but it may also be useful in MRSE.

Quality control and immunomodulatory potential for clinical-grade equine bone marrow-derived mesenchymal stromal cells and conditioned medium.

This study aimed to assess the safety and reproducibility of cell therapy for its use in clinical practice. We performed immunophenotypic characterization of equine bone marrow-derived mesenchymal stromal cells (BMMSCs) by flow cytometry using CD90, CD19, CD14, CD105, CD45, and HLA-DR markers (n = 4); GTG banding cytogenetic analysis (n = 3); and microbiological quality control (n = 4). The immunomodulatory potentials of BMMSCs (n = 4) and its conditioned medium (CM, n = 3) were investigated by in vitro lymphocyte inhibition assay using phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMCs). BMMSCs populations isolated from all animals showed high expression of CD90 and CD105, and low expression of CD19, CD4, CD45, and HLA-DR. Of the 60 metaphases analyzed, 5% presented aneuploidy on random chromosomes and no contamination was found based on microbiological analyses. Both treatments significantly inhibited lymphocyte proliferation (> 50%), compared with PHA-stimulated PBMCs (p < 0.0001). These promising results for BMMSCs and CM justify their potential as a therapeutic approach for inflammatory diseases. The techniques used in this study were effective in assessing the quality and determining the minimum criteria for the clinical use of BMMSCs in veterinary medicine.

Microbiological Control of Cellular Products: The Relevance of the Cellular Matrix, Incubation Temperature, and Atmosphere for the Detection Performance of Automated Culture Systems.

BACKGROUND: The microbiological control of cellular products sometimes causes significant procedural issues for quality control laboratories. According to the European Pharmacopoeia (EP), the microbiological control of cellular products requires a 7- to 14-day incubation period at two different incubation temperatures using aerobic and anaerobic growth media. However, the suitability of these test conditions for efficient quality control can be influenced by many conditions, such as the expected microbial spectrum of contamination or the texture and composition of the cellular product. Because of interference, direct inoculation and membrane filtration as reference methods of pharmacopoeia are largely unsuitable for the microbiological control of cellular products; therefore, alternative and, above all, automated methods are the focus of interest. OBJECTIVE: The aim of our study was to evaluate the method suitability and possible effects of cell matrix, incubation temperature, and oxygen pressure on the detection performance of automated culture systems. METHODS: The BacT/ALERT® 3DTM Dual T system (bioMérieux, Nürtingen, Germany) was used to evaluate the factors influencing automated microbiological control of cellular products. The tests were performed using microbial strains recommended by the EP for microbiological method suitability testing and additional relevant possible contaminants of human-derived stem-cell products under varying culture and cell matrix conditions. RESULTS: All contaminants were detected by the system in the required period of 2-5 days. Low incubation temperatures (22°C) had overall negative effects on the detection kinetics of each type of microbial contamination. The adverse effects of the accompanying cell matrix on the detection properties of the system could be compensated in our study by incubation at 32°C in both the aerobic and the anaerobic culture conditions. CONCLUSION: Automated culture techniques represent a sufficient approach for the microbiological control of cellular products. The negative effects of the cell matrix and microbial contamination on the detection performance can be compensated by the application of variable culture conditions in the automated culture system.

Preventative Approach to Microbial Control of Capnodis tenebrionis by Soil Application of Metarhizium brunneum and Beauveria bassiana.

Management of the Mediterranean flat-headed root-borer, Capnodis tenebrionis, is critical due to the larvae's root localization. Neonate larvae can be exposed to natural enemies before penetrating the roots. Application of Metarhizium brunneum strain Mb7 and Beauveria bassiana strain GHA formulations on rice granules was investigated for their efficacy against C. tenebrionis larvae. Mb7 application, evaluated on apricot twigs, significantly and dose-dependently reduced colonization rates of neonates, with highest mortality at 108 conidia/g soil. Neonate susceptibility to Mb7 and GHA was evaluated on potted rootstocks (GF677 almond × peach, 2729 plum) planted in entomopathogenic fungi (EPF)-premixed soil (1.3-1.6 × 105 conidia/cm3 soil) or in EPF-free soil surface-treated with 5 g Mb7 fungal granules (1.25 × 109 conidia). Larval colonization rates were reduced 7.4-fold in 2729 by both fungi; only Mb7 completely prevented colonization of GF677 by larvae. Larvae inside plant galleries exhibited mycosis with EPF-treated soils and both fungi proliferated on larval frass. Mb7 conidia germinated in the rhizosphere of GF677, and conidia of both fungi remained viable throughout the trial. Galleria baiting technique was used on EPF-treated soil to evaluate EPF infectivity over time; Mb7 and GHA persisted 180 and 90 days post inoculation, respectively. The formulation (fungus-covered rice grains), delivery method (mixing with soil) and persistence (3-6 months) of Mb7 and GHA are feasible for potential field application to control C. tenebrionis.

The Effect of High-Intensity Ultrasound on the Physicochemical and Microbiological Properties of Mexican Panela Cheese.

High-intensity ultrasound could be an alternative to pasteurization for cheeses made with fresh raw milk, the properties of which must be preserved as part of their intangible cultural heritage, such as Panela cheese in Mexico. This research aimed to study the effect of the amplitude (50% and 100%) and application time (0, 5, and 10 min) of ultrasound treatment of fresh raw milk, on the yield and microbiological and physicochemical qualities of Panela cheese after 24 h of storage at 4 °C. The yield was increased to 24.29% with 10 min of ultrasonication, although the amount of exudate was higher in the ultrasonic product than in the control (20.33%). As the ultrasonication time increased, the yellowness (b*) increased significantly, while the hue angle decreased (with values close to 90°), resulting in evident yellow tones in cheeses made with milk treated for 10 min. The pH significantly increased from 6.6 to 6.74 with 5 min of ultrasound, but decreased to 6.37 with 10 min of ultrasonication. Although no significant differences were found in fat content, the protein significantly increased with 5 min of sonication, but it decreased markedly when ultrasound was applied for 10 min. Ultrasound treatment with amplitudes of 50% effectively decreased the counts of coliform bacteria regardless of ultrasonication time. However, the mesophilic bacteria increased by a 0.9 log with an amplitude of 100% and 10 min treatment. The results showed that ultrasound improved the yield and microbial, nutritional, and physicochemical properties of Panela cheese.

Influence of Bacillus thuringiensis application timing on population dynamics of gypsy moth in Mediterranean cork oak forests.

BACKGROUND: The gypsy moth, Lymantria dispar, is one of the main pests of oak forests worldwide and causes extensive defoliation during its periodic outbreaks. In the Mediterranean region, control of gypsy moth populations in cork oak forests is based on application of Bacillus thuringiensis serovar kurstaki (Btk) formulations. This research investigated the effects of Btk applications carried out in two different population development phases on gypsy moth population dynamics. With this aim, temporal and spatial fluctuation patterns of L. dispar egg density were monitored in cork oak forests treated with Btk applications from 2004 to 2009 in Sardinia (Italy). RESULTS: Applications undertaken during the progradation and culmination phases protected the oak canopies equally in the year of application, leading to a similar decrease in pest population density in the following year. However, the medium-term effectiveness of Btk differed between the two application timings, because only applications in the culmination phase caused a gradual decrease in L. dispar infestations throughout subsequent years. By contrast, when the application was undertaken during the progradation phase, population density increased again after 2-3 years. Moreover, Btk applications in the culmination phase reduced significantly the number of years in which gypsy moth density was damaging compared with those done in progradation. CONCLUSIONS: Our results indicate that Btk applications during the culmination phase were more effective than those in the progradation period, because application in the latter case did not suppress the population, but only postponed the outbreak peak by 2-3 years. © 2019 Society of Chemical Industry.

[Microbiological testing of flexible endoscopes: contribution of the approach of accreditation in the implementation of regulation]. / Analyse microbiologique des solutions de contrôle d'endoscope : apport de la démarche d'accréditation dans la mise en application de la réglementation.

The microbiological surveillance of flexible endoscopes reprocessing involves counts of the mesophilic total aerobic revivable flora and the detection of indicator microorganisms of dysfunction. We presented the assay at the COFRAC certification, to ensure a maximal level of confidence in the quality of our results. The existing quality system management was used by the type B widened flexible scope for data from previous accreditations for the identification of Staphylococcus aureus, Enterococci and Enterobacteriaceae. The quality insurance of the results and the technical authorization were guaranteed by the participation at a program of interlaboratory comparisons and the elaboration of internal quality controls. Risks analyses and conventions were wrote with endoscopy sectors. The visit led the opening of a single and not critical deviation sheet on the control of consumables and the accreditation for the analysis was pronounced.