Chicken IgY Antibodies Provide Mucosal Barrier against SARS-CoV-2 Virus and Other Pathogens.

BACKGROUND: This mini review includes two case descriptions. It introduces the use of chicken egg yolk antibody (IgY) solutions in the prevention and cure of viral and bacterial infections. Application for the protection against severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), rotavirus, and influenza viruses, as well as for the eradication of Pseudomonas aeruginosa, caries, various enteric bacteria and other pathogens, and toxins have been developed. This approach is a fast, reliable, safe, and tested method for producing molecular shield and protection against emerging pathogens and epidemics. In the current pandemic situation caused by coronavirus disease-2019 (COVID-19), this method of passive immunization could be applied for rapid protection against modifiable agents. The specific IgY antibodies start to accumulate into egg yolks about 3 weeks after the immunization of the chicken. The product can be collected safely, as the antigen is not found in the eggs. This method for microbial safety uses natural means and commonly used food substances, which have been tested and could be produced for both blocking epidemics and applying personalized medicine.

The second International Symposium on Fungal Stress: ISFUS.

The topic of 'fungal stress' is central to many important disciplines, including medical mycology, chronobiology, plant and insect pathology, industrial microbiology, material sciences, and astrobiology. The International Symposium on Fungal Stress (ISFUS) brought together researchers, who study fungal stress in a variety of fields. The second ISFUS was held in May 8-11 2017 in Goiania, Goiás, Brazil and hosted by the Instituto de Patologia Tropical e Saúde Pública at the Universidade Federal de Goiás. It was supported by grants from CAPES and FAPEG. Twenty-seven speakers from 15 countries presented their research related to fungal stress biology. The Symposium was divided into seven topics: 1. Fungal biology in extreme environments; 2. Stress mechanisms and responses in fungi: molecular biology, biochemistry, biophysics, and cellular biology; 3. Fungal photobiology in the context of stress; 4. Role of stress in fungal pathogenesis; 5. Fungal stress and bioremediation; 6. Fungal stress in agriculture and forestry; and 7. Fungal stress in industrial applications. This article provides an overview of the science presented and discussed at ISFUS-2017.

Using slaughterhouse waste in a biochemical-based biorefinery - results from pilot scale tests.

A novel biorefinery concept was piloted using protein-rich slaughterhouse waste, chicken manure and straw as feedstocks. The basic idea was to provide a proof of concept for the production of platform chemicals and biofuels from organic waste materials at non-septic conditions. The desired biochemical routes were 2,3-butanediol and acetone-butanol fermentation. The results showed that hydrolysis resulted only in low amounts of easily degradable carbohydrates. However, amino acids released from the protein-rich slaughterhouse waste were utilized and fermented by the bacteria in the process. Product formation was directed towards acidogenic compounds rather than solventogenic products due to increasing pH-value affected by ammonia release during amino acid fermentation. Hence, the process was not effective for 2,3-butanediol production, whereas butyrate, propionate, γ-aminobutyrate and valerate were predominantly produced. This offered fast means for converting tedious protein-rich waste mixtures into utilizable chemical goods. Furthermore, the residual liquid from the bioreactor showed significantly higher biogas production potential than the corresponding substrates. The combination of the biorefinery approach to produce chemicals and biofuels with anaerobic digestion of the residues to recover energy in form of methane and nutrients that can be utilized for animal feed production could be a feasible concept for organic waste utilization.

Volatile fatty acids as an added value from biowaste.

The aim of the present work was to provide proof of concept of employing a co-culture of K. mobilis and E. coli for producing short and medium chain volatile fatty acids (VFAs) from kitchen biowaste and potato peels. To this aim, experiments were carried out at pilot-scale installation with a bioreactor of 250L. Different feeding strategies were tested under microaerobic conditions, at pH 6.0-6.5 in order to enhance chain elongation. Acetic acid and ethanol were dominating products in the initial stages of the bioprocess, but in a relatively short time of approx. 20-22h from the process start accumulation of propionic acid took place followed by a chain elongation to butyric and valeric acids. The highest final products yield of 325mg/g TS was achieved for the substrate load of 99.1g TS/L (VS of 91.1g/L) and pH 6.5, with the productivity of 448mg/L/h. However, the highest average VFAs chain length (3.77C) was observed in the process run with the loading of 63.2g TS/L and pH 6.0. In this study, we demonstrated that the existing symbiosis of the co-culture of K. mobilis and E. coli favours formation and chain elongation of VFA, induced most likely by the enhanced ethanol formation. Our finding differs from the previous research which focus mostly on anaerobic conditions of VFAs production. The results provide good basis for further optimisation of VFAs production process.

Proteolytic degradation and deactivation of amphibian skin peptides obtained by electrical stimulation of their dorsal glands.

Amphibians are among the oldest creatures on our planet. Their only defensive weapon efficient against microorganisms and predators involves their skin secretion. The wide range of biological activities of the peptides in the skin secretion of amphibians makes these compounds rather interesting for generation of prospective pharmaceuticals. The first step in studying these molecules requires their structures to be established. Mass spectrometry is the most powerful tool for this purpose. The sampling and sample preparation stages preceding mass spectrometry experiments appear to be rather crucial. The results obtained here demonstrate that these preparation procedures might lead to partial or complete loss of the bioactive peptides in the secretion. Five minutes in water was enough to completely destroy all of the bioactive peptides in the skin secretion of the marsh frog (Rana ridibunda); even immediate addition of methanol to the water solution of the peptides did not prevent partial destruction. Concerted effort should be directed towards development of the most efficient procedure to keep the secreted peptides intact. Graphical Abstract ᅟ.

Enhanced recovery, enrichment and detection of Mycobacterium marinum with the Portable Microbe Enrichment Unit (PMEU).

The use of PMEU significantly accelerated the growth of otherwise slowly growing Mycobacterium sp. Compared to the static reference cultures, M. marinum was detected after 24-48h of cultivation in the PMEU Spectrion(®) equipped with infrared (IR) sensors. Parallel static cultures did not exhibit or indicate mycobacterial growth within these time limits, and essentially no growth was found in them. The PMEU approach could provide a powerful tool for the rapid diagnosis and determination of environmental and clinical isolates of slow-growing species of mycobacteria. This approach also provides a method for improving diagnostics for M. tuberculosis and other pathogenic mycobacteria, including their antibiotic resistant forms, which represents a significant health problem worldwide and in Africa in particular.

Development of microbiological field methodology for water and food-chain hygiene analysis of Campylobacter spp. and Yersinia spp. in Burkina Faso, West Africa.

Field-adaptable research methods for identifying Campylobacter sp., Yersinia sp. and other pathogenic and indicator bacteria were designed in Finland and tested in Burkina Faso. Several bacterial groups were also validated from artificially contaminated samples. Campylobacter strains were cultivated using an innovative gas generation system: The 'Portable Microbe Enrichment Unit' (PMEU) which provides microaerobic gas flow into the enrichment broth. This enhanced cultivation system produced rapid growth of several isolates of campylobacteria from water and chicken samples. The latter were obtained from local marketplace samples. No yersinias were found in the field studies, whereas they were readily recovered from the spiked samples, as well as Salmonella sp. and Escherichia coli strains. The PMEU method turned out to be reliable for monitoring of water and food hygiene in remote locations.

Enhanced mycobacterial diagnostics in liquid medium by microaerobic bubble flow in Portable Microbe Enrichment Unit.

Portable Microbe Enrichment Unit (PMEU) method with microaerobic bubbling speeded up the growth of otherwise slowly starting and propagating Mycobacterium sp. Mycobacterium fortuitum growth was detected after 10-11h and Mycobacterium marinum produced clear growth in 4 days. A mycobacterial environmental isolate was verified in 2 days in the PMEU Spectrion(®) equipped with infrared sensors. In parallel static (without gas bubbling) cultures hardly any growth occurred. In conclusion, PMEU technology provided thus a rapid detection of environmental and clinical mycobacterial isolates. It would also help in the field diagnosis of antibiotic resistant Mycobacterium tuberculosis.

Fast coliform detection in portable microbe enrichment unit (PMEU) with Colilert(®) medium and bubbling.

UNLABELLED: Laboratory strains of coliforms Escherichia coli and Klebsiella mobilis were used to artificially contaminate water samples in two different cultivation and detection systems, without and with bubble flow. Samples were collected with an automated system (ASCS). The positive coliform signal caused the color change into yellow (at 550-570nm). This signal could also be transmitted on-line to cell phones. E. coli containing samples emitted UV fluorescence at 480-560nm when activated by UV light. If cultivation was started with inocula varying from 10,000 to 1cfu/ml, the positive detection was obtained between 2 and 18h, respectively, in Colilert medium using Coline PMEU device without gas bubbling. Accordingly, a single K. mobilis cell produced detectable growth in 18h. Various clinical E. coli strains were compared to each other with equal inoculum sizes, and they showed slight variations in the initiation and speed of growth. The gas bubble flow in PMEU Spectrion promoted the mixing and interaction of bacteria and indicator media and speeded the onset of growth. Carbon dioxide also accelerated bacterial growth. In the presence of vancomycin, the onset of E. coli culture growth was speeded up by the volatile outlet flow from previous cultures. In the last cultivation syringe in a series of five, the lag phase disappeared and the growth of the inoculum continued without major interruption. IN CONCLUSION: the stimulation of the cultures by the gas flow turned out to be a useful means for improving the detection of indicator bacteria. It could also be used in combination with antibiotic selection in the broth medium.

Gaseous CO2 signal initiates growth of butyric-acid-producing Clostridium butyricum in both pure culture and mixed cultures with Lactobacillus brevis.

Microbial strains produce numerous volatile substances in the anaerobic conditions of the human intestines. The availability of CO(2) is known to be a prerequisite for bacterial growth in general. In experiments with anaerobic Lactobacillus brevis and Clostridium butyricum bacteria in the Portable Microbial Enrichment Unit (PMEU) it was shown that these strains interact; this interaction being mediated by CO(2) emission. CO(2) promoted clostridial growth in pure cultures and mixed cultures with lactobacilli. The growth of C. butyricum in pure cultures was much delayed or did not start at all without CO(2) from outside. Conversely, the onset of growth was provoked by a short (15 min) CO(2) burst. In mixed cultures the presence of lactobacilli in equal numbers speeded up the onset of clostridial growth by 10 h. If C. butyricum cultures designated as PMEU 1, 2, and 3 in cultivation syringes were chained by connecting the gas flow thereby allowing the volatiles of the preceding syringe culture to bubble to the next one, the growth started in 20, 10, or 6 h, respectively. This effect of gaseous emissions from other cultures speeding up the bacterial growth initiation was abolished if the gas was passed through sodium hydroxide to remove the CO(2). The positive contribution of lactobacilli to the growth of butyric-acid-producing clostridia documented in this simulation experiment with PMEU has in vivo implications and indicates molecular communication between the species. CO(2) is a necessary signal for the growth of clostridia, and lactobacilli can promote clostridial growth in mixed cultures where both bacteria grow well with mutual benefit.

Lactic Acid bacteria enriched from human gastric biopsies.

The purpose of this paper was to check if viable bacteria, in particular lactic acid bacteria (LAB), could be enriched from biopsies obtained from healthy gastroscopy patients. Gastric biopsies were obtained from 13 gastroscopy patients and subjected to an anaerobic or microaerophilic enrichment procedure utilizing the Portable Microbe Enrichment Unit (PMEU). Profuse microbial growth was observed in most cases. Samples plated on MRS showed high numbers of LAB. The most common species characterized were Lactobacillus reuteri, Lact. salivarius, and Streptococcus salivarius. The results demonstrate a continuous presence of viable LAB in healthy stomach. The species are similar to those traditionally used in food applications. The gastric LAB strains could have a potential in developing probiotic foods aimed specially on the upper part of the gastrointestinal tract.

Enterobacterial microflora in infancy - a case study with enhanced enrichment.

OBJECTIVE: To validate PMEU (Portable Microbe Enrichment Unit) method for monitoring the composition and development of infantile intestinal enterobacterial microflora. METHODS: A case study of a boy with neonatal sepsis is presented. During the first 32 months, he was given 19 systemic antibiotic treatments representing seven different antibiotic classes. Seven fecal samples collected at ages from 3.4 to 31.6 months were studied for enterobacterial strains by a combination of enhanced enrichment culture in the PMEU and plate culture. The identification and phenotypic characterization of the isolates was performed by biochemical tests. RESULTS: 37/51 (73%) of the enterobacterial isolates were detected only after the PMEU enrichment. In most samples Escherichia coli strains were predominating, but also several Enterobacter, Klebsiella, Pantoea and Proteus strains could be isolated. It seemed that the antibiotic medications remarkably delayed the development of the intestinal microflora, because first enterobacterial strains were detected only after 6 months of age. CONCLUSIONS: The enrichment step turned out to essentially improve the characterization and monitoring of the intestinal enterobacterial microbiota of infants. Compared to plate culture the amount of isolates was 2.6-fold by the PMEU enrichment. This study gives an idea on the development and succession of microbial communities in the gastrointestinal tract and on the variation of the strains due to the intestinal environmental factors.

Growth and gaseous emissions of pure and mixed small intestinal bacterial cultures: Effects of bile and vancomycin.

Simultaneous cultivations in anaerobiosis, aerobiosis and with microaerobic gas mixture were used to clarify the bile (oxgall) effects on the pure and mixed cultures of enterobacterial strains in simulations in Portable Microbe Enrichment Unit (PMEU) linked with ChemPro100i((R)) gas detector. The effects of vancomycin were evaluated in aerobic cultures. Growth and metabolic activity of cultures were also followed by measuring sugar consumption, pH alterations, and colony counts on BD CHROMagar Orientation plates. Results showed that the two fermentatively different strains of facultative anaerobes, Escherichia coli E 17 and Klebsiella mobilis ATCC 13048 grew in balance regardless of oxygen level, bile acid concentration or other components of the mixed cultures, Bacillus cereus or Staphylococcus aureus. When the evaporations of the mixed cultures of E. coli, K. mobilis and S. aureus were compared with the emissions of the corresponding pure cultures by ChemPro100i((R)) gas sensing detector, the pure cultures of bile resistant E. coli and K. mobilis produced more gaseous components than the mixed culture indicating that these organisms cooperate and use the substrate more effectively together than separately. A survey of the aseptic bacterial isolations from the bile tract in a big University Hospital, (Salzburg, Austria) during 3 years, showed that these bacterial groups dominated. Only 13.24% of the 287 patient samples were sterile, and around 180 strains of both E. coli and Klebsiella/Enterobacter groups were found amongst 973 isolates from 249 patients (together 35.57%). Enterococcus sp. accounted for 246 isolates being the largest group of strains (24.25% of all the isolates). In anaerobiosis it was shown that Klebsiella neutralized the acids produced in the mixed acid fermentation of the E. coli. The ethanol produced from both groups evaporated in the gas stream of the PMEU culturing step and its formation also removes excess acidity from the cultures. The synergistic behaviour and symbiotic function between E. coli and Klebsiella/Enterobacter strains is suggested.

Enhanced enrichment and detection of thermotolerant Campylobacter species from water using the Portable Microbe Enrichment Unit and real-time PCR.

An enhanced enrichment using the Portable Microbe Enrichment Unit (PMEU) with the microaerobic bubbling of broths was applied for the detection of thermotolerant Campylobacter species from water. This PMEU enrichment was compared with the conventional static enrichment of the international standard ISO 17995:2005. In addition, Campylobacter detection after enrichment using a real-time PCR detection was compared with colony counts. The tests with stressed Campylobacter jejuni cells in drinking water indicated that the PMEU enrichment yielded a significantly higher number of Campylobacter cells in the Bolton broth compared with the conventional static incubation. Application of the real-time PCR technique shortened the Campylobacter detection time. This combination of method modifications can be used for Campylobacter detection from water and adds methodological repertoire for the rapid survey and management of waterborne outbreaks.

Fast detection of bacterial growth by using Portable Microbe Enrichment Unit (PMEU) and ChemPro100i((R)) gas sensor.

The combination of the Portable Microbe Enrichment Unit (PMEU) cultivation and ChemPro100i((R)) Chemical Detector helps the fast detection of hospital infections. The first alarm of the microbe metabolism and growth in the enrichment culture was achieved in 2-3h when pure cultures of strains isolated from neonatal patients were tested. For the detection of minor concentrations of the bacteria, Bacillus, Staphylococcus, Klebsiella and Escherichia strains were diluted to 0.5-600cells per ml. Their preliminary detection was achieved within 4-5h. The combination of enrichment culture (PMEU) and detection of bacterial products by ion mobility spectrometer is a rapid and sensitive method for diagnosis of bacterial growth. This finding warrants further studies with clinical samples.

Dualistic acidic and neutral glucose fermentation balance in small intestine: Simulation in vitro.

Intestinal microbes live in diminished or deprived oxygen conditions. Facultative anaerobic bacteria of the family Enterobacteriaceae ferment glucose in the gut using two main pathways: mixed acid and neutral fermentations. The aim of the present study was to clarify the roles of these fermentations in an in vitro model. Acid-producing Escherichia coli ATCC 25922 and neutral end-products producing Klebsiella mobilis ATCC 13048 were cultured in a Portable Microbe Enrichment Unit (PMEU) which permitted the selection of the gas phase in standardized conditions. In the butanediol production experiments also Klebsiella pneumoniae ssp. pneumoniae IIIa2 E111 strain was also used. Two isomers of 2,3-butanediol were observed in both aerobic and microaerobic Klebsiella cultures. During 7h cultures both E. coli and K. mobilis grew from densities of some million up to some billions per ml. The growth was almost equal in both aerobic and microaerobic conditions. In anaerobic conditions a balance prevailed in mixed E. coli and Klebsiella cultures between the species (final pH was 5.8-6.1), whereas in aerobic conditions the klebsiellas were favoured, and the pH rose up to about 8.5 (in 20h) as in pure Klebsiella cultures. In microaerobic Klebsiella cultures the final pH varied between 6.7 and 7.5. In mixed cultures the growth yields of both bacteria equalled those of the separate pure cultures indicating mutual benefits of the co-existence. Apparently E. coli produced a set of organic acids, which lowered pH. Klebsiellas increased the pH up to 2units probably due to acid conversion in anaerobic conditions to ethanol. We propose that the balance of acid and ethanol as well 2,3-butanediol fermentations contributes to the small bowel pH regulation, which also aids the host in the nutrient uptake in the small intestine.

Aerobic and anaerobic growth modes and expression of type 1 fimbriae in Salmonella.

The aim of this study was to clarify the growth rates of facultatively anaerobic Salmonella enterica serovar Enteritidis strain in aerobic and anaerobic conditions and the expression of type 1 fimbriae in relation to the growth phases. The cultivation was carried out in a Portable Microbe Enrichment Unit (PMEU) where in same conditions one can grow the cells in parallel by modifying, e.g. aerobiosis only. The results obtained show that although the anaerobic metabolism is generally believed to be a slower producer of biomass or metabolites, in these circumstances S. enterica serovar Enteritidis strain gave comparable growth rates in anaerobiosis with nitrogenation as in aerobic cultures with constant aeration. Fimbrial antigens were produced in the beginning of logarithmic phase of the growth cycle both in the aerobic and anaerobic conditions. The fimbria remained in the presence of oxygen. This capability is possibly used for the intrusion of oxygen containing tissues of host body by the invading pathogens. In conclusion S. enterica serovar Enteritidis strain suspensions grow equally well in constant nitrogenation and aeration, and fimbria were produced in both conditions, during the early logarithmic phase but they prevailed in the presence of aeration.

Semmelweis' present day follow-up: Updating bacterial sampling and enrichment in clinical hygiene.

Potentially dangerous antibiotic resistant contaminants have permanently penetrated at least well-off western populations. The danger is so evident that some hospitals have started to refuse accepting patients who carry such bacteria. Sampling and enrichment measures in hygiene monitoring must be updated as they are corner stones in handling the problems and safeguarding the health care units. Their patients, when exposed to microorganisms are strenuous to treat. Sometimes even this fails, if the infections are spreading in weakened patients. The present review summarizes currently used technologies and the abilities of bacteria to avoid detection. Improved protocols on environmental monitoring in healthcare units are required. They should be comparable with contamination control in industries. Actually these measures in health care should be even stricter because human lives are directly endangered as the resistance of especially elderly patients is low.