Effects of Spirulina platensis and/or Allium sativum on Antioxidant Status, Immune Response, Gut Morphology, and Intestinal Lactobacilli and Coliforms of Heat-Stressed Broiler Chicken.

This study aims to evaluate the effectiveness of the dietary addition of Spirulina platensis (SP) and/or garlic powder (GP) on heat-stressed broiler chickens. For this purpose, 600 Ross-308 broiler chicks were allocated at 22 days of age into five groups (G1-G5), each comprising six groups of 20 birds each. Chickens kept in G1 (negative control) were fed a basal diet and raised at 26 ± 1 °C. Chickens kept in G2 to G5 were exposed to periodic heat stress (35 ± 1 °C for 9 h/day) from 22 to 35 days old. Chickens in G2 (positive control) were provided a basal diet, while G3, G4, and G5 were fed a basal diet enriched with SP (1 g/kg diet), GP (200 mg/kg diet), or SP/GP (1 g SP/kg + 200 mg GP/kg diet), respectively. The assessment parameters included the chickens' performance, malondialdehyde and total antioxidant capacity, blood biochemistry, intestinal morphology, and modulation of lactobacilli and total coliforms in the intestinal microbiota. Our findings demonstrated that supplementing heat-stressed chickens with SP and/or GP significantly mitigated the negative effects on the European production efficiency index (EPEF), survival rate, cholesterol profile, and oxidative stress markers. Chickens supplemented with GP and/or SP exhibited significantly better EPEF and survivability rates. Heat stress had a significant impact on both the gut structure and gut microbiota. However, SP and/or GP supplementation improved the gut morphology, significantly increased the intestinal lactobacilli, and reduced the coliform contents. It was also found that the simultaneous feeding of SP and GP led to even higher recovery levels with improved lipid metabolites, immunity, and oxidative status. Overall, supplementing chickens with SP and/or GP can alleviate the negative effects of heat stress.

Alternative to antibiotic growth promoters: beneficial effects of Saccharomyces cerevisiae and/or Lactobacillus acidophilus supplementation on the growth performance and sustainability of broilers' production.

Although antibiotics growth promoters (AGPs), including zinc-bacitracin (ZnB), can threaten human health due to developing antimicrobial resistance, as well as drug residue in animal and poultry products, ZnB is still widely used, particularly in developing countries, for the sustainability of poultry farming. The present investigation aims to assess the use of Saccharomyces cerevisiae and Lactobacillus acidophilus, with or without a prebiotic (mannooligosaccharide, MOS), as alternatives to ZnB. For this reason, 150 one-day-old chicks were grouped into six groups, designated negative control, LA, SC, ZnB, SA + MOS, and LA + MOS (5 replicates of 5 chicks for each group). Chicks kept in the control group were fed the basal diet. Chickens kept in LA and SC groups received L. acidophilus, S. cerevisiae at a 1 g/kg diet and 2 g/Kg, respectively. Chickens kept in ZnB received ZnB at 0.5 g/kg. Chicks kept in the SC + MOS and LA + MOS were fed a basal diet containing 2 g S. cerevisiae + 1 g MOS/kg or 1 g L. acidophilus + 1 g MOS /kg, respectively. The efficacy was assessed based on the growth performance, carcass traits, meat quality, nutrient digestibility, and blood biochemistry composition during the entire trial 1-36 days of age. Results showed that chicks kept in the SC group had greater BW than the control (p < 0.05). Chicks kept in the SC, LA, SC + MOS, and LA + MOS consumed less feed than the control and Zn-B groups (p < 0.05). Supplementation with S. cerevisiae resulted in a better (p < 0.05) feed conversion rate (FCR) than the control group. Supplementation with L. acidophilus + MOS significantly increased (p < 0.05) the relative liver weight compared to those supplemented with ZnB, S. cerevisiae, and L. acidophilus. In addition, supplementation with ZnB-induced spleen hypertrophy compared to S. cerevisiae and L. acidophilus-supplemented groups (p < 0.05). Plasma, meat, and liver cholesterol, as well as the cholesterol-to-lipid ratio of meat and liver, were significantly decreased (p < 0.05) in both SC and LA groups compared to the control group. Our research indicates that adding 2 g/kg of S. cerevisiae to broiler feed can effectively replace ZnB and enhance productive performance and economic profits, making it a viable and sustainable option for broiler farming.

An overview of the use of bacteriophages in the poultry industry: Successes, challenges, and possibilities for overcoming breakdowns.

The primary contaminants in poultry are Salmonella enterica, Campylobacter jejuni, Escherichia coli, and Staphylococcus aureus. Their pathogenicity together with the widespread of these bacteria, contributes to many economic losses and poses a threat to public health. With the increasing prevalence of bacterial pathogens being resistant to most conventional antibiotics, scientists have rekindled interest in using bacteriophages as antimicrobial agents. Bacteriophage treatments have also been investigated as an alternative to antibiotics in the poultry industry. Bacteriophages' high specificity may allow them only to target a specific bacterial pathogen in the infected animal. However, a tailor-made sophisticated cocktail of different bacteriophages could broaden their antibacterial activity in typical situations with multiple clinical strains infections. Bacteriophages may not only be used in terms of reducing bacterial contamination in animals but also, under industrial conditions, they can be used as safe disinfectants to reduce contamination on food-contact surfaces or poultry carcasses. Nevertheless, bacteriophage therapies have not been developed sufficiently for widespread use. Problems with resistance, safety, specificity, and long-term stability must be addressed in particular. This review highlights the benefits, challenges, and current limitations of bacteriophage applications in the poultry industry.

Anti-Inflammatory and Antioxidative Phytogenic Substances against Secret Killers in Poultry: Current Status and Prospects.

Chronic stress is recognized as a secret killer in poultry. It is associated with systemic inflammation due to cytokine release, dysbiosis, and the so-called leaky gut syndrome, which mainly results from oxidative stress reactions that damage the barrier function of the cells lining the gut wall. Poultry, especially the genetically selected broiler breeds, frequently suffer from these chronic stress symptoms when exposed to multiple stressors in their growing environments. Since oxidative stress reactions and inflammatory damages are multi-stage and long-term processes, overshooting immune reactions and their down-stream effects also negatively affect the animal's microbiota, and finally impair its performance and commercial value. Means to counteract oxidative stress in poultry and other animals are, therefore, highly welcome. Many phytogenic substances, including flavonoids and phenolic compounds, are known to exert anti-inflammatory and antioxidant effects. In this review, firstly, the main stressors in poultry, such as heat stress, mycotoxins, dysbiosis and diets that contain oxidized lipids that trigger oxidative stress and inflammation, are discussed, along with the key transcription factors involved in the related signal transduction pathways. Secondly, the most promising phytogenic substances and their current applications to ameliorate oxidative stress and inflammation in poultry are highlighted.

Influence of COVID-19 on the sustainability of livestock performance and welfare on a global scale.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is currently spreading worldwide. The pandemic has already had significant adverse effects on human civilization, the environment, and the ecosystem at national and global levels. Moreover, the various sectors of the food production chain, particularly agriculture and livestock, have also been significantly affected in terms of production sustainability and economic losses. The global pandemic has already resulted in a sharp drop in meat, milk, and egg production. Restrictions of movement at national and international levels, implemented as a part of control strategies by public health sectors, have negatively impacted business related to the supply of raw materials for livestock farmers and farm outputs, veterinary services, farmworkers, and animal welfare. This review highlights the significant impacts of COVID-19 on the sustainability of livestock performance, welfare on a global scale, and strategies for mitigating these adverse effects.

Poultry Production and Sustainability in Developing Countries under the COVID-19 Crisis: Lessons Learned.

Poultry farming is a significant source of revenue generation for small farmers in developing countries. It plays a vital role in fulfilling the daily protein requirements of humans through meat and eggs consumption. The recently emerged pandemic Coronavirus Disease-19 (COVID-19) impacts the poultry production sector. Although the whole world is affected, these impacts may be more severe in developing countries due to their dependency on exporting necessary supplies such as feed, vaccines, drugs, and utensils. In this review, we have discussed poultry production in developing countries under the COVID-19 crisis and measures to regain the loss in the poultry industries. Generally, due to the lockdown, trade limitations have negatively impacted poultry industries, which might exacerbate global poverty. Coordinated activities have to be taken at the private and government levels to arrange soft loans so that these farms can restore their production and marketing to normal levels. In addition, here, we have focused on the supply of farm input, feed, other raw materials, management system, improved breeding efficiency, veterinary services, and marketing of egg and meat, which have to be ensured to secure a sustainable poultry production chain.

Algae and Their Metabolites as Potential Bio-Pesticides.

An increasing human population necessitates more food production, yet current techniques in agriculture, such as chemical pesticide use, have negative impacts on the ecosystems and strong public opposition. Alternatives to synthetic pesticides should be safe for humans, the environment, and be sustainable. Extremely diverse ecological niches and millions of years of competition have shaped the genomes of algae to produce a myriad of substances that may serve humans in various biotechnological areas. Among the thousands of described algal species, only a small number have been investigated for valuable metabolites, yet these revealed the potential of algal metabolites as bio-pesticides. This review focuses on macroalgae and microalgae (including cyanobacteria) and their extracts or purified compounds, that have proven to be effective antibacterial, antiviral, antifungal, nematocides, insecticides, herbicides, and plant growth stimulants. Moreover, the mechanisms of action of the majority of these metabolites against plant pests are thoroughly discussed. The available information demonstrated herbicidal activities via inhibition of photosynthesis, antimicrobial activities via induction of plant defense responses, inhibition of quorum sensing and blocking virus entry, and insecticidal activities via neurotoxicity. The discovery of antimetabolites also seems to hold great potential as one recent example showed antimicrobial and herbicidal properties. Algae, especially microalgae, represent a vast untapped resource for discovering novel and safe biopesticide compounds.

Probiotics, Prebiotics, and Phytogenic Substances for Optimizing Gut Health in Poultry.

The gut microbiota has been designated as a hidden metabolic 'organ' because of its enormous impact on host metabolism, physiology, nutrition, and immune function. The connection between the intestinal microbiota and their respective host animals is dynamic and, in general, mutually beneficial. This complicated interaction is seen as a determinant of health and disease; thus, intestinal dysbiosis is linked with several metabolic diseases. Therefore, tractable strategies targeting the regulation of intestinal microbiota can control several diseases that are closely related to inflammatory and metabolic disorders. As a result, animal health and performance are improved. One of these strategies is related to dietary supplementation with prebiotics, probiotics, and phytogenic substances. These supplements exert their effects indirectly through manipulation of gut microbiota quality and improvement in intestinal epithelial barrier. Several phytogenic substances, such as berberine, resveratrol, curcumin, carvacrol, thymol, isoflavones and hydrolyzed fibers, have been identified as potential supplements that may also act as welcome means to reduce the usage of antibiotics in feedstock, including poultry farming, through manipulation of the gut microbiome. In addition, these compounds may improve the integrity of tight junctions by controlling tight junction-related proteins and inflammatory signaling pathways in the host animals. In this review, we discuss the role of probiotics, prebiotics, and phytogenic substances in optimizing gut function in poultry.

Status and Prospects of Botanical Biopesticides in Europe and Mediterranean Countries.

Concerning human and environmental health, safe alternatives to synthetic pesticides are urgently needed. Many of the currently used synthetic pesticides are not authorized for application in organic agriculture. In addition, the developed resistances of various pests against classical pesticides necessitate the urgent demand for efficient and safe products with novel modes of action. Botanical pesticides are assumed to be effective against various crop pests, and they are easily biodegradable and available in high quantities and at a reasonable cost. Many of them may act by diverse yet unexplored mechanisms of action. It is therefore surprising that only few plant species have been developed for commercial usage as biopesticides. This article reviews the status of botanical pesticides, especially in Europe and Mediterranean countries, deepening their active principles and mechanisms of action. Moreover, some constraints and challenges in the development of novel biopesticides are highlighted.

Diversity of Coronaviruses with Particular Attention to the Interspecies Transmission of SARS-CoV-2.

In December 2019, the outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was reported in China with serious impacts on global health and economy that is still ongoing. Although interspecies transmission of coronaviruses is common and well documented, each coronavirus has a narrowly restricted host range. Coronaviruses utilize different receptors to mediate membrane fusion and replication in the cell cytoplasm. The interplay between the receptor-binding domain (RBD) of coronaviruses and their coevolution are determinants for host susceptibility. The recently emerged SARS-CoV-2 caused the coronavirus disease 2019 (COVID-19) pandemic and has also been reported in domestic and wild animals, raising the question about the responsibility of animals in virus evolution. Additionally, the COVID-19 pandemic might also substantially have an impact on animal production for a long time. In the present review, we discussed the diversity of coronaviruses in animals and thus the diversity of their receptors. Moreover, the determinants of the susceptibility of SARS-CoV-2 in several animals, with special reference to the current evidence of SARS-CoV-2 in animals, were highlighted. Finally, we shed light on the urgent demand for the implementation of the One Health concept as a collaborative global approach to mitigate the threat for both humans and animals.

Poult Enteritis and Mortality Syndrome in Turkey Poults: Causes, Diagnosis and Preventive Measures.

Poult enteritis and mortality syndrome (PEMS) is one of the most significant problem affecting turkeys and continues to cause severe economic losses worldwide. Although the specific causes of PEMS remains unknown, this syndrome might involve an interaction between several causative agents such as enteropathogenic viruses (coronaviruses, rotavirus, astroviruses and adenoviruses) and bacteria and protozoa. Non-infectious causes such as feed and management are also interconnected factors. However, it is difficult to determine the specific cause of enteric disorders under field conditions. Additionally, similarities of clinical signs and lesions hamper the accurate diagnosis. The purpose of the present review is to discuss in detail the main viral possible causative agents of PEMS and challenges in diagnosis and control.

Evolutionary insights into the furin cleavage sites of SARS-CoV-2 variants from humans and animals.

The SARS-CoV-2 spike protein Q677P/H mutation and furin cleavage site (FCS) have been shown to affect cell tropism and virus transmissibility. Here, we analyzed the frequency of Q677P/H and FCS point mutations in 1,144,793 human and 1042 animal spike protein sequences and from those of the emergent variants B.1.1.7, B.1.351, P.1, B.1.429 + B.1.427, and B.1.525, which were deposited in the database of the GISAID Initiative. Different genetic polymorphisms, particularly P681H and A688V, were detected in the FCS, mainly in human isolates, and otherwise, only pangolin and bat sequences had these mutations. Multiple FCS amino acid deletions such as Δ680SPRRA684 and Δ685RSVA688 were only detected in eight and four human isolates, respectively. Surprisingly, deletion of the entire FCS motif as Δ680SPRRARSVA688 and Δ680SPRRARSVAS689 was detected only in three human isolates. On the other hand, analysis of FCS from emergent variants showed no deletions in the FCS except for spike P681del, which was detected in seven B.1.1.7 isolates from the USA. Spike Q677P was detected only once in variant, B.1.1.7, whereas Q677H was detected in all variants, i.e., B.1.1.7 (n = 1938), B.1.351 (n = 28), P.1 (n = 9), B.1.429 + B.1.427 (n = 132), and B.1.525 (n = 1584). Structural modeling predicted that mutations or deletions at or near the FCS significantly alter the cleavage loop structure and would presumably affect furin binding. Taken together, our results show that Q677H and FCS point mutations are prevalent and may have various biological effects on the circulating variants. Therefore, we recommend urgent monitoring and surveillance of the investigated mutations, as well as laboratory assessment of their pathogenicity and transmissibility.

Characterization of Sunflower Oil Extracts from the Lichen Usnea barbata.

The increasing global emergence of multidrug resistant (MDR) pathogens is categorized as one of the most important health problems. Therefore, the discovery of novel antimicrobials is of the utmost importance. Lichens provide a rich source of natural products including unique polyketides and polyphenols. Many of them display pharmaceutical benefits. The aim of this study was directed towards the characterization of sunflower oil extracts from the fruticose lichen, Usnea barbata. The concentration of the major polyketide, usnic acid, was 1.6 mg/mL extract as determined by NMR analysis of the crude mixture corresponding to 80 mg per g of the dried lichen. The total phenolics and flavonoids were determined by photometric assays as 4.4 mg/mL (gallic acid equivalent) and 0.27 mg/mL (rutin equivalent) corresponding to 220 mg/g and 13.7 mg/g lichen, respectively. Gram-positive (e.g., Enterococcus faecalis) and Gram-negative bacteria, as well as clinical isolates of infected chickens were sensitive against these extracts as determined by agar diffusion tests. Most of these activities increased in the presence of zinc salts. The data suggest the potential usage of U. barbata extracts as natural additives and mild antibiotics in animal husbandry, especially against enterococcosis in poultry.

Development of Salmonella Enteritidis vaccine candidate based on streptomycin independent suppressor and metabolic drift rifampicin resistance-attenuating markers.

Salmonella is one of the most frequent food-borne pathogens and remains public health threat globally. The control of Salmonella in poultry, the main reservoir of non-typhoidal salmonellae, is a fundamental approach to ensure the safety of poultry products for human consumption. In the present study, a new live attenuated Salmonella enterica serovar Enteritidis vaccine candidate containing three attenuating markers based on streptomycin-independent (Sm-id) suppressor, and metabolic drift antibiotic resistance (MD- "res") was developed. The streptomycin dependent (Smd) mutants were derived from Salmonella Enteritidis wild-type strain using streptomycin. Then the Sm-id mutants were derived from the isolated Smd mutants and designated "Smd→Sm-id". A third MD- "res" marker was generated from Smd→Sm-id using rifampicin (Rif) and designated "Smd→Sm-id→Rif". The colony sizes of these mutants were stable after more than 50 serial passages on blood agar; reversion to virulence can be almost excluded. The safety and efficacy of Smd→Sm-id and Smd→Sm-id→Rif were evaluated in one-day-old commercial layer chicks. Both mutants proved to be safe in terms of clinical signs, mortalities, lesion scores of visceral organs and rapid clearance when administered orally at a dose of 108 colony forming unit (CFU), whereas birds inoculated with 108 CFU Salmonella Enteritidis wild-type strain showed diarrhea, mortalities (3/40) and necrosis in liver and spleen. Chickens vaccinated with the developed mutants showed no seroconversion; however, wild-type strain induced a significant seroconversion at 3-week-postvaccination (wpv). The developed mutants protected chickens against challenge with 108 CFU of Salmonella Enteritidis wild-type strain at 3-wpv. Vaccinated birds showed neither clinical signs nor mortalities during two-week post-challenge. In addition, the challenge strain could not be detected in pooled liver and spleen samples (0/5) at 7th day post-inoculation (dpi). However, non-vaccinated challenged birds showed diarrhea and the challenge strain was re-isolated from pooled liver and spleen samples (3/5) at 7th dpi. In conclusion, the developed mutants are safe and fully protected immunized chickens following heterologous challenge. It is obvious that the genetic characterization of these mutants and evaluation of different vaccination regimes are still in demand.

Immunization of turkeys with a DNA vaccine expressing the haemagglutinin gene of low pathogenic avian influenza virus subtype H9N2.

Low pathogenic avian influenza H9N2 is still circulating in the Middle East causing respiratory manifestations and severe economic losses in poultry. In the present study, an H9 plasmid-based DNA vaccine targeting the HA gene of H9N2 A/CK/Egypt/SCU8/2014 was developed and evaluated in turkeys. The full length of HA was cloned into vector plasmids under the control of a cytomegalovirus promoter. The in-vitro expression of the recombinant HA was demonstrated in HeLa cells transfected with the plasmids pVAX1-H9 or pCR-H9 using western blot and Immunofluorescent assay (IFA). The efficacy of pVAX-H9 and pCR- H9, naked or saponin-adjuvanted, was evaluated in turkey poults at 3 weeks and challenged with A/CK/Egypt/SCU8/2014 (106 EID50/bird at 3 weeks post-vaccination. The efficacy was assesses based on virus shedding, oropharyngeal and cloacal, as well as seroconversion using haemagglutination inhibition (HI) test. All immunized birds showed high HI antibody titers (7-8 log2) at 3 weeks post-vaccination. None of the birds vaccinated with naked or saponin-adjuvanted pVAX-H9 or pCR-H9 showed any clinical signs. The pVAX-H9 and pCR-H9 alone did not prevent cloacal and oropharyngeal virus shedding, however, saponin-adjuvanted pVAX1-H9 and pCR-H9 prevented cloacal and oropharyngeal virus shedding at 3 and 5 days post challenge, respectively. In conclusion, DNA vaccination with pVAX1-H9 and pCR-H9 could protect turkey from the H9N2 virus, but vaccination regimes need to be improved.

Mutational spectra of SARS-CoV-2 isolated from animals.

Coronaviruses are ubiquitous and infect a wide spectrum of animals and humans. The newly emerged severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has become a worldwide pandemic. To address the role that animals may play in the evolution of SARS-CoV-2, the full genome sequences of SARS-CoV-2 isolated from animals were compared with SARS-CoV-2 human isolates from the same clade and geographic region. Phylogenetic analysis of SARS-CoV-2 isolated from the cat, dog, mink, mouse, and tiger revealed a close relationship with SARS-CoV-2 human isolates from the same clade and geographic region with sequence identities of 99.94-99.99%. The deduced amino acid sequence of spike (S) protein revealed the presence of a furin cleavage site (682RRAR▾685), which did not differ among all SARS-CoV-2 isolates from animals and humans. SARS-CoV-2 isolates from minks exhibited two amino acid substitutions (G261D, A262S) in the N-terminal domain of S protein and four (L452M, Y453F, F486L, N501T) in the receptor-binding motif (RBM). In the mouse, the S protein had two amino acid substitutions, one in the RBM (Q498H) and the other (N969S) in the heptad repeat 1. SARS-CoV-2 isolated from minks furtherly exhibited three unique amino acid substitutions in the nucleocapsid (N)protein. In the cat, two unique amino acid substitutions were discovered in the N (T247I) and matrix (T175M) proteins. Additionally, SARS-CoV-2 isolated from minks possessed sixteen, four, and two unique amino acid substitutions in the open reading frame 1ab (ORF1ab), ORF3a, and ORF6, respectively. Dog and cat SARS-CoV-2 isolates showed one and seven unique amino acid substitutions in ORF1ab, respectively. Further studies may be necessary to determine the pathogenic significance of these amino acid substitutions to understand the molecular epidemiology and evolution of SARS-CoV-2.

Isolation and characterization of Salmonella Enteritidis and Salmonella Typhimurium from chicken meat in Egypt.

INTRODUCTION: Salmonella enterica serovars Enteritidis and Typhimurium represent the major serovars associated with human salmonellosis. Contamination of meat products with these serovars is considered the main source of infection. METHODOLOGY: In this study, 100 raw chicken meat samples were investigated for the presence of Salmonella spp., which were subsequently identified based on biochemical and serological tests as well as matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) profile. Furthermore, the isolated serovars were examined using multiplex polymerase chain reaction (PCR) for the presence of virulence genes suspected to have a role in infection. RESULTS: S. Enteritidis was isolated from two samples (2%), while S. Typhimurium was isolated from three samples (3%) of chicken meat. Of the 17 examined virulence genes using multiplex PCR, the sitC, sopB, sifA, lpfC, spaN, sipB, invA, spiA, and msgA genes were detected in S. Enteritidis. However, the sitC, iroN, sopB, sifA, lpfC, spaN, sipB, invA, and tolC genes were successfully amplified in S. Typhimurium. CONCLUSIONS: The detection of S. Enteritidis and S. Typhimurium in meat, even at low incidence, has important implications. In addition, the data presented here is the first attempt to identify a wide range of virulence genes in Egyptian Salmonella isolates recovered from meat products. A strict public health and food safety regime is urgently needed in order to decrease the human health hazard risk associated with salmonellosis.

Phenotypic and Genotypic Characterization of Bacteriocinogenic Enterococci Against Clostridium botulinum.

The present study aimed to characterize Enterococcus faecalis (n = -6) and Enterococcus faecium (n = 1) isolated from healthy chickens to find a novel perspective probiotic candidate that antagonize Clostridium botulinum types A, B, D, and E. The isolated enterococci were characterized based on phenotypic properties, PCR, and matrix-assisted laser desorption/ionization time of flight (MALDI-TOF). The virulence determinants including hemolytic activity on blood agar, gelatinase activity, sensitivity to vancomycin, and presence of gelatinase (gelE) and enterococcal surface protein (esp) virulence genes were investigated. Also, the presence of enterocin structural genes enterocin A, enterocin B, enterocin P, enterocin L50A/B, bacteriocin 31, enterocin AS48, enterocin 1071A/1071B, and enterocin 96 were assessed using PCR. Lastly, the antagonistic effect of the selected Enterococcus spp. on the growth of C. botulinum types A, B, D, and E was studied. The obtained results showed that four out of six E. faecalis and one E. faecium proved to be free from the tested virulence markers. All tested enterococci strains exhibited more than one of the tested enterocin. Interestingly, E. faecalis and E. faecium significantly restrained the growth of C. botulinum types A, B, D, and E. In conclusion, although, the data presented showed that bacteriocinogenic Enterococcus strains lacking of virulence determinants could be potentially used as a probiotic candidate against C. botulinum in vitro; however, further investigations are still urgently required to verify the beneficial effects of the tested Enterococcus spp. in vivo.

The influence of polyunsaturated fatty acids on the phospholipase D isoforms trafficking and activity in mast cells.

The impact of polyunsaturated fatty acid (PUFA) supplementation on phospholipase D (PLD) trafficking and activity in mast cells was investigated. The enrichment of mast cells with different PUFA including α-linolenic acid (LNA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), linoleic acid (LA) or arachidonic acid (AA) revealed a PUFA-mediated modulation of the mastoparan-stimulated PLD trafficking and activity. All PUFA examined, except AA, prevented the migration of the PLD1 to the plasma membrane. For PLD2 no PUFA effects on trafficking could be observed. Moreover, PUFA supplementation resulted in an increase of mastoparan-stimulated total PLD activity, which correlated with the number of double bonds of the supplemented fatty acids. To investigate, which PLD isoform was affected by PUFA, stimulated mast cells were supplemented with DHA or AA in the presence of specific PLD-isoform inhibitors. It was found that both DHA and AA diminished the inhibition of PLD activity in the presence of a PLD1 inhibitor. By contrast, only AA diminished the inhibition of PLD activity in the presence of a PLD2 inhibitor. Thus, PUFA modulate the trafficking and activity of PLD isoforms in mast cells differently. This may, in part, account for the immunomodulatory effect of unsaturated fatty acids and contributes to our understanding of the modulation of mast cell activity by PUFA.

Polyunsaturated fatty acid supplements modulate mast cell membrane microdomain composition.

In the present study, the lipid raft composition of a canine mastocytoma cell line (C2) was analyzed. Lipid rafts were well separated from non-raft plasma membranes using a detergent-free isolation technique. To study the influence of n-3 and n-6 polyunsaturated fatty acids (PUFA) on raft fatty acid composition in comparison to non-raft cell membrane, C2 were supplemented with one of the following: α-linolenic acid, eicosapentaenoic acid, docosahexaenoic acid, linoleic acid or arachidonic acid. Enrichment of the culture medium with a specific PUFA resulted in an increase in the content of this fatty acid both in rafts and non-raft membranes. Contents of cholesterol and protein were found not to be affected by the changes in the fatty acid profiles. In conclusion, our data provide strong evidence that PUFA modulate lipid composition and physiological properties of membrane micro domains of mast cells which in turn may have effects on mast cell function.