Iatrogenic pleural empyema caused by Propionibacterium acnes

Propionibacterium acnes is one of the commensals living on the human skin and glands, implicated mainly in acnes, but seldom in deep infection. Pleural empyema is rarely complicated with closed thoracostomy. We experienced 1 case of empyema caused by P. acnes after pleural biopsy and closed thoracostomy through a percutaneous pigtail catheter. A 79-year-old man was admitted for cough, purulent sputum and shortness of breath. Three weeks ago, closed thoracostomy and pleural biopsy were performed to confirm a diagnosis for his recurrent pleural effusion. He had increased amount of right pleural effusion. Through the pigtail catheter, pleural effusion was removed. Gram-positive rods were observed in Gram stain, but not cultured. By 16S rRNA analysis, P. acnes was confirmed as the pathogen. His empyema was repeatedly treated with antibiotics, fibrolysis and irrigation. Pleural decortication was recommended. We report the first case of empyema with P. acnes in Korea, possibly complicated with closed thoracostomy procedures.

Lactobacillus plantarum-derived Extracellular Vesicles Protect Atopic Dermatitis Induced by Staphylococcus aureus-derived Extracellular Vesicles

PURPOSE: The microbial environment is an important factor that contributes to the pathogenesis of atopic dermatitis (AD). Recently, it was revealed that not only bacteria itself but also extracellular vesicles (EVs) secreted from bacteria affect the allergic inflammation process. However, almost all research carried out so far was related to local microorganisms, not the systemic microbial distribution. We aimed to compare the bacterial EV composition between AD patients and healthy subjects and to experimentally find out the beneficial effect of some bacterial EV composition METHODS: Twenty-seven AD patients and 6 healthy control subjects were enrolled. After urine and serum were obtained, EVs were prepared from samples. Metagenomic analysis of 16s ribosomal DNA extracted from the EVs was performed, and bacteria showing the greatest difference between controls and patients were identified. In vitro and in vivo therapeutic effects of significant bacterial EV were evaluated with keratinocytes and with Staphylococcus aureus-induced mouse AD models, respectively. RESULTS: The proportions of Lactococcus, Leuconostoc and Lactobacillus EVs were significantly higher and those of Alicyclobacillus and Propionibacterium were lower in the control group than in the AD patient group. Therefore, lactic acid bacteria were considered to be important ones that contribute to the difference between the patient and control groups. In vitro, interleukin (IL)-6 from keratinocytes and macrophages decreased and cell viability was restored with Lactobacillus plantarum-derived EV treatment prior to S. aureus EV treatment. In S. aureus-induced mouse AD models, L. plantarum-derived EV administration reduced epidermal thickening and the IL-4 level. CONCLUSIONS: We suggested the protective role of lactic acid bacteria in AD based on metagenomic analysis. Experimental findings further suggest that L. plantarum-derived EV could help prevent skin inflammation.

An overview of biotechnological production of propionic acid: from upstream to downstream processes

The increasing demand for propionic acid (PA) production and its wide applications in several industries, especially the food industry (as a preservative and satiety inducer), have led to studies on the low-cost biosynthesis of this acid. This paper gives an overview of the biotechnological aspects of PA production and introduces Propionibacterium as the most popular organism for PA production. Moreover, all process variables influencing the production yield, different simple and complex carbon sources, the metabolic pathway of production, engineered mutants with increased productivity, and modified tolerance against high concentrations of acid have been described. Furthermore, possible methods of extraction and analysis of this organic acid, several applied bioreactors, and different culture systems and substrates are introduced. It can be concluded that maximum biomass and PA production may be achieved using metabolically engineered microorganisms and analyzing the most significant factors influencing yield. To date, the maximum reported yield for PA production is 0.973 g·g-1, obtained from Propionibacterium acidipropionici in a three-electrode amperometric culture system in medium containing 0.4 mM cobalt sepulchrate. In addition, the best promising substrate for PA bioproduction may be achieved using glycerol as a carbon source in an extractive continuous fermentation. Simultaneous production of PA and vitamin B12 is suggested, and finally, the limitations of and strategies for competitive microbial production with respect to chemical process from an economical point of view are proposed and presented. Finally, some future trends for bioproduction of PA are suggested.

G2A Attenuates Propionibacterium acnes Induction of Inflammatory Cytokines in Human Monocytes

BACKGROUND: Acne vulgaris is a disease of the pilosebaceous unit characterized by increased sebum production, hyperkeratinization, and immune responses to Propionibacterium acnes (PA). Here, we explore a possible mechanism by which a lipid receptor, G2A, regulates immune responses to a commensal bacterium. OBJECTIVE: To elucidate the inflammatory properties of G2A in monocytes in response to PA stimulation. Furthermore, our study sought to investigate pathways by which lipids modulate immune responses in response to PA. METHODS: Our studies focused on monocytes collected from human peripheral blood mononuclear cells, the monocytic cell line THP-1, and a lab strain of PA. Our studies involved the use of enzyme-linked immunosorbent, Western blot, reverse transcription polymerase chain reaction, small interfering RNA (siRNA), and microarray analysis of human acne lesions in the measurements of inflammatory markers. RESULTS: G2A gene expression is higher in acne lesions compared to normal skin and is inducible by the acne therapeutic, 13-cis-retinoic acid. In vitro, PA induces both the Toll-like receptor 2-dependent expression of G2A as well as the production of the G2A ligand, 9-hydroxyoctadecadienoic acid, from human monocytes. G2A gene knockdown through siRNA enhances PA stimulation of interleukin (IL)-6, IL-8, and IL-1β possibly through increased activation of the ERK1/2 MAP kinase and nuclear factor kappa B p65 pathways. CONCLUSION: G2A may play a role in quelling inflammatory cytokine response to PA, revealing G2A as a potential attenuator of inflammatory response in a disease associated with a commensal bacterium.

Influence of Chemical- and Natural-Based Lotions on Bacterial Communities in Human Forearm Skin

Purpose of this study was to evaluate the influence of a lotion on the bacterial community in the human forearm skin. The chemical- and natural-based lotions were applied on the left and right inner forearm skins, respectively, of 14 participants, who cleansed forearm skin using sterilized cotton swabs. The germs on cotton swabs were analyzed using libraries of PCR amplicons. The genetic diversity of the bacterial communities detected on the natural-based lotion-applied skin (NLS) was significantly higher than that of the bacterial communities on the chemical-based lotion-applied skin (CLS) in all participants, except two. The diversity was estimated based on operational taxonomic unit (OTU), Chao1, Shannon, and Simpson indices. Bacterial communities obtained from the CLS and NLS were phylogenetically separated into 5 and 3 monophyletic groups, respectively, based on lotion types. The taxonomic distribution of the bacterial communities, which were composed of 198 genera in 14 phyla in the CLS and NLS, respectively, was irregularly and biasedly separated into 2 groups based on the lotion types. Among the 14 phyla, Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria were found to be relatively dominant, and 15 of the 198 genera, including Methylobacterium, Propionibacterium, Pseudomonas, Staphylococcus, Streptococcus, and Bacillus were relatively dominant (>0.5%). The taxonomic distribution of dominant bacterial communities from CLS and NLS was irregularly and biasedly separated without relation to the lotion types. In conclusion, the chemical- and natural-based lotions were responsible for changing or influencing the genetic diversity, phylogenetic separation, and taxonomic distribution of skin bacterial communities.

The microbiome of the lung and its extracellular vesicles in nonsmokers, healthy smokers and COPD patients

Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory disease, and bacterial infection plays a role in its pathogenesis. Bacteria secrete nanometer-sized extracellular vesicles (EVs), which may induce more immune dysfunction and inflammation than the bacteria themselves. We hypothesized that the microbiome of lung EVs might have distinct characteristics depending on the presence of COPD and smoking status. We analyzed and compared the microbiomes of 13 nonsmokers with normal spirometry, 13 smokers with normal spirometry (healthy smokers) and 13 patients with COPD by using 16S ribosomal RNA gene sequencing of surgical lung tissue and lung EVs. Subjects were matched for age and sex in all groups and for smoking levels in the COPD and healthy smoker groups. Each group included 12 men and 1 woman with the same mean age of 65.5 years. In all groups, EVs consistently showed more operational taxonomic units (OTUs) than lung tissue. In the healthy smoker and COPD groups, EVs had a higher Shannon index and a lower Simpson index than lung tissue and this trend was more prominent in the COPD group. Principal component analysis (PCA) showed clusters based on sample type rather than participants' clinical characteristics. Stenotrophomonas, Propionibacterium and Alicyclobacillus were the most commonly found genera. Firmicutes were highly present in the EVs of the COPD group compared with other samples or groups. Our analysis of the lung microbiome revealed that the bacterial communities present in the EVs and in the COPD group possessed distinct characteristics with differences in the OTUs, diversity indexes and PCA clustering.

Microbiota bacteriana asociada al papel moneda de circulación en Colombia / Bacterial microbiota on circulating Colombian banknotes

Diariamente los seres humanos están en interacción con objetos de uso continuo, como el papel moneda, sin el conocimiento de que estos almacenan microorganismos y de que nos exponemos al contacto con potenciales patógenos. La composición de la comunidad bacteriana en un billete colombiano fue determinada mediante el secuenciamiento profundo de librerías de amplicones 16S. Se encontraron 233 géneros bacterianos; 12 de estos géneros corresponden a especies con potencial patogénico. El género más abundante fue Propionibacterium, seguido de Streptococcus, Staphylococcus Pseudomonas . Este es el primer reporte de la diversidad bacteriana que puede ser alojada en este objeto de alta circulación en Colombia. Pocos estudios en el mundo han mostrado este nivel de detalle de la microbiota en billetes de circulación y ofrece un panorama mucho más amplio de la exposición diaria a microorganismos al utilizar papel moneda en las condiciones en las que se utiliza en Colombia.

Commonly used objects such as currency paper can be colonised by bacteria and can serve as carriers of microbes. This colonisation might expose us to unnoticed pathogenic bacteria. In this study, the researchers obtained a detailed panorama of the microbes that can be carried on currency notes in Colombia by using 454 next-generation deep sequencing of 16S amplicón libraries. A total of 233 bacterial genera were detected and classified, 12 of which are potential human pathogens. The most abundant genera were Propionibacterium, Streptococcus, Staphylococcus and Pseudomonas. To date, this is the first in-depth analysis of the microbiota carried by circulating banknotes in our continent and it offers insights into daily exposure to microbes when using banknotes in Colombia.

Efficacy of Red or Infrared Light-Emitting Diodes in a Mouse Model of Propionibacterium acnes-Induced Inflammation

BACKGROUND: Laser/light-based devices may provide an alternative to conventional acne therapeutics in some patients with nonresponsive acne. OBJECTIVE: We investigated the efficacy of red or infrared light-emitting diode (LED) devices in a mouse model of Propionibacterium acnes-induced inflammation through clinical examination and histopathological and immunohistochemical studies. METHODS: A human-derived Propionibacterium acnes suspension (10(9) colony-forming units /µl) was injected into the back of an HR-1 mouse. Then, a 28.9 J/cm2 650-nm red LED or 9.3 J/cm2 830-nm infrared LED was applied to the mouse with P. acnes-induced inflammation once daily for 2 weeks. Two weeks after treatment, histological findings with hematoxylin and eosin staining and expression levels of inflammatory biomarkers (integrin α6, neutrophils, interleukin [IL]-1β, and matrix metalloproteinase [MMP]-2/9) were evaluated in tissue specimens using immunohistochemical staining. RESULTS: Mice treated with red and infrared LED showed clinical improvement in inflammatory nodules compared to mice in the control group. Red LED was much more effective than infrared LED. Epidermal hyperplasia, comedone-like cysts, and integrin α6 expression improved to a similar extent in the red and infrared LED treatment groups and control group. Neutrophil, IL-1β, MMP-2, and MMP-9 expression after treatment with red and infrared LED decreased considerably compared to expression in the control group. CONCLUSION: In a mouse model of P. acnes-induced inflammatory nodules, red and infrared LED devices may be an alternative to conventional acne therapies. In addition, a mouse model of P. acnes-induced inflammatory nodules is helpful for laboratory research of acne.

Lung Microbiome Analysis in Steroid-Naїve Asthma Patients by Using Whole Sputum / 결핵및호흡기질환

BACKGROUND: Although recent metagenomic approaches have characterized the distinguished microbial compositions in airways of asthmatics, these results did not reach a consensus due to the small sample size, non-standardization of specimens and medication status. We conducted a metagenomics approach by using terminal restriction fragment length polymorphism (T-RFLP) analysis of the induced whole sputum representing both the cellular and fluid phases in a relative large number of steroid naïve asthmatics. METHODS: Induced whole sputum samples obtained from 36 healthy subjects and 89 steroid-naїve asthma patients were analyzed through T-RFLP analysis. RESULTS: In contrast to previous reports about microbiota in the asthmatic airways, the diversity of microbial composition was not significantly different between the controls and asthma patients (p=0.937). In an analysis of similarities, the global R-value showed a statistically significant difference but a very low separation (0.148, p=0.002). The dissimilarity in the bacterial communities between groups was 28.74%, and operational taxonomic units (OTUs) contributing to this difference were as follows: OTU 789 (Lachnospiraceae), 517 (Comamonadaceae, Acetobacteraceae , and Chloroplast), 633 (Prevotella), 645 (Actinobacteria and Propionibacterium acnes), 607 (Lactobacillus buchneri, Lactobacillus otakiensis, Lactobacillus sunkii, and Rhodobacteraceae), and 661 (Acinetobacter, Pseudomonas, and Leptotrichiaceae), and they were significantly more prevalent in the sputum of asthma patients than in the sputum of the controls. CONCLUSION: Before starting anti-asthmatic treatment, the microbiota in the whole sputum of patients with asthma showed a marginal difference from the microbiota in the whole sputum of the controls.

Antimicrobial Susceptibility and Characterization of Propionibacterium acnes by Multilocus Sequence Typing and Repetitive-Sequence-Based PCR

Propionibacterium acnes, a gram-positive, anaerobic, and aerotolerant bacterium that is found frequently in the skin as part of the human microbiome causes inflammatory acne, shoulder infection, and the contamination of medical devices. The study goals were the antibiotic resistant and molecular epidemiological characterization of the P. acnes isolates in Korea. A total of 22 P. acnes isolates originated from diverse patients were obtained from three National Culture Collections for Pathogens in South Korea. The hemolytic properties and minimum inhibitory concentrations (MIC) of five antibiotics (tetracycline, doxycycline, clindamycin, erythromycin, and minocycline) were determined. Only one isolate showed high MIC values and resistance to all five antibiotics. Genotypic characterization was achieved by multilocus sequence typing (MLST) for eight loci (aroE, guaA, tly, camp2, atpD, gmk, lepA, and sodA) and repetitive-sequence-based PCR (rep-PCR) analysis using the DiversiLab kit. MLST revealed four phylogroups that were type IA₁ (27.3%), type IA₂ (18.2%), type IB (13.6%), and type II (40.9%). Rep-PCR results demonstrated three clusters that were cluster I (39.1%), cluster II (45.5%), and cluster III (13.6%). The isolates of cluster I were part of phylogroup type IA (both IA₁ and IA₂), and the isolates of cluster II belonged to phylogroup type II. All isolates of phylogroup type IB were hemolytic and belonged to cluster III. The results of rep-PCR clustering analysis showed a good correlation with those of MLST phylogroups, suggesting that rep-PCR could be an alternative method to track P. acnes subtype lineages.

Effects of Magnesium Ascorbyl Phosphate on the Expression of Inflammatory Biomarkers after Treatment of Cultured Sebocytes with Propionibacterium acnes or Ultraviolet B Radiation

No abstract available.

Effect of Vitamin D on the Expression of Inflammatory Biomarkers in Cultured Sebocytes Treated with Propionibacterium acnes or Ultraviolet B Irradiation

No abstract available.

Effect of Vitamin D on the Expression of Inflammatory Biomarkers in Cultured Sebocytes Treated with Propionibacterium acnes or Ultraviolet B Irradiation

No abstract available.

Microbial Profile of the Stomach: Comparison between Normal Mucosa and Cancer Tissue in the Same Patient

Gastric cancer is the third most common cancer and the third most frequent cause of cancer mortality in Asia. It is predicted that gastric cancer will remain an important cause of death at least during the next half century because of the increasing number of new cases in an aging population. However, little has been revealed about the role of gastric microbes and their reaction to gastric cancer. In this study, we identified differences in the microbial communities between gastric cancer and normal gastric mucosa by comparing the microbiomes of tissues from the same patients. The clustering analysis results showed different bacterial communities between normal gastric mucosa and gastric cancer. A comparison of bacterial communities at the species level revealed that Helicobacter pylori was significantly reduced in cancer tissue compared to that in normal gastric mucosa in the same patient. A comparison at the genus level showed that Propionibacterium spp., Staphylococcus spp., and Corynebacterium spp. had significantly reduced populations in cancer tissue, whereas Clostridium spp. and Prevotella spp. had significantly increased populations in cancer tissue.

Evaluation of Efficacy and Skin Irritation during Treatment of Nadifloxacin 1% Cream in Acne Vulgaris Patient / 대한피부과학회지

BACKGROUND: Nadifloxacin is a synthetic fluoroquinolone with a broad spectrum. It has been suggested to not only have antibacterial, but also anti-inflammatory actions. In addition, it is effective against antibiotic-resistant Propionibacterium acne. Upon review of the literature, however, no studies have been performed concerning the efficacy and safety in terms of irritation potential and skin barrier changes after applying nadifloxacin. OBJECTIVE: This study intended to evaluate the skin irritation and changes of the skin barrier function associated with the administration of nadifloxacin 1% cream for the treatment of acne vulgaris. Furthermore, the efficacy and safety of nadifloxacin 1% cream were also assessed. METHODS: The changes in the lesion counts, Korean Acne Grading System (KAGS) and adverse events were recorded. The biophysical profiles of the skin, including transepidermal water loss (TEWL), skin surface hydration and redness were measured by Tewameter(R), Corneometer(R) and Mexameter(R) for 3 weeks. RESULTS: At the end of the therapy, 69% of inflammatory lesions (p<0.0001) and 27% of noninflammatory lesions (p=0.26) resulting from acne were decreased from the baseline. Adverse events were reported in two patients, but they were transient and tolerable. The skin barrier function showed no changes during the treatment periods. CONCLUSION: Nadifloxacin 1% cream is effective for the treatment of acne vulgaris without the changes of skin barrier function.

[ effects of betaine on vitamin B12 production by Propionibacterium freudenreichii]

Chemical production of vitamin B12 is a complicated process. The purpose of this study, done for the first time in Iran, was to produce vitamin B12 by Propionibacterium freudenreichii and investigate the effect of adding betaine on its yield. Propionibacterium freudenreichii was added to a fermention culture medium containing filtrated soaked corn. This was followed by incubation at 30°C and, then, adding betaine at six concentrations [0, 5, 10, 15, 20 and 30 g/l]. Separation and purification were done and the presence and the amount of vitamin B[12] produced were determined by HPLC. The most effective concentration of betaine for vitamin B[12] production [318.33 Pg/ml] was 10 g/l, which had a negative effect on dry weight of the cells [22.37 g/l]. The results demonstrated that betaine could greatly stimulate vitamin B[12] biosynthesis by Propionibacterium freudenreichii and inhibit cell growth. Based on the findings of this study, betaine added to the culture medium of Propionibacterium freudenreichii at a suitable concentration could increase the yield of vitamin B[12], paving the way to a commercial, more economic method for its production

Blood collection procedures influence contamination rates in blood culture: a prospective study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Blood culture contamination is a significant adverse event. The aim of this project was to evaluate the efficacy of a strict blood collection procedure in reducing the blood culture contamination rate.</p><p><b>METHODS</b>A prospectively controlled study was performed in two different medical areas in Peking Union Medical College Hospital (PUMCH) for 16 months (from May 2006 to September 2007). In test group, a strict blood collection procedure was carried out by trained nurses with the veinpuncture sites were scrupulously disinfected with 2.5% tincture of iodine plus 70% alcohol. In control group, commonly used procedure in PUMCH was performed with 0.45% chlorhexidine acetate plus 0.2% iodine. Blood culture positive results for 4 target organisms (Coagulase-negative staphylococci, Propionibacterium acnes, Corynebacterium species and Bacillus species) were further assessed by physicians from infectious department to determine whether a sample was true positive (pathogen) or false positive (contamination).</p><p><b>RESULTS</b>Total 9321 blood culture collections were analyzed. The blood culture contamination rate in test group was significantly lower than that in control group (5/3177 (0.16%) vs. 77/6144 (1.25%); χ(2) = 13.382, P < 0.001). The most common contaminant was Coagulase-negative staphylococcus (76.83%). The average cultural time during which contaminated samples became positive was longer than that for true pathogen samples (42.0 hours vs. 13.9 hours, P = 0.041).</p><p><b>CONCLUSION</b>Using a strict blood collection procedure can significantly reduce blood culture contamination rate.</p>

Microbial profile of asymptomatic and symptomatic teeth with primary endodontic infections by pyrosequencing / 대한치과보존학회지

OBJECTIVES: The purpose of this in vivo study was to investigate the microbial diversity in symptomatic and asymptomatic canals with primary endodontic infections by using GS FLX Titanium pyrosequencing. MATERIALS AND METHODS: Sequencing was performed on 6 teeth (symptomatic, n = 3; asymptomatic, n = 3) with primary endodontic infections. Amplicons from hypervariable region of the small-subunit ribosomal RNA gene were generated by polymerized chain reaction (PCR), and sequenced by means of the GS FLX Titanium pyrosequencing. RESULTS: On average, 10,639 and 45,455 16S rRNA sequences for asymptomatic and symptomatic teeth were obtained, respectively. Based on Ribosomal Database Project Classifier analysis, pyrosequencing identified the 141 bacterial genera in 13 phyla. The vast majority of sequences belonged to one of the seven phyla: Actinobacteria, Bacteroidetes, Firmicutes, Fusobacteria, Proteobacteria, Spirochetes, and Synergistetes. In genus level, Pyramidobacter, Streptococcus, and Leptotrichia constituted about 50% of microbial profile in asymptomatic teeth, whereas Neisseria, Propionibacterium, and Tessaracoccus were frequently found in symptomatic teeth (69%). Grouping the sequences in operational taxonomic units (3%) yielded 450 and 1,997 species level phylotypes in asymptomatic and symptomatic teeth, respectively. The total bacteria counts were significantly higher in symptomatic teeth than that of asymptomatic teeth (p < 0.05). CONCLUSIONS: GS FLX Titanium pyrosequencing could reveal a previously unidentified high bacterial diversity in primary endodontic infections.

Analysis of Clinical Characteristics in Phlyctenular Keratoconjunctivitis at a Tertiary Center

PURPOSE: To report the clinical characteristics of patients with phlyctenular keratoconjunctivitis at a tertiary care center in Korea. METHODS: The authors of the present study retrospectively reviewed the medical records of 26 patients who were diagnosed with phlyctenular keratoconjunctivitis, identifying the age and sex at diagnosis, diagnosis of the referring clinic, laterality and location of the lesions, the results of bacterial cultures from the meibum, and treatment options. RESULTS: Eighteen patients (76.2%) were female, and the mean age at diagnosis was 18.4 +/- 10.5 years. Nine patients (34.6%) showed evidence of bilateral diseases, and corneal lesions were observed in 27 eyes of 21 patients. Bacterial cultures from the meibum were performed for 17 patients, with the following results: 6 cases of Staphylococcus epidermidis, 4 cases of Corynebacterium species, and 1 case of Propionibacterium acne. Neovascularization with corneal phlyctenules occurred from the inferior quadrant mostly. Twenty-four patients received the remission treatment of prednisolone acetate 1% eye drops and evidence of regression of new vessels was observed in all patients. Three patients (16.7%) experienced recurrence despite the maintenance treatment of cyclosporine 0.05% eye drops. Visual acuities improved significantly after the remission treatment, as compared to the initial visit. CONCLUSIONS: The phlyctenular keratoconjunctivitis patients who visited the tertiary center in Korea showed similar findings regarding demographics, involvement of blepharitis, and identified microorganisms, as compared with patients in previous reports. However, corneal lesions involving the visual axis were frequent, suggesting that with proper diagnosis and treatment, improvement in visual acuities is possible.

Trials for improving the efficiency of inactivated Newcastle disease virus vaccine by using propioni-bacterium and lipopolysaccharide [IM-104] as immunostimulant

The ability to improve the efficacy of an inactivated Newcastle disease vaccine by addition of immunomodulator as propionibacterium and lipopolysaccharide [TM-104] was studied. Chickens were divided into three groups, one received inactivated Newcastle vaccine containing the IM-104, second group received vaccine alone and third group of chickens was left without vaccination as control. Blood samples were collected 3 days post vaccination, and weekly till four weeks for estimation of the effect of [IM-104] on Tlymphocyte transformation [cellular immune response]. Sero-conversion for sera at weekly intervals was done to evaluate the immune response using HI test and ELISA. Challenge test was carried out on 3[rd] and 6[th] week post vaccination for evaluation of protective efficacy of these vaccines. The results obtained revealed that [TM-104] that contain 2 extraction of E. coli lipopolysaccharide and propioni-bacterium plays roles in stimulating both T-lymphocyte and macrophage to release different types of interleukin and consequently released immunoglobulins. Also, stimulate non-specific resistance to viral infection. The results obtained by challenge test run in parallel with that of serological test