Jakeen K. El-Jakee

Emerging of coagulase negative staphylococci as a cause of mastitis in dairy animals: An environmental hazard ☆

Abstract In Egypt, knowledge about the coagulase negative staphylococci (CNS) involved in mastitic animals is limited. CNS have emerged to be pathogens causing intramammary infections in Egyptian dairy herds. Therefore, the current study was conducted to investigate the occurrence of CNS in dairy ruminants (cattle, buffaloes, sheep and goats). A total of 884 quarter milk samples were investigated to study the prevalence of CNS among mastitic and subclinically mastitic cows, buffalo–cows, ewes and does in Egypt. Identification of the isolates was achieved using API staph test and polymerase chain reaction (PCR). CNS were isolated from the examined subclinical mastitic cattle, buffaloes, sheep and goats with percentages of 16.6%, 59.4%, 50% and 55.6%, respectively. Staphylococcus xylosus, Staphylococcus cohnii, Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus saprophyticus, Staphylococcus chromogenes, Staphylococcus lentus, Staphylococcus lugdunensis and Staphylococcus simulans were identified as CNS that recovered from the examined milk samples. The CNS as mastitis-causing agents could not be neglected as they can cause substantial economic losses.

Molecular characterization of Escherichia coli O157:H7 recovered from meat and meat products relevant to human health in Riyadh, Saudi Arabia.

Raw meat can harbor pathogenic bacteria, potentially harmful to humans such as Escherichia coli O157:H7 causing diarrhea and hemolytic-uremic syndrome (HS). Therefore, the current study was carried out to evaluate the prevalence and the molecular detection characterization of E. coli serotype O157:H7 recovered from raw meat and meat products collected from Saudi Arabia. During the period of 25th January 2013 to 25th March 2014, 370 meat samples were collected from abattoirs and markets located in Riyadh, Saudi Arabia “200 raw meat samples and 170 meat products”. Bacteriological analysis of the meat samples and serotyping of the isolated E. coli revealed the isolation of 11 (2.97%) strains of E. coli O157:H7. Isolation of E. coli O157:H7 in raw beef, chicken and mutton were 2%, 2.5%, and 2.5%, respectively, however, there was no occurrence in raw turkey. The incidences of E. coli O157:H7 in ground beef, beef burgers, beef sausage, ground chicken and chicken burgers were 5%, 10%, 0.0%, 5% and 0.0%, respectively. The multiplex PCR assay revealed that 3 (27.27%) out of 11 E. coli O157:H7 isolates from raw beef, chicken and mutton had stx1, stx2, and eae while 5 (45.45%) E. coli O157:H7 isolates from ground beef, ground chicken, and raw beef had both stx1 and stx2. However, from beef burgers, only one E. coli O157:H7 isolate had stx1 while two were positive for hlyA gene. These results call for urgent attention toward appropriate controls and good hygienic practices in dealing with raw meat.

Comparative studies for serodiagnosis of haemorrhagic septicaemia in cattle sera.

Haemorrhagic septicaemia caused by Pasteurella multocida is a major epizootic disease in cattle and buffaloes in developing countries with high morbidity and mortality rate. In the present study, a total of 88 P. multocida isolates were isolated from 256 nasopharyngeal swabs and lung tissues samples (34.4%) during the period from January, 2013 to March, 2014 from different governorates located in Egypt. Dead calves showed the highest percentage of P. multocida isolation followed by the emergency slaughtered calves, diseased calves then apparently healthy ones. These isolates were confirmed as P. multocida microscopically, biochemically by traditional tests and by API 20E commercial kit then by PCR. The percentages of positive serum samples using somatic antigen and micro-agglutination test at 1/1280 diluted serum were 10%, 54.49% and 0% in apparently healthy, diseased and emergency slaughtered samples, respectively whereas, the percentages using capsular antigen and indirect haemagglutination test were 40%, 60.89% and 60% in apparently healthy, diseased and emergency slaughtered samples, respectively. The ELISA showed the highest sensitivity for diagnosing P. multocida in apparently healthy, diseased and emergency slaughtered animals with percentages of 42%; 92.9% and 80%, respectively. The obtained results revealed that the ELISA using capsular antigen of P. multocida is a more sensitive and specific serological test for diagnosis of haemorrhagic septicaemia.

Molecular and serotyping characterization of shiga toxogenic Escherichia coli associated with food collected from Saudi Arabia.

Shiga toxin-producing Escherichia coli (STEC) strains are considered as one of the major food-borne disease agents in humans worldwide. STEC strains, also called verotoxin-producing E. coli strains. The objectives of the present study were serotyping and molecular characterization of shiga toxigenic E. coli associated with raw meat and milk samples collected from Riyadh, Saudi Arabia. A total of 540 milk samples were collected from 5 dairy farms and 150 raw meat samples were collected from different abattoirs located in Riyadh, Saudi Arabia. E. coli were recovered from 86 milk samples (15.93%), serotyping of E. coli isolates revealed, 26 (4.81%) strains O157: H7, 23 (4.26%) strains O111, 20 (3.70%) strains O113: H21, 10 (1.85%) strains O22: H8 and 7 (1.3%) strains O172: H21. Meanwhile, 17 (11.33%) strains of E. coli were recovered from raw meat samples, serotyping of E. coli isolates revealed, 6 (4%) strains O157: H7, 5 (3.33%) strains O111 and 4 (2.67%) strains O174: H2 and only two (1.33%) strains were identified as O22: H8. Shiga toxin2 was detected in 58 (67.44%) serotypes of E. coli recovered from milk samples and 16 (94.12%) serotypes of E. coli recovered from meat samples, while intimin gene was detected in 38 (44.186%) serotypes of E. coli recovered from milk samples and in 10 (58.82%) serotypes of E. coli recovered from meat samples. The results of this study revealed the efficiency of combination between serotyping and molecular typing of E. coli isolates recovered from food of animal origin for rapid detection and characterization of STEC.

Preparation of goat and rabbit anti-camel immunoglobulin G whole molecule labeled with horseradish peroxidase

Aim: As the labeled anti-camel immunoglobulins (Igs) with enzymes for enzyme-linked immunosorbent assay (ELISA) are unavailable in the Egyptian market, the present investigation was directed for developing local labeled anti-camel IgG with horseradish peroxidase (HRP) to save hard curacy. Materials and Methods: For purification of camel IgG whole molecule, camel sera was preliminary precipitated with 50% saturated ammonium sulfate and dialyzed against 15 mM phosphate-buffered saline pH 7.2 then concentrated. This preparation was further purified by protein A sepharose affinity column chromatography. The purity of the eluted camel IgG was tested by sodium dodecyl sulfate polyacrylamide gel electrophoresi. Anti-camel IgG was prepared by immunization of goats and rabbits separately, with purified camel IgG. The anti-camel IgG was purified by protein A sepharose affinity column chromatography. Whole molecule anti-camel IgG was conjugated with HRP using glutraldehyde based assay. Sensitivity and specificity of prepared conjugated secondary antibodies were detected using positive and negative camel serum samples reacted with different antigens in ELISA, respectively. The potency of prepared conjugated antibodies was evaluated compared with protein A HRP. The stability of the conjugate at −20°C during 1 year was assessed by ELISA. Results: The electrophoretic profile of camel IgG showed four bands of molecular weight 63, 52, 40 and 33 kDa. The recorded sensitivity and specificity of the product are 100%. Its potency is also 100% compared to 58-75% of commercial protein A HRP. The conjugates are stable for 1 year at −20°C as proved by ELISA. Conclusion: Collectively, this study introduces goat and rabbit anti-camel IgG whole molecules with simple, inexpensive method, with 100% sensitivity, 100% specificity and stability up to 1 year at −20°C. The important facet of the current study is saving hard curacy. Future investigations are necessary for preparation of IgG subclasses.

Polymerase chain reaction as an analytical tool in the diagnosis of cattle mastitis

E coli/Shigella exist as the major facultative anaerobe in the human gastrointestinal tract, but also can cause serious disease especially in young children, travelers and those that do not have access to clean water. E. coli/Shigella have been a model microbial system in use for decades, the advent of the genomic era has allowed unprecedented insight into the variation of this important species. Genome scale analysis has identified that approximately half of the E. coli/Shigella genome, ~2.2 Mbp, is highly conserved and represents an open genome structure that is constantly acquiring novel genetic material. Detailed studies have focused on the enterotoxigenic E. coli (ETEC) pathovar and the Shigella species. Genomic studies of each of these pathogens have revealed a complex and convoluted genomic evolution that is unique to each of the examples. Additionally, we have examined the ETEC transcriptional response to known virulence signals such as glucose concentration and intra-kingdom signals and have identified a number of candidate ETEC virulence factors. In addition to potential virulence factors, we have also identified novel genes that are conserved and may be involved in the regulation of virulence in this group of pathogens. Comparative genomics and transcriptomics has reached a new era where the number of strains that can be rapidly sequenced will begin to address epidemiological questions and become a more integrated component of the global health care system. David A. Rasko, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004N plant genes influence susceptibility to Agrobacterium-mediated transformation, but until recently no gene has been identified that globally regulates susceptibility. We identified MTF, an Arabidopsis myb transcription factor that negatively regulates transformation. MTF is down-regulated by cytokinins secreted by Agrobacterium, resulting in increased transformation susceptibility. In plants, cytokinins trigger a signaling cascade mediated by a two-component phosphorelay pathway consisting of the AHKs (Arabidopsis histidine kinases) and the ARRs (Arabidopsis response regulators). ahk3 and ahk4 mutants show attenuated transformation, indicating involvement of these primary cytokinin receptors in transformation. Of the several ARR mutants tested, only arr3 shows decreased transformation-susceptibility. One of the earliest transformation events is the attachment of bacteria to plant cells. In the hyper-transforming mtf mutant, one of the transcriptionally up-regulated genes is AT14a, which encodes an integrin domain-containing protein. AT14a is plasma membrane-localized and may mediate connections between the cell wall and the cytoskeleton. mtf mutants show increased bacterial attachment and transformation, whereas at14a mutants show lower Agrobacterium attachment and transformation. AT14a transgenic plants also show increased transformation and bacterial attachment. Thus, modulation of MTF expression via the cytokinin signaling pathway plays an important role in Agrobacteriummediated transformation via increased bacterial attachment. We identified putative MTF orthologs in the crop species rice, Brassica napus, B. rapa, B. oleracea, soybean, and maize. cDNAs of these putative orthologs functionally complement the Arabidopsis mtf1-4 mutant, whereas an unrelated myb transcription factor cDNA does not. Preliminary data indicate that RNAi directed against the rice MTF ortholog results in increased rice transformation susceptibility. Stanton B. Gelvin, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004A 18% of the global cancer burden has been attributed to infectious agents, with estimates ranging from 7% in developed countries to about 22% in developing countries. Chronic infections caused by the hepatitis B and C viruses, human papillomaviruses (HPV), and Helicobacter pylori (H. pylori) are reported to be responsible for approximately 15% of all human cancers. Interestingly, although many of the infectious agents that have been associated with cancer—such as HPV, EpsteinBarr virus (EBV), and H. pylori-are highly prevalent in the world, most infected individuals do not develop cancer but remain lifelong carriers. Malignancies associated with infectious agents may result from prolonged latency as a result of chronic infections. Pathogenic infections are necessary but are not sufficient for cancer initiation or progression. Cancer initiation may require additional cofactors, including secondary infections. Therefore, in patients with chronic infection with one agent, secondary coinfection with another agent may serve as an important co-factor that may cause cancer initiation and progression. Additionally, opportunistic co-infections could significantly inhibit response to cancer treatment and increase cancer mortality. Co‐infections are relatively common in areas with a high prevalence of infectious agents, especially in developing countries. These co-infections can cause an imbalance in the host immune system by affecting persistence of and susceptibility to malignant infections. Several articles have been published that focus on infectious agents and cancer. In this presentation, the role of infectious agents in malignancies, highlight the role of multiple/co-infections in cancer etiology, and review implications for cancer epidemiology will be discussed. Mukesh Verma, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004This study examined healthcare providers’ adherence to the national Tuberculosis guidelines (NTG) during the diagnosis and treatment of TB in Addis Ababa, Ethiopia using a descriptive, cross-sectional study design. Data were collected from 233 medical records using checklists. Adherence of healthcare providers to the NTG during the diagnosis of TB was 60.9% (n=67) for female and 56.1% (n=69) for male TB patients. However, 91.8% (n=101) female and 90.2% (n=111) male TB patients had been prescribed the correct numbers of anti-TB pills, complying with the NTG recommendations. There was an over-diagnosis of smear negative pulmonary Tuberculosis (PTB) as only 2.6% (n=2) of the 76 smear negative PTB patients were diagnosed correctly. Healthcare providers’ compliance with the NTG could be enhanced by providing appropriate in-service education, maintaining accurate records of all TB patients and providing supportive supervision to identify and address shortcomings.Background and aims Children with a lung abscess usually do well with antibiotics alone and surgical intervention is rarely needed. Standard practice is to use parenteral antibiotics until clinical symptoms abate and to follow with oral antibiotics for up to six weeks. The objective of this study was to observe and compare outcome, duration of antimicrobial treatment for lung abscess. Methods A prospective open, randomized clinical trial was conducted among 30 children aged 5 to 15 years with lung abscess and sequential antibiotic therapy either clindamycin (group 1; n=15) or ceftriaxone, flucloxacillin plus metronidazole (group 2; n =15) were administered until complete resolution of clinical and radiological abnormalities. Results Mean age was 11.5 years in group 1 and 11 years in group 2. Blood culture was negative in all cases but in sputum 33% cases staphylococcus aureus and 20% cases streptococcus pneumoniae was found and was sensitive to clindamycin, flucloxacillin and ceftriaxone. ESR exceeded 20 mm/hour in 94% and CRP exceeded 20 mg/L in 95% of the cases. ESR became normal in 21 days and CRP in 10 days and the cavity size on chest radiography was reduced after 14 days of treatment in first group but in second group CRP in 15 days, ESR in 28 days and reduced cavity size in 28 days. Mean duration of therapy was 21 days for first group and 39 days in second group. There were significant differences between the duration of treatment and outcome of the two groups (P<0.05. Conclusions Clindamycin appears to be effective short course treatment option in lung abscess.T global emergence of Clostridium difficile infection (CDI) has contributed to the recent surge in severe antibiotic-associated diarrhea and colonic inflammation. C. difficile produces two homologous glucosylating exotoxins, TcdA and TcdB, both of which are pathogenic and require neutralization to prevent disease occurrence. However, because of their large size and complex multifunctional domain structures, it has been a challenge to produce native recombinant toxins that may serve as vaccine candidates. Here, we describe a novel chimeric toxin vaccine that retains major neutralizing epitopes from both toxins and confers complete protection against primary and recurrent CDI in mice. Using a nonpathogenic Bacillus megaterium expression system, we generated glucosyltransferase-deficient holotoxins and demonstrated their loss of toxicity. The atoxic holotoxins induced potent antitoxin neutralizing antibodies showing little cross-immunogenicity or protection between TcdA and TcdB. To facilitate simultaneous protection against both toxins, we generated an active clostridial toxin chimera by switching the receptor binding domain of TcdB with that of TcdA. The toxin chimera was fully cytotoxic and showed potent proinflammatory activities. This toxicity was essentially abolished in a glucosyltransferase-deficient toxin chimera, cTxAB. Parenteral immunization of mice or hamsters with cTxAB induced rapid and potent neutralizing antibodies against both toxins. Complete and long-lasting disease protection was conferred by cTxAB vaccinations against both laboratory and hypervirulent C. difficile strains. Finally, prophylactic cTxAB vaccination prevented spore-induced disease relapse, which constitutes one of the most significant clinical issues in CDI. Thus, the rational design of recombinant chimeric toxins provides a novel approach for protecting individuals at high risk of developing CDI. Hanping Feng, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004T stable propagation of transcriptional states through numerous cell divisions is central to adaptability of all living organisms. This process, coined epigenetic inheritance, was first understood molecularly in lysogenic E. coli harboring a lambda prophage. Decades of research has revealed that all epigenetic pathways from bacteria to humans exploit the same molecular principles: specific and cooperative molecular interactions target specific regions of the chromosome, and create positive and negative feedback loops, respectively, which ultimately lock transcriptional states into a specific fate. The fission yeast heterochromatin has served as a faithful model for understanding the molecular mechanism of epigenetic inheritance in eukaryotes. Heterochromatin in S. pombe like other higher eukaryotes is demarcated by methylated lysine 9 of histone H3 (H3K9me). Several key silencing proteins, including heterochromatin protein 1 (HP1) bind to H3K9me and recruit repressive activities to heterochromatin. These repressive activities operate in cis by limiting RNA Pol II transcription and degradation of heterochromatic transcripts via the exosomeor RNAi-dependent mechanisms. In the fission yeast, a single histone methyltransferase, Clr4 belonging to the highly conserved Suv-39 family of proteins, methylates H3K9. Current models of epigenetic pathways posit that recruitment of Clr4 to heterochromatin is the initiating event for heterochromatin formation, tethering all repressive activities via H3K9me and downstream modifications. Here we show that several silencing proteins ‘prime’ chromatin for Clr4 recruitment, and are required for efficient H3K9 methylation. These data reveal a novel previously underappreciated step in heterochromatin formation, which precedes H3K9 methylation and regulates heterochromatin formation in eukaryotes. Mo Motamedi, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004T role of diet non-digestible bioactive compounds including phenolics and dietary fiber on the modulation of gut microbiota and the possible relationship with obesity is not fully understood. Our in vitro studies with colon fibroblast cells demonstrated that phenolic compounds decrease inflammation in colonic fibroblast cells in part mediated by induction of antioxidant enzymes that contributes to improve the immune response of colon fibroblast cells. In addition phenolics downregulated the TLR4 pathway, a known target for pathogens that promotes the expression of adhesion molecules that contributes to inflammatory bowel disease. In vitro and in vivo studies showed that fruit non-digestible bioactive compounds including dietary fiber and phenolic compounds induced a significant increase on the relative populations of health beneficial bacteria including Lactobacillus sp. and Bifidobacterium sp.; whereas the Enterobacteriaceae genera which include many of the more familiar pathogens were decreased. The link between phenolic compounds-gut microbiota-obesity was investigated using and in vivo animal model. Data from fecal DNA pyrosequencing analysis showed that Bacteroidetes, Ruminococcacea, and Lactobacilluswere in higher abundance in obese animals fed with fruit phenolics compared to phenolic absent controls. qRT-PCR data showed the phenolic-induced higher proportions of Faecalibacterium which has been reported to be in low levels in patients with Crohn’s Disease. These changes were accompanied by decreased weight gain; improved levels of metabolic syndrome and decreased levels of lipid peroxidation markers in plasma. Overall, these results suggest that protection against obesity and obesity related disorders is mediated by diet-induced altered composition of the distal intestinal microbiota. Giuliana D. Noratto, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004Abstract Objective The aim of this research was to study and evaluate the antimicrobialactivity of a novel 2-methylsulfanyl-[1,2,4]triazolo[1,5- a ]quinazoline and itsderivatives. Antibacterial activity of the target compounds was tested against avariety of species of Gram-positive bacteria such as Staphylococcus aureus ATCC29213, Bacillus subtilis ATCC6633, and Gram-negative bacteria such as Pseu-domonas aeruginosa ATCC27953 and Escherichia coli ATCC 25922. In additionsome yeast and fungi, Candida albicans NRRLY-477 and Aspergillus niger , respec-tively, were screened. Methods Antimicrobial tests were carried out by the agar well diffusion method,using 100 ml of suspension containing 1 ¥ 10 8 CFU/ml of pathological tested bac-teria, 1 ¥ 10 6 CFU/ml of yeast, and 1 ¥ 10 4 spore/ml of fungi spread on nutrientagar (NA), Sabourand dextrose agar (SDA), and potato dextrose agar (PDA),respectively. Key findings The minimum inhibitory concentration (MIC) of the tested com-pounds was determined using the broth double dilution method (serially dilutedtechnique) in proper nutrient. For comparison, ciprofloxacin and ketoconazolewere used as antibacterial and antifungal reference drugs, respectively. Com-poundsC continues to be a major public health threat, particularly in countries where safe drinking water, sanitation and hygiene are suboptimal. Toxigenic Vibrio cholerae, responsible for epidemic cholera, has two life styles, including (i) a transient and accidental occurrence in human intestine where it causes profuse diarrhea, and (ii) a ubiquitous occurrence in aquatic environments, including fresh, estuarine and marine waters. Although human acquire the disease by consumption of water contaminated with the bacterium, the genetic and physiologic basis of persistence of V. cholerae in the aquatic reservoirs, even during an ongoing cholera epidemic, is largely unknown. In response to stress such as in antibiotic stress, pathogenic bacteria are known to develop a “persister” phenotype which evades the stressful condition by stochastic mechanism. Using filter sterilized microcosm model, we recently demonstrated that, in response to nutrient stress,V. cholerae can select a subpopulation of bacterium, that, we termed as novel “persister” phenotype. The novel persister V. cholerae exhibited unprecedented bipolar and peritrichous flagella, diverse morphological cellular types, increased quorum sensing molecules, particularly CAI-1 molecule, and response to some nutrients while remained nonresponsive to others. We are currently examined other characteristics associated uniquely to persister V. cholerae and will be discussed in the proposed meeting. Afsar Ali, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004A Thesis Submitted to the School of Graduate Studies of the Addis Ababa University in Partial Fulfillment of the Requirements for the Degree of Master of Science in Medicinal Chemistry.B is amongst the top 5 causes of zoonotic disease worldwide. Infection is through ingestion, inhalation or contact exposure. Brucella is characterized as a class B pathogen by Centers of Disease Control and Prevention (CDC). Currently, there are no efficacious vaccines available in people. Available USDA approved vaccines for animals include strain B. abortus strain RB51 and B. melitensis Rev1. Protection is mediated by a strong innate and CD4 Th1, CD8 Tc1 immune response. If protective vaccines can be developed, disease in people and animals can be controlled. While strain RB51 protects in cattle, and against intraperitoneal challenge in mice, it does not protect against respiratory challenge. Therefore, we assessed the efficacy of strain RB51 combined with different TLR agonists, and O-side chain from LPS, to enhance protection against respiratory challenge with strain 2308. We hypothesized that TLR agonists and O-side chain would enhance protection. Strains RB51TLR2, RB51TLR4 and strain 19 provided significant protection in the lung. Protection from strain RB51+ TLR agonists was associated with increased IgG2a and IgG1 (BAL and serum), and increased IgA (serum). Splenocytes from strain RB51TLR2 vaccinated mice up-regulated antigen specific interferon-gamma and TNF-alpha production. Overall, this study demonstrates the ability of TLR agonists 2, and 4 to up-regulate strain RB51 mediated protection in the lung to respiratory challenge against strain 2308. Naveen Surendran, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004R studies have shown the efficacy of complex bacteriotherapy, such as fecal microbiota transplantation (FMT), for the treatment of antibiotic refractory Clostridium difficile colitis (C. diff). Serial FMTs have been indicated to potentially cure chronic inflammatory intestinal disorders, such as ulcerative colitis (UC) as well. In the meantime, this therapeutic option has been rarely explored in children while they suffer from treatment refractory C. diff and UC as well. We have recently developed pediatric protocols for intestinal microbiome transplantation (IMT, same as FMT) with frozen filtered stool preparations. The preparations were used with success in a case of moderate to severely active pediatric UC complicated by C. diff infection, and in a case of infliximab dependent UC where the patient was able to stop all conventional therapies. The therapeutic efforts were supported by 454 based pyrosequencing of the fecal and colonic mucosal microbiomes. Our findings will promote the establishment of complex (community based) bacteriotherapy in the management of pediatric gastrointestinal disorders. Richard Kellermayer, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004C inflammatory periodontal diseases (PD) are among the most common unresolved infections. Type 2 diabetes (T2D) is a similarly prevalent disease that represents a major risk factor for PD. Patients who develop T2D and PD suffer significant morbidity from the seemingly cyclical nature of one disease confounding the other. Mechanisms underlying the relationship between PD and T2D remain poorly defined, but one unifying link between T2D and PD is altered B lymphocyte function. Our new analyses using the standard P. gingivalis oral gavage model to induce PD in lean mice demonstrates B lymphocytes play minor roles in PD bone loss in this context. To the contrary, comparison of the obese/insulin resistant WT and B cell-null mouse responses to P. gingivalis gavage demonstrates B lymphocytes promote PD bone loss in obesity. Our data furthermore show that obesity-associated changes in B lymphocytes promote increased responses to oral pathogen ligands, as evidenced by elevated B cell production of TNF-α and MIP-2 when the cells are in vivo primed by a combination of obesity plus P. gingivalis, then in vitro challenged with P. gingivalis LPS or other toll-like receptor ligands. These data recapitulate our demonstration that B lymphocytes from PD patients hyper-produce TNF-α and the human MIP-2 ortholog (IL-8), thus suggest relevance of the mouse model to human PD. We conclude B lymphocytes promote obesity-associated PD and oral pathogen-associated systemic inflammation. Our data therefore raise the radical possibility that FDA-approved B cell depletion drugs may effectively break the feed-forward inflammatory loop between T2D and PD. Barbara Nikolajczyk, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004T formulation of antigens as particles in a size range mimicking infectious viruses and bacteria offers advantages over soluble antigens such as facilitated uptake by antigen-presenting cells (APCs), depot formation as well as co-delivery of antigens and immunomodulatory compounds to the same APC potentially controlling the type of immune response. A variety of bacteria are able to intracellularly produce polyester (polyhydroxybutyrate=PHB) inclusions which serve as carbon and energy reserve. Bioengineering of this natural polyester bead production process enabled the design and production of novel particulate vaccine delivery systems. A cost-effective, scalable and versatile particulate vaccine production process could be developed. This new technology offers an unprecedented design space accompanied with accelerated prototype development. Escherichia coli and the food-grade endotoxin-free bacterium Lactococcus lactis were engineered to produce spherical (PHB) inclusions which abundantly displayed the hepatitis C virus core (HCc) antigen or the two TB antigens, ESAT6 and Ag85A, respectively. In mice, the immune response induced by this antigen delivery system was compared to that induced by vaccination with only the soluble antigen. Vaccination site lesions were minimal in all mice vaccinated with PHB beads. Antigen displaying PHB beads stimulated an antigen-specific type 1 and 2 immune response. Moreover, a protective immunity was obtained in mice vaccinated against TB. Overall, this novel bead technology offers a safe and efficient vaccine delivery platform suitable for vaccination against viral infections and intracellular pathogens. Bernd H. A. Rehm, J Bacteriol Parasitol 2013, 4:4 http://dx.doi.org/10.4172/2155-9597.S1.004

Chlamydia species in free-living Cattle Egret (Bubulcus ibis) and Hoopoe (Upupa epops) in Egypt

Abstract Little information is available on the presence of chlamydia infection in wildlife. This study was conducted to assess the occurrence of chlamydiae in asymptomatic birds from two species of wild birds (Cattle Egret and Hoopoe) in Egypt. In the present study Chlamydiaceae was analyzed using Giemsa stain, Giménez stain, fluorescent antibody test (FAT) and PCR. The results of these techniques were compared with CFT for detecting Chlamydia psittaci antibodies among the examined birds. The results reveal that 96.4%, 81.8%, 89.1%, 80.0% and 58.2% of the examined samples were positive for chlamydiosis using PCR, Giemsa stain, Giménez stain, FA, and CFT respectively among Hoopoe. The percentages were 90.6%, 77.4%, 83.0%, 75.5% and 66.0% respectively for the previous tests among Cattle Egret birds. The results suggest that Cattle Egret and Hoopoe may be reservoir of Chlamydiaceae species and thus shed the organisms in their excreta. The shedding of chlamydiae by free living birds in Egypt may expose humans that come in contact with these birds to zoonotic risks.