Optimization of a fed-batch fermentation process for production of bleomycin by Streptomyces mobaraensis ATCC 15003

The objective of this work was to optimize inoculum size and pH for rapid production of bleomycin by Streptomyces mobaraensis ATCC 15003 by fed-batch fermentation. In both batch- and fed-batch fermentations, neither production period for bleomycin by S. mobaraensis nor the amount of BLM were affected by increasing the inoculum size from regular 10 to 30% (v/v) level. A fed-batch bioreactor not only shortened the lag phase of BLM production from 114 to 60 h, but also fed-batch fermentationenhanced BLM production when combined with an appropriate pH profile with no effect on the amount produced. Due to the substrate inhibition that takes place at high levels of carbon source, fed-batch fermentation was proposed as a better alternative for BLM production. The combined effects of batch and fed-batch fermentation and various pH profiles on BLM production in a bioreactor were evaluated. The tested pH profiles included; (1) a constant pH profile at 6.8 (profile1); (2) a constant pH profile witha period of auto-acidification for 72 h (profile2) and (3) a step-wise pH profile with pH adjustment every 24 h (profile3). When profile 3 was applied, fed-batch fermentation enhanced BLM production in the bioreactor and yielded about two-fold higher BLM concentration than the irrespective batch fermentation. On the other hand, constant pH profile or that which included one period of autoacidification (profiles 1 and 2) resulted in an insignificantly difference BLM production in fed-batch fermentation. Overall, this study suggested that fed-batch fermentation can be successfully used to enhance BLM production in bioreactor especially with fluctuated pH-profile.Key words: Bleomycin, Streptomyces mobaraensis, rapid production, fed-batch fermentation

Using molecular techniques for rapid detection of Salmonella serovars in frozen chicken and chicken products collected from Riyadh, Saudi Arabia

The current study was aimed to investigate the incidence of different Salmonella serovars in chicken products either from local or imported source. A total of 152 samples of chicken and chicken productswere collected from different retail establishment markets in Riyadh, KSA including 38 local whole frozen chickens, 62 imported whole frozen chickens, 22 whole poultry eggs and 30 local chicken cuts samples and examined by standard microbiological techniques (SMT). Salmonella isolation revealed a total percentage of 5.92%; chicken cuts revealed a high incidence among the examined samples (10%), followed by local frozen chickens and imported frozen chicken samples with incidence of 7.89 and 4.83%, respectively. For this experiment, the whole chicken eggs were negative for Salmonella species by SMT. Salmonella enteritidis was dominating among the recovered Salmonella serovars, followed bySalmonella typhimurium, while only two strains of Salmonella agona and Salmonella newport were isolated. The PCR assay combined with Rappaport- Vassiliadis (RV) selective broth (PCR-RV) for the detection of Salmonella species in the collected field samples revealed the same positive samples directly from the imported frozen chickens and whole chicken eggs which gave negative results by SMT. Thus PCR-RV technique is rapid, time saving and applicable to detect Salmonella serovarsdirectly from chicken samples

Multiplex polymerase chain reaction for detection and characterization of shiga toxigenic Escherichia coli (STEC)

Escherichia coli is ubiquitous in the cow's environment that is contaminated by feces, and it is also a frequent cause of bovine mastitis. Thus, the present study was targeted at the rapid detection and characterization of shiga toxigenic E. coli (STEC) in bovine fecal and milk samples. Twenty two strains of E. coli (39.29%) were isolated from 56 diarrheic calves, while only 5 strains (20.83%) were isolated from apparently normal contact calves. Moreover, 20 strains of E. coli (25%) were isolated from milk samples collected from 80 animals suffering from mastitis and subclinical mastitis. E. coli serovars yielded from bacteriological examination of milk samples were similar to that of fecal samples. Serogroup-specific multiplex polymerase chain reaction (PCR) assay could detect all the bacteriologically positive samples as well as 4 strains (7.98%), O157:H7 and 3 strains (5.36%), O111 from diarrheic calves and 2 strains (8.33%), O111 from normal calves. Such samples were proved to be negative by bacteriological examination. Multiplex PCR for detection of genes encoding accessorySTEC virulence factors, such as shiga toxin type-2 (stx2) and intimin gene (eaeA) revealed the specificity of such gene to O157:H7 serovars and small number of other sero-groups

Evaluation of the protective efficacy of immunoglobulin Y (IgY- antibodies) prepared against Walterinnesia aegyptia snake venom in Saudi Arabia

Four groups of eight chickens were immunized intramuscularly with Walterinnesia aegyptia snake venoms mixed with Freund's complete adjuvant during the period from 1st October 2009 to 1st October 2011 at the Center of Excellence in Biotechnology Research, King Saud University, Saudi Arabia. Three weeks later, the injections were repeated with the venoms in incomplete Freund's adjuvant. Three boosters were given with the venoms at three weeks intervals. The immunoglobulin Y (IgY)-antibodies was extracted by ammonium sulphate-caprylic acid method, the antibody titer were tested by enzyme linked immunosorbant assay and the protective efficacies of the extracted immunoglobulins were performed. IgY-preparation extracted by ammonium sulphate-caprylic acid method showed lack of low molecular weight bands (non-immunoglobulin proteins) and the bands representing IgY-antibodies, which have molecular weights ranging from 180 to 200 kDa, appeared sharp and clear. Moreover, evaluation of the protective value of the IgY - antibodies prepared revealed that, one milliliter of extracted IgY-antibodies containing 15 mg/ml anti-W. aegyptia venom specific IgY could produce 100% protection against 50 LD50 and 75% protection against 60 LD50. Laying hens could be used as an alternative source of polyclonal antibodies against W. aegyptia snake venoms due to several advantages as compared with mammals traditionally used for such purpose.Keywords: Snake venom, Walterinnesia aegyptia, immunoglobulins Y, protective efficacy, caprylic aci

The epidemiology of dengue outbreaks in 2016 and 2017 in Ouagadougou, Burkina Faso.

BACKGROUND: Dengue is prevalent in as many as 128 countries with more than 100 million clinical episodes reported annually and four billion people estimated to be at risk. While dengue fever is systematically diagnosed in large parts of Asia and South America, the disease burden in Africa is less well investigated. This report describes two consecutive dengue outbreaks in Ouagadougou, Burkina Faso in 2016 and 2017. METHODS: Blood samples of febrile patients received at Schiphra laboratory in Ouagadougou, Burkina Faso, were screened for dengue infection using SD Bioline Dengue Duo rapid diagnostic test kits (Standard Diagnostics, Suwon, Republic of Korea). RESULTS: A total of 1,397 and 1,882 cases were reported by a single laboratory in 2016 and 2017, respectively. Most cases were at least 15 years of age and the results corroborated reports from WHO indicating the circulation of three dengue virus serotypes in Burkina Faso. CONCLUSION: This study complements data from other, simultaneously conducted surveillance efforts, and indicates that the dengue disease burden might be underestimated in sub-Saharan African nations. Dengue surveillance should be enhanced in African settings to determine the burden more accurately, and accelerated efforts towards a dengue vaccine should be put in place

Barking up the same tree: a comparison of ethnomedicine and canine ethnoveterinary medicine among the Aguaruna

<p>Abstract</p> <p>Background</p> <p>This work focuses on plant-based preparations that the Aguaruna Jivaro of Peru give to hunting dogs. Many plants are considered to improve dogs' sense of smell or stimulate them to hunt better, while others treat common illnesses that prevent dogs from hunting. This work places canine ethnoveterinary medicine within the larger context of Aguaruna ethnomedicine, by testing the following hypotheses: H1 -- Plants that the Aguaruna use to treat dogs will be the same plants that they use to treat people and H2 -- Plants that are used to treat both people and dogs will be used for the same illnesses in both cases.</p> <p>Methods</p> <p>Structured interviews with nine key informants were carried out in 2007, in Aguaruna communities in the Peruvian department of Amazonas. Informants provided freelists of plants given to dogs and explained the purpose, preparation and route of administration used. For each plant, informants also described any uses for treating people. Botanical voucher specimens were collected and additional informal observations were made, accompanying people on hunting trips.</p> <p>Results</p> <p>Out of 35 plant species given to dogs, 29 (83%) are also given to humans for some medicinal purpose, while five are used only for dogs. However, the same plant is used to treat the same illness in both humans and dogs in only 53% of the cases. Forty-three percent of plants used to treat a particular illness for both dogs and people are administered in the same manner for both.</p> <p>Conclusion</p> <p>Results suggest that Aguaruna canine ethnoveterinary medicine is, at least partly, an independent cognitive domain. Some of the difference in plant use between dogs and people can be explained by the fact that certain diseases mentioned only apply to dogs. Although reports of canine ethnoveterinary medicine are very sparse in the literature, Aguaruna practices show some similarities with a few trends reported for other Amazonian societies, particularly, in the prevalence of the nasal route of administration, the use of plant-based psychoactives and in the importance of ants and wasps, in some form, for training dogs.</p

Association of Panton Valentine Leukocidin (PVL) genes with methicillin resistant Staphylococcus aureus (MRSA) in Western Nepal: a matter of concern for community infections (a hospital based prospective study)

BACKGROUND: Methicillin resistant Staphylococcus aureus (MRSA) is a major human pathogen associated with nosocomial and community infections. Panton Valentine leukocidin (PVL) is considered one of the important virulence factors of S. aureus responsible for destruction of white blood cells, necrosis and apoptosis and as a marker of community acquired MRSA. This study was aimed to determine the prevalence of PVL genes among MRSA isolates and to check the reliability of PVL as marker of community acquired MRSA isolates from Western Nepal. METHODS: A total of 400 strains of S. aureus were collected from clinical specimens and various units (Operation Theater, Intensive Care Units) of the hospital and 139 of these had been confirmed as MRSA by previous study. Multiplex PCR was used to detect mecA and PVL genes. Clinical data as well as antimicrobial susceptibility data was analyzed and compared among PVL positive and negative MRSA isolates. RESULTS: Out of 139 MRSA isolates, 79 (56.8 %) were PVL positive. The majority of the community acquired MRSA (90.4 %) were PVL positive (Positive predictive value: 94.9 % and negative predictive value: 86.6 %), while PVL was detected only in 4 (7.1 %) hospital associated MRSA strains. None of the MRSA isolates from hospital environment was found positive for the PVL genes. The majority of the PVL positive strains (75.5 %) were isolated from pus samples. Antibiotic resistance among PVL negative MRSA isolates was found higher as compared to PVL positive MRSA. CONCLUSION: Our study showed high prevalence of PVL among community acquired MRSA isolates. Absence of PVL among MRSA isolates from hospital environment indicates its poor association with hospital acquired MRSA and therefore, PVL may be used a marker for community acquired MRSA. This is first study from Nepal, to test PVL among MRSA isolates from hospital environment. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12879-016-1531-1) contains supplementary material, which is available to authorized users

Measurement of the Bottom-Strange Meson Mixing Phase in the Full CDF Data Set

We report a measurement of the bottom-strange meson mixing phase \beta_s using the time evolution of B0_s -> J/\psi (->\mu+\mu-) \phi (-> K+ K-) decays in which the quark-flavor content of the bottom-strange meson is identified at production. This measurement uses the full data set of proton-antiproton collisions at sqrt(s)= 1.96 TeV collected by the Collider Detector experiment at the Fermilab Tevatron, corresponding to 9.6 fb-1 of integrated luminosity. We report confidence regions in the two-dimensional space of \beta_s and the B0_s decay-width difference \Delta\Gamma_s, and measure \beta_s in [-\pi/2, -1.51] U [-0.06, 0.30] U [1.26, \pi/2] at the 68% confidence level, in agreement with the standard model expectation. Assuming the standard model value of \beta_s, we also determine \Delta\Gamma_s = 0.068 +- 0.026 (stat) +- 0.009 (syst) ps-1 and the mean B0_s lifetime, \tau_s = 1.528 +- 0.019 (stat) +- 0.009 (syst) ps, which are consistent and competitive with determinations by other experiments.Comment: 8 pages, 2 figures, Phys. Rev. Lett 109, 171802 (2012

Controversy and consensus on indications for sperm DNA fragmentation testing in male infertility: a global survey, current guidelines, and expert recommendations.

PURPOSE: Sperm DNA fragmentation (SDF) testing was recently added to the sixth edition of the World Health Organization laboratory manual for the examination and processing of human semen. Many conditions and risk factors have been associated with elevated SDF; therefore, it is important to identify the population of infertile men who might benefit from this test. The purpose of this study was to investigate global practices related to indications for SDF testing, compare the relevant professional society guideline recommendations, and provide expert recommendations. MATERIALS AND METHODS: Clinicians managing male infertility were invited to take part in a global online survey on SDF clinical practices. This was conducted following the CHERRIES checklist criteria. The responses were compared to professional society guideline recommendations related to SDF and the appropriate available evidence. Expert recommendations on indications for SDF testing were then formulated, and the Delphi method was used to reach consensus. RESULTS: The survey was completed by 436 experts from 55 countries. Almost 75% of respondents test for SDF in all or some men with unexplained or idiopathic infertility, 39% order it routinely in the work-up of recurrent pregnancy loss (RPL), and 62.2% investigate SDF in smokers. While 47% of reproductive urologists test SDF to support the decision for varicocele repair surgery when conventional semen parameters are normal, significantly fewer general urologists (23%; p=0.008) do the same. Nearly 70% would assess SDF before assisted reproductive technologies (ART), either always or for certain conditions. Recurrent ART failure is a common indication for SDF testing. Very few society recommendations were found regarding SDF testing. CONCLUSIONS: This article presents the largest global survey on the indications for SDF testing in infertile men, and demonstrates diverse practices. Furthermore, it highlights the paucity of professional society guideline recommendations. Expert recommendations are proposed to help guide clinicians

Controversy and consensus on the management of elevated sperm DNA fragmentation in male infertility: a global survey, current guidelines, and expert recommendations

PURPOSE: Sperm DNA fragmentation (SDF) has been associated with male infertility and poor outcomes of assisted reproductive technology (ART). The purpose of this study was to investigate global practices related to the management of elevated SDF in infertile men, summarize the relevant professional society recommendations, and provide expert recommendations for managing this condition. MATERIALS AND METHODS: An online global survey on clinical practices related to SDF was disseminated to reproductive clinicians, according to the CHERRIES checklist criteria. Management protocols for various conditions associated with SDF were captured and compared to the relevant recommendations in professional society guidelines and the appropriate available evidence. Expert recommendations and consensus on the management of infertile men with elevated SDF were then formulated and adapted using the Delphi method. RESULTS: A total of 436 experts from 55 different countries submitted responses. As an initial approach, 79.1% of reproductive experts recommend lifestyle modifications for infertile men with elevated SDF, and 76.9% prescribe empiric antioxidants. Regarding antioxidant duration, 39.3% recommend 4-6 months and 38.1% recommend 3 months. For men with unexplained or idiopathic infertility, and couples experiencing recurrent miscarriages associated with elevated SDF, most respondents refer to ART 6 months after failure of conservative and empiric medical management. Infertile men with clinical varicocele, normal conventional semen parameters, and elevated SDF are offered varicocele repair immediately after diagnosis by 31.4%, and after failure of antioxidants and conservative measures by 40.9%. Sperm selection techniques and testicular sperm extraction are also management options for couples undergoing ART. For most questions, heterogenous practices were demonstrated. CONCLUSIONS: This paper presents the results of a large global survey on the management of infertile men with elevated SDF and reveals a lack of consensus among clinicians. Furthermore, it demonstrates the scarcity of professional society guidelines in this regard and attempts to highlight the relevant evidence. Expert recommendations are proposed to help guide clinicians