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fecal DNA extraction

Asad Mohammadzadeh, Adel Spotin, Mahmoud Mahami-Oskouei, Ali Haghighi, Nozhat Zebardast, Kobra Kohansal
In the microevolutionary scales of Entamoeba isolates, the gene migration shows how Entamoeba spp. has epidemiologically drifted among border countries. Five hundred fecal samples were taken from patients suffering gastrointestinal disorders, abdominal pain, and diarrhea at Saggez, northwest Iran located within the border Iraq country. Following parasitological techniques, DNA samples were extracted and amplified by polymerase chain reaction (PCR) of 18S rRNA region to identify Entamoeba infections. To distinguish the Entamoeba spp...
October 20, 2016: Parasitology Research
Elizabeth K Mallott, Paul A Garber, Ripan S Malhi
OBJECTIVES: Invertebrate foraging strategies in nonhuman primates often require complex extractive foraging or prey detection techniques. As these skills take time to master, juveniles may have reduced foraging efficiency or concentrate their foraging efforts on easier to acquire prey than adults. MATERIALS AND METHODS: We use DNA barcoding, behavioral observations, and ecological data to assess age-based differences in invertebrate prey foraging strategies in a group of white-faced capuchins (Cebus capucinus) in northeastern Costa Rica...
October 5, 2016: American Journal of Physical Anthropology
Floriana Giorgio, Enzo Ierardi, Claudia Sorrentino, Mariabeatrice Principi, Michele Barone, Giuseppe Losurdo, Andrea Iannone, Antonio Giangaspero, Rosa Monno, Alfredo Di Leo
PURPOSE: Real-time polymerase chain reaction (RT-PCR) is a widely used technique for bacterial and viral infection diagnosis. Herein, we report our preliminary experience in retrieving H. pylori genetic sequences in stools and analyzing genotypic clarithromycin resistance by RT-PCR (noninvasive), with the aim of comparing this procedure with that performed on biopsy samples (invasive). MATERIALS AND METHODS: After 'in vitro' demonstration of H. pylori DNA detection from pure and stool-mixed bacteria, 52 consecutive patients at the first diagnosis of infection were investigated...
December 2016: Scandinavian Journal of Gastroenterology
Rosiléia Marinho de Quadros, Paulo Henrique Exterchoter Weiss, Sandra Marcia Tietz Marques, Luiz Claudio Miletti
Giardia duodenalis is an enteric parasite that has distinct genetic groups. Human infections are mainly caused by assemblages A and B, although sporadic infections by assemblages C and D have also been reported. Animals can be infected by a wide range of assemblages (A to H). The aim of this study is to identify the assemblages and sub-assemblages of G. duodenalis with zoonotic features in fecal samples of school-aged children, and in dogs that coexist in the same households in Lages, Santa Catarina, Brazil...
2016: Revista do Instituto de Medicina Tropical de São Paulo
Xinfeng Yan, Bo Feng, Peicheng Li, Zhaosheng Tang, Lin Wang
Background. Emerging evidences have shown a close interplay between obesity, diabetes, and intestinal flora disturbance. Dipeptidyl peptidase-4 inhibitor, exemplified by sitagliptin, is highly efficacious in treating type 2 diabetes (T2DM), yet little is known if sitagliptin exerts beneficial effects on microbiota associated with obesity and T2DM. We evaluated changes of gut microbiota following the induction of obesity and T2DM in a streptozotocin treated high fat/high carbohydrate fed (HF/HC-STZ) rat model and explored the effect of sitagliptin on gut microbiota for HF/HC-STZ rats...
2016: Journal of Diabetes Research
Kelley L Baumgartel, Maureen W Groer, Susan M Cohen, Dianxu Ren, Diane L Spatz, Yvette P Conley
BACKGROUND: Maternal interleukin (IL) single nucleotide polymorphisms (SNPs) are associated with obstetrical outcomes. Conversely, infant SNPs are associated with subsequent neonatal intensive care unit (NICU) outcomes. Little is known about relationships between maternal SNPs and neonatal outcomes. PURPOSE: To examine the relationships between maternal IL genotypes and neonatal outcomes. METHODS: An ancillary study was conducted among mothers (N = 63) who delivered very low-birth-weight infants (N = 74)...
September 6, 2016: Biological Research for Nursing
Lauren K Barron, Christopher P Gayer, Anne Roberts, Jamie M Golden, Bola G Aladegbami, Jun Guo, Christopher R Erwin, Brad W Warner
BACKGROUND: Intestinal failure-associated liver disease causes significant mortality in patients with short bowel syndrome. Steatosis, a major component of intestinal failure-associated liver disease has been shown to persist even after weaning from parenteral nutrition. We sought to determine whether steatosis occurs in our murine model of short bowel syndrome and whether steatosis was affected by manipulation of the intestinal microbiome. METHODS: Male C57BL6 mice underwent 50% small bowel resection and orogastric gavage with vancomycin or vehicle for 10 weeks...
August 31, 2016: Surgery
Maria Augusta Dario, Marina Silva Rodrigues, Juliana Helena da Silva Barros, Samanta Cristina das Chagas Xavier, Paulo Sérgio D'Andrea, André Luiz Rodrigues Roque, Ana Maria Jansen
BACKGROUND: Trypanosoma cruzi infection via oral route results in outbreaks or cases of acute Chagas disease (ACD) in different Brazilian regions and poses a novel epidemiological scenario. In the Espírito Santo state (southeastern Brazil), a fatal case of a patient with ACD led us to investigate the enzootic scenario to avoid the development of new cases. At the studied locality, Triatoma vitticeps exhibited high T. cruzi infection rates and frequently invaded residences. METHODS: Sylvatic and domestic mammals in the Rio da Prata locality, where the ACD case occurred, and in four surrounding areas (Baia Nova, Buenos Aires, Santa Rita and Todos os Santos) were examined and underwent parasitological and serological tests...
2016: Parasites & Vectors
Ana Raquel Madureira, Sara Nunes, Débora A Campos, João C Fernandes, Cláudia Marques, Monica Zuzarte, Beatriz Gullón, Luís M Rodríguez-Alcalá, Conceição Calhau, Bruno Sarmento, Ana Maria Gomes, Maria Manuela Pintado, Flávio Reis
Rosmarinic acid (RA) possesses several protective bioactivities that have attracted increasing interest by nutraceutical/pharmaceutical industries. Considering the reduced bioavailability after oral use, effective (and safe) delivery systems are crucial to protect RA from gastrointestinal degradation. This study aims to characterize the safety profile of solid lipid nanoparticles produced with Witepsol and Carnauba waxes and loaded with RA, using in vitro and in vivo approaches, focused on genotoxicity and cytotoxicity assays, redox status markers, hematological and biochemical profile, liver and kidney function, gut bacterial microbiota, and fecal fatty acids composition...
2016: International Journal of Nanomedicine
Ayse Seyer, Djursun Karasartova, Emrah Ruh, Ayse Semra Güreser, Turgut Imir, Aysegul Taylan-Ozkan
PCR and DNA sequencing are currently the diagnostic methods of choice for detection of Blastocystis spp. and their suptypes. Fresh or frozen stool samples have disadvantages in terms of several aspects such as transportation, storage, and existence of PCR inhibitors. Filter paper technology may provide a solution to these issues. The aim of the present study was to detect Blastocystis spp. and their subtypes by employing two different preservation methods: conventional frozen stool (FS) and dried stool spots on filter paper (DSSFP)...
August 17, 2016: Parasitology Research
Sandra Janezic
PCR-ribotyping, a method based on heterogeneity of ribosomal intergenic spacer region, is the preferred method for genotyping of Clostridium difficile. Standardly used procedure for PCR-ribotyping is culturing of C. difficile from fecal samples and subsequent typing. In this chapter, we describe a modified PCR-ribotyping method for direct detection of PCR-ribotypes directly in total stool DNA extract, without prior need to isolate C. difficile.
2016: Methods in Molecular Biology
Amandine Laffite, Pitchouna I Kilunga, John M Kayembe, Naresh Devarajan, Crispin K Mulaji, Gregory Giuliani, Vera I Slaveykova, John Poté
Data concerning the occurrence of emerging biological contaminants such as antibiotic resistance genes (ARGs) and fecal indicator bacteria (FIB) in aquatic environments in Sub-Saharan African countries is limited. On the other hand, antibiotic resistance remains a worldwide problem which may pose serious potential risks to human and animal health. Consequently, there is a growing number of reports concerning the prevalence and dissemination of these contaminants into various environmental compartments. Sediments provide the opportunity to reconstruct the pollution history and evaluate impacts so this study investigates the abundance and distribution of toxic metals, FIB, and ARGs released from hospital effluent wastewaters and their presence in river sediments receiving systems...
2016: Frontiers in Microbiology
Marie-Eve Fecteau, Dipti W Pitta, Bonnie Vecchiarelli, Nagaraju Indugu, Sanjay Kumar, Susan C Gallagher, Terry L Fyock, Raymond W Sweeney
Johne's disease (JD) is a chronic, intestinal infection of cattle, caused by Mycobacterium avium subsp. paratuberculosis (MAP). It results in granulomatous inflammation of the intestinal lining, leading to malabsorption, diarrhea, and weight loss. Crohn's disease (CD), a chronic, inflammatory gastrointestinal disease of humans, has many clinical and pathologic similarities to JD. Dysbiosis of the enteric microbiota has been demonstrated in CD patients. It is speculated that this dysbiosis may contribute to the intestinal inflammation observed in those patients...
2016: PloS One
Beatriz Garcia-Morante, Judit J Pénzes, Taiana Costa, Jaime Martorell, Jorge Martínez
A 2-year-old female, spur-thighed tortoise (Testudo graeca) was presented with poor body condition (1/5) and weakness. Fecal analysis revealed large numbers of oxyurid-like eggs, and radiographs were compatible with gastrointestinal obstruction. Despite supportive medical treatment, the animal died. At gross examination, an intestinal obstruction was confirmed. Histopathology revealed severe hyperplastic esophagitis and stomatitis with marked epithelial cytomegaly and enormous basophilic intranuclear inclusion bodies...
September 2016: Journal of Veterinary Diagnostic Investigation
Tatsuya Ide, Natsumi Kanzaki, Wakako Ohmura, Kimiko Okabe
A rapid and technically simple molecular detection method was developed for the western drywood termite, Incisitermes minor (Hagen), which is among the most harmful pests of wooden architectures. The method features DNA extraction from fecal pellets and species-specific loop-mediated isothermal amplification (LAMP) of DNA. As drywood termites have low moisture requirements, they easily infest artificial wooden products, and are likely to spread by human transportation of infested wood and wooden products. Rapid detection and accurate identification are essential tools for termite eradication at new sites of introduction...
July 29, 2016: Journal of Economic Entomology
Ali H D Janabi, Lee J Kerkhof, Lora R McGuinness, Amy S Biddle, Kenneth H McKeever
There are many choices for methods of extracting bacterial DNA for Next Generation Sequencing (NGS) from fecal samples. Here, we compare our modifications of a phenol/chloroform extraction method plus an inhibitor removal solution (C3) (ph/Chl+C3) to the PowerFecal® DNA Isolation Kit (MoBio-K). DNA quality and quantity coupled to NGS results were used to assess differences in relative abundance, Shannon diversity index, unique species, and principle coordinate analysis (PCoA) between biological replicates...
October 2016: Journal of Microbiological Methods
Nozhat Zebardast, Farshid Yeganeh, Mohammad Javad Gharavi, Alireza Abadi, Seyyed Javad Seyyed Tabaei, Ali Haghighi
Entamoeba histolytica, Giardia lamblia and Cryptosporidium spp. are common causes of diarrheal and intestinal diseases all over the world. Microscopic methods are useful in the diagnosis of intestinal parasites (IPs), but their sensitivity was assessed approximately 60 percent. Recently, molecular techniques have been used increasingly for the identification and characterization of the parasites. Among those, in this study we have used multiplex PCR and Real-time PCR with melting curve analysis (qPCR-MCA) for simultaneous detection and differentiation of E...
October 2016: Acta Tropica
Kilaza Samson Mwaikono, Solomon Maina, Aswathy Sebastian, Megan Schilling, Vivek Kapur, Paul Gwakisa
BACKGROUND: Multiple types of solid waste in developing countries is disposed of together in dumpsites where there is interaction between humans, animals and the bacteria in the waste. To study the bacteria at the dumpsite and the associated risks, previous studies have focused on culturable, leaving behind a great number of unculturable bacteria. This study focuses on a more comprehensive approach to study bacteria at the dumpsite. Since the site comprised of unsorted wastes, a qualitative survey was first performed to identify the variety of solid waste as this has influence on the microbial composition...
2016: BMC Microbiology
Miguel Saavedra, Inés Zulantay, Werner Apt, Juan Castillo, Eduardo Araya, Gabriela Martínez, Jorge Rodríguez
BACKGROUND: Trypanosoma cruzi multiplies and differentiates in the digestive tract of triatomine insects. Xenodiagnosis (XD) is a parasitological tool in which the insect vectors acts as a biological culture medium to amplify and detect T. cruzi infection in mammals. The sensitivity of XD has been overcome by the application of PCR in fecal samples (FS) of XD (PCR-XD). In this study, T. cruzi amplified in Triatoma infestans fed by XD on individuals with chronic Chagas disease (CChD) is quantified by real-time PCR (qPCR-XD)...
2016: Parasites & Vectors
Premalatha Pushpanathan, Padma Srikanth, Krishna G Seshadri, Sribal Selvarajan, Ravi Shankar Pitani, Thomas David Kumar, R Janarthanan
BACKGROUND: Type 2 diabetes mellitus (T2DM) and obesity are associated with changes in gut microbiota and characterized by chronic low-grade inflammation. Monocyte chemoattractant protein-1 (MCP-1) and interferon gamma (IFNγ) are proinflammatory cytokines which play an important role in the development of T2DM. We undertook this study to analyze the gut microbiota of T2DM and nondiabetic subjects and to determine the profile of MCP 1 and IFNγ in the same subjects attending a tertiary care center in Chennai, Tamil Nadu, India...
July 2016: Indian Journal of Endocrinology and Metabolism
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