Loop-Mediated isothermal amplification in human Cytomegalovirus diagnostic

The pathogenicity and the consequences of human cytomegalovirus (HCMV) infection in immunocompromised persons are reported in the literature and, HCM Viral DNA detection is the reliable indicator of infection especially to study its reactivation after organ transplantation in the recipient patients. Congenital infections in pregnant women cause opportunistic diseases of the central nervous system in new-born.  HCMV increases suffering and death in patients with chronic diseases like diabetes, tuberculosis, hepatitis C or patients with immunodeficient HIV/AIDS. Diagnostic methods are present in medical analysis laboratories either by immunofluorescence, Chemiluminescence, immunochromatography, western blot or by molecular biology techniques such as PCR are used to detect qualitatively and/or quantitatively IgG and IgM antibodies in the organism using serum or plasma as samples. The follow-up and treatment of patients are carried out in case the clinical data correlate with specific pathologies and symptoms, thus, the analysis of cytomegalovirus by PCR is requested to be sure of the cause of this pathology and to eliminate other doubts because this virus shares genetic and pathological similarity with Epstein-Barr virus (EBV), Varicella-Zoster virus (VZV) and Herpes Simplex Virus (HSV) as they are all of the same family of the Herpesviridae. The viral load quantified by PCR is a key element for monitoring and treatment. Molecular biology researchers have developed different PCR amplification techniques to answer the question of sensitivity, efficiency, and speed of this technique, and more recently in 2000, Notomi and the EIKEN Chemical Co. Ltd. developed a method of amplification under isothermal conditions named LAMP (Loop-mediated isothermal amplification) which combines speed and easy access for all laboratories that want to use only the less expensive equipment like those used with real-time PCR, later this technique was validated by the World Health Organization (WHO) and the organization recommended it to be used as an alternative in research and diagnostics. In this bibliographic study, we will exploit the contribution of the LAMP method in the diagnosis of human cytomegalovirus

Marine biomolecules: a promising approach in therapy and biotechnology

The marine environment is characterized by a wide diversity of microorganisms among which marine bacteria. To insure their survival in hostile conditions where they face high competition with pathogenic microorganisms, they produce various kinds of bioactive molecules within biofilms with unique structural and functional features. As example: marine peptides which provide a broad spectrum of antimicrobial, antitumoral, antiviral and anti-inflammatory activities, in addition to marine exopolysaccharides showing antifouling and antifungal activities, immunomodulatory properties, emulsion stabilization capacity with other various potentials. Some biofilms have shown a beneficial role for aquaculture, among which enhancement of growth performance and improvement of water quality, while others are threatening not only aquaculture and maritime fields, but also medicine and food industry. Thus, marine bioactive compounds are promising preventing agents for the establishment and growth of fouling microorganisms, which may be useful in different fields in order to decrease economic losses and avoid foodborne illnesses. DOI: http://dx.doi.org/10.5281/zenodo.438415

Marine biomolecules: a promising approach in therapy and biotechnology

The marine environment is characterized by a wide diversity of microorganisms among which marine bacteria. To insure their survival in hostile conditions where they face high competition with pathogenic microorganisms, they produce various kinds of bioactive molecules within biofilms with unique structural and functional features. As example: marine peptides which provide a broad spectrum of antimicrobial, antitumoral, antiviral and anti-inflammatory activities, in addition to marine exopolysaccharides showing antifouling and antifungal activities, immunomodulatory properties, emulsion stabilization capacity with other various potentials. Some biofilms have shown a beneficial role for aquaculture, among which enhancement of growth performance and improvement of water quality, while others are threatening not only aquaculture and maritime fields, but also medicine and food industry. Thus, marine bioactive compounds are promising preventing agents for the establishment and growth of fouling microorganisms, which may be useful in different fields in order to decrease economic losses and avoid foodborne illnesses. DOI: http://dx.doi.org/10.5281/zenodo.438415

Comparability of CMV DNA Extraction Methods and Validation of Viral Load

Human cytomegalovirus is a herpesvirus that has a worldwide seroprevalence of more than 60% of adults in developed countries and 90% in developing countries. Severe disabilities in newborns are characteristic of the human cytomegalovirus congenital infection, and this virus is implicated in graft rejection in transplant patients. To treat and follow-up the infection, the CMVPCR viral loads are required, and the DNA extraction step remains very important; however, the quantity, quality, and purity of extracted DNA from different biological fluids influence the results of PCR amplification, that is why for reliable results, the choice of nucleic acid extraction methods requires careful attention. Materials and methods: In this study, we compare 4 protocols, I (EZ1 DSP Virus kit), II (EZ1 Virus mini kit), III (QIAamp DSP virus kit), and IV (heating); the extractions are made from plasma collected on EDTA tubes, and the concentration of extracted DNA was measured on NanoDrop Lite followed by real-time CMVPCR using an Artus CMV QS-RGQ kit. All protocols are performed following the manufacturer’s instructions. Results: This study is conducted on the samples of 135 transplant patients whose follow-up medical tests related to human cytomegalovirus infection; since most of the CMVPCR results are negative, we have chosen the 10 CMVPCR positive samples and 2 negative samples as controls to conduct this comparison study. By using NanoDrop Lite to evaluate the DNA concentration, the yield of extracted DNA is higher in our heating protocol than other protocols, the EZ1 DSP virus kit and EZ1 Virus mini kit show homogeneous quantities, and the QIAamp DSP virus kit shows very low DNA yields. Comparing cycle threshold and viral loads by real-time PCR, all these protocols identified negative samples (100%), and the previously positive samples used were as follows: protocol IV (90%), protocol II (60%), and protocol I (40%). QIAamp DSP virus kit results were not real-time PCR applicable and were non-conclusive because of the low DNA yields. Conclusion: Our developed heating method (protocol IV) is very effective, reliable, simple, fast, and cheap compared to the other protocols in our study

Study of the Biochemical Biodiversity of Camel Milk

Due to its balanced composition in basic nutrients, its richness in vitamins and in minerals, camel milk has a special place in the daily diet of southern populations. The present study aimed at examining some physicochemical and biochemical characteristics of camel milk (Camelus dromedarius) in Morocco. To achieve this aim, three batches of samples of camel milk were collected from various regions (Dakhla, Errachida, and Fès-Meknes) undergoing physicochemical and biochemical analyses. Results showed that Moroccan camel milk is characterized by slight hydronium potential (pH=6.5), low Dornic acidity (15°D), low density (1.029 kg/l), and high content of ashes (8.06 g/l). Likewise, samples had a high content of macronutrients (Fats: 34.09 g/l; Proteins: 32.4 g/l; Sugar: 49.8 g/l) and micronutrients (Vitamin C: 27.53 mg/l; Flavonoids: 29.05 mg EQ/l: total phenolic compounds: 35.45mg GAE/l). In this respect, working on multiple specimens from different Moroccan regions highlighted an analytical diversity from the south to the north. Comparative study of samples from numerous territories all over the world has confirmed this diversity. North African milk is characterized by high content of proteins, fats, and sugar. On the other hand, oriental milk is peculiarized by high vitamin C content

The Life Hidden Inside Caves: Ecological and Economic Importance of Bat Guano

Bats are emblematic hosts of caves. These small flying mammals deserve special attention because their presence has a great economic and ecological impact; they introduce organic matter, the guano, in the ecosystem they live in. Indeed, “guano” (a Quechua word meaning “fertilizer”) is the accumulation of their fecal matter (excreta); its deposition can reach several meters. The composition of guano is influenced by the bat’s food. In addition to its role within the caves, the bat guano is exploited for various purposes; it is used as an effective fertilizer for the cultivation of plants because of having exceptionally high content of nitrogen, phosphate, and potassium and also for the improvement of detergents and other products of great value for humans. The bat guano hosts various classes of microorganisms (viruses, bacteria, algae, fungi, and protists), which are adapted to the cave environment. Since guano is highly acidic, these microorganisms can be considered as extremophiles. They produce functional organic compounds in extreme conditions that could be of interest not only in the drug industry but also in different biotechnological areas. Here, we review already available information on the ecological and economic effects of bats and their guano. We report their food preferences, foraging behaviors, and environmental impacts. Information on these aspects may be useful in finding a solution about protection and preservation of bat populations

Effect of Origanum vulgare Essential Oil Supplementation on the Advanced Parameters of Mobility and on the Integrity of Human Sperm DNA

The reduced sperm mobility is one of the most important causes of male infertility. Several reports have indicated that the treatment of subnormal sperm samples with certain agents prior to artificial insemination significantly improves the fertilizing potential of sperm. We have among others some stimulants such as pentoxifylline, relaxin, prostaglandin E, and diltiazem. In our precedent work, we had tested the effect of supplementation with three essential oils, namely, sage (Salvia officinalis), oregano (Origanum vulgare), and eucalyptus (Eucalyptus globulus), on sperm cell mobility and vitality. Oregano oil had shown interesting biological properties by giving the best values of progressive mobility and vitality. In this study, we aim to verify the effect of oregano oil supplementation on the advanced parameters of mobility and on the integrity of the sperm DNA of 25 male infertile volunteers. Our results showed that oregano oil over an incubation period of 5 to 10 min of exposure significantly improves the advanced parameters of mobility, namely, curvilinear velocity (VCL), linear velocity (VSL), the mean velocity of the path (VAP), and the amplitude of the displacement (ALH). The effect of the increase in the VCL decreased the linearity (LIN), the mean line (STR), and the mean wobble (WOB). Oregano oil at 5 min had no significant effect on the DNA fragmentation index (DFI) and sperm decondensation index (SDI). However, at 10 min, it had a significant effect on both DFI and SDI. The analysis of our results showed that this plant oil rich in terpenoids and phenolic antioxidants could be a quite good in vitro additive with high potential for the world of medically assisted reproduction

The Effect of Supplementation with Some Essential Oils on the Mobility and the Vitality of Human Sperm

The objective of this work is to study the improvement effect of some essential oils of sage (Salvia officinalis), oregano (Origanum vulgare), and eucalyptus (eucalyptus globulus) on the physiological parameters characterizing the quality of human sperm (mobility and vitality). We find natural biomolecules to improve sperm quality to increase the chances of success of very low in vitro fertilization (IVF) that stagnate around 20%. Sperm samples were mixed with different concentrations of essential oils. The effect of these essential oils on the motility and vitality of spermatozoa has been analyzed. The mobility was determined using a Computer Assisted Sperm Analysis (CASA). In the other side, the evaluation of sperm vitality was performed by staining eosin 2% and the microscopic examination is carried out via optical microscope. A drop of sperm will be mixed with a drop of eosin solution 2%, spread between the slip and coverslip, then allowed to air dry, and examined under a microscope. A significant improvement in the mobility and vitality of human spermatozoa has been noted with oregano. Eucalyptus after 10 min of exposure also significantly improves the mobility and vitality of the spermatozoa. Sage does not improve mobility for these incubation times but significantly improves vitality