Showing papers in "Virology & Mycology in 2014"

Human Respiratory Coronaviruses Detected In Patients with Influenza-Like Illness in Arkansas, USA

Abstract: Acute respiratory viruses often result in significant morbidity and mortality. The potential impact of human respiratory coronavirus (CoV) infections was underestimated until the severe acute respiratory syndrome (SARSCoV) outbreak in 2003, which showed that new, highly pathogenic coronaviruses could be introduced to humans, highlighting the importance of monitoring the circulating coronaviruses. The use of sensitive molecular methods has contributed to the differential diagnosis of viruses circulating in humans. Our study aim was to investigate the molecular epidemiology of human CoV strains circulating in Arkansas, their genetic variability and their association with reported influenza-like symptoms. We analyzed 200 nasal swab samples, collected by the Arkansas Department of Health in 2010, for influenza diagnosis. All samples were from patients showing acute respiratory symptoms while testing negative for influenza. Samples were pre-screened, using a quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) multiprobe for coronavirus, and subjected to confirmatory pancoronavirus and/or strain-specific reverse transcriptase (RT)-PCR followed by sequence analysis. Seventy-nine samples (39.5%) were positive by qRTPCR and 35 samples (17.5%) were confirmed by conventional RT-PCR. Twenty-three of the confirmed samples (59%) were sequenced. The most frequent strain detected was HCoV-OC43-like followed by NL63-like; only one sample was positive for HCoV-229E and one for HCoV-HKU1. Feline-like CoV strains were detected in three samples, representing possible evidence of interspecies transmission or a new human strain. Seventeen percent of the coronavirus positive samples were also positive for other respiratory viruses, such as Respiratory Syncytial Virus (RSV), Parainfluenza 2 and 3, and Rhinovirus. Thus, HCoV-OC43, NL63, HKU1 and new feline-like strains were circulating in Arkansas in 2010. HCoV was the sole respiratory virus detected in 16% of the patients who showed acute respiratory symptoms with negative diagnoses for influenza virus.

A Clinico-Mycological Study of Superficial Mycoses from a Tertiary Care Hospital of a North Indian Town

Abstract: Introduction: Superficial mycoses refer to the diseases of skin and its appendages caused by fungi. Aims & Objectives: The present study was undertaken to find out the prevalence of different clinical types of Dermatomycoses including Dermatophytes, candida spp. and dermatomycotic mold infections of skin and its appendages and their etiological agents in and around Aligarh region of Northern India. Material and Methods: The study was conducted at the Department of Microbiology, JNMC,AMU, Aligarh, during the period of July 2011-July 2013. A total of 425 samples were collected including skin, nail and hair for a period of 5 years. Skin, nail scraping and clippings of infected hair from patients were collected. The samples were subjected to direct microscopy and culture. Results: Out of 425 samples, Dermatophytoses was manifested clinically more in the males between the age of 11-30 years. In our study, KOH positivity rate was 61.2% and culture positivity rate was 58.8%. 18.8% of samples were culture positive alone; while, 21.2% of specimens were positive on direct microscopy alone. Dermatophytoses was the commonest superficial fungal infection in 123 cases 28 (9.5%), followed by Dermatomycotic molds in 32 (7.2%) and candidiasis in 15 (3.5%). T.rubrum was the predominant pathogen followed by T. mentagrophytes. Conclusions: It is concluded that along with Dermatophytes, non-dermatophytic fungi are also emerging as an important cause of superficial mycoses.

Enhancing Seed Germination and Vigor of Chickpea by Using Potential and Effective Strains of Trichoderma Species

Abstract: In this study, potential and effective strains of Trichoderma such as T. harzianum (Th. Azad) and T. viride (01PP) have been investigated and their effect of pre sowing seed treatment on germination, seedling establishment, seedling dry weight and vigour in chickpea genotype (Radhey) was observed. The different pre sowing seed treatments showed different responses against all seven seed quality parameters. Chickpea seeds were treated with different concentrations of Trichoderma bioformulation such as 5%, 10%, 20% gm/kg seed followed by treatment with 0.2% Bavistin. As a result, the percentage of seed germination was found to be higher in T. harzianum (Th. azad) & Trichoderma viride (01PP) treated seeds with 5% bioformulation as compared to the other concentrations. Various attributes with their observations include seed germination (92% and 90%), root length (12.38 and 12.19 cm) shoot length (4.97 and 4.32 cm) seedling length (17.38 and 16.50 cm), dry weight (1.19 and 1.88 cm), vigour index I (5197.12 and 1485) and vigour index II (109.48 and 169.20). Among all treatments, control showed the poorest performance for all seven seed quality attributes.

A Comparison between ELISA and CIEP for Measuring Antibody Titres against Aleutian Mink Disease Virus

Abstract: Introduction: Enzyme-linked immunosorbent assays (ELISA) is used for the identification of mink with low antibody titre against the Aleutian mink disease virus (AMDV) who can tolerate the infection. There is no published information on the merit of ELISA for measuring antibody titre. An ELISA was compared with the counterimmunoelectrophoresis (CIEP) for quantifying antibodies against AMDV. Materials and methods: In experiment 1 (EX1), 41 black mink were inoculated with various amounts of a local strain of AMDV and were euthanized on day 218 post-inoculation (pi). In experiment 2 (EX2), 262 black mink were inoculated with a single doses of the same virus and euthanized 16 to 71 weeks pi. Antibodies to AMDV were quantified by CIEP using 10 two-fold serial dilutions of plasma, and serum samples were tested by the ELISA. Total serum proteins, albumin and globulins were measured in EX1. Results and discussion: All mink were tested positive for AMDV infection by PCR in both experiments. There were moderate concordances between CIEP and ELISA results (Spearman correlation of 0.77 and 0.71 in EX1 and EX2, respectively). Each ELISA class spanned a wide range of antibody titres in both experiments. Antibody titres of lower than 32 were not associated with serum proteins while greater titers were positively associated with total serum proteins and globulins (P<0.01) and negatively associated with albumin and albumin:globulin ratio (P<0.01). Regressions of serum protein measurements on ELISA classes were linear and in the same directions as, although weaker than, those on CIEP. Conclusion: Although ELISA was not able to accurately measure anti-AMDV antibody titre, in the absence of any other practical method, it is a useful tool for ranking mink for anti-AMDV antibody titres.

Natural Antivirals against Human Viruses

Abstract: Over the past two decades there has been intense study of antivirals from plants that have antiviral activity against viruses affecting plant and animal kingdoms. This volume of Virology and Mycology has a report by Awasthi et al. [1] on the antiviral activity against plant mosaic viruses affecting commercial important bean plants. A number of viruses that can cause diseases in humans can be either neutralized or its replication can be inhibited. There has been an explosion of research in the antiviral activity in recent years against human diseases caused by pathogenic viruses, which are known to affect millions of people across the globe and cause significant morbidity and mortality. As a champion of evidence based natural products of preventive and therapeutic agents, I would like to dedicate this editorial to those who have labored on researching natural products against all odds, often experiencing a hostile and elitist opposition to the discovery of natural products. The challenges to translating the findings of natural products are many and this editorial will conclude with steps one needs to take in order to obtain regulatory approval from agencies like the FDA and the EU regulatory group. Very few of the newly found compounds are anywhere close to finding human use. The one Secomet V (the active agent of which is Fulvic acid) that has reached human observational trial stage in HIV patients reported by Van Rensburg et al. in 2010 is a precursor to more proper trials, which have been presumably underway [2]. But the promise of Secomet V, a broad spectrum natural product, first reported by Kotwal et al., [3] takes exceedingly long periods to go from bench (discovery and laboratory testing) to bedside (administered in patients). This passage of over a decade is due to the underfunding of natural products evidence based research in the western world and the low impact they make in the publication arena in top medical journals like Nature Medicine, who look down upon the struggling researchers from the natural products field. Then there are those like pomegranate, found and consumed universally and known to have health benefit and documented to have broad spectrum activity (Table 1) against enveloped viruses like HIV, Influenza, Herpes and non-enveloped food borne viruses Noroviruses, have yet to establish therapeutic levels for human use, making them less practical for realization of their valuable potential. There are a number of naturally found compounds that have been identified against medically important viruses, which together contribute to annual infections of over billion people around the globe, and have been grouped here and summarized in tables. They include the antivirals to human hepatitis viruses (Table 2), viruses causing respiratory infections (Table 3), picornaviruses which cause a number of diseases (Table 4), human herpes viruses (Table 5) and Human Immunodeficiency virus (Table 5). In addition to the medically important viruses included in the tables there are other viruses of pediatric importance viz the Rota virus which causes diarrhea leading to severe dehydration and significant infant deaths also has a natural antiviral that has been identified [4]. With such a wide range of a repertoire of identified antivirals from natural sources like plants, what are the possibilities of how they could find human use? As in the case of Fulvic acid and pomegranate juice, they could be therapeutically used to neutralize pathogenic viruses like HIV, Influenza in human tissues and thus described as the enveloped virus neutralizing compounds (EVNCs). The advantage of such compounds is that they are not susceptible to the vagaries of mutations and drug resistance due to their mode of action, which is not dependent on the changes in amino acid sequence due to mutations [5]. EVNCs could be used in prophylaxis, as vaccine generating agents and maybe even as microbicides. Although microbicides have yet to realize their promise in providing women control over their sexual activity while ensuring that transmission is blocked. Finally, what steps will have to be taken following identification of antiviral activity? Guidelines are summarized in Table 7. The science preceding the translation has to be robust, if not the long path to human use can be costly waste of time and resources. It will be evident from the work in the field of natural antivirals that scientists from China are the leaders in the field and hopefully will continue to receive support to take on the long journey to finding their way into human use. A decade ago, in an editorial, I had envisioned that HIV could be eradicated by 2050 [49]. But amongst the medicines that could achieve that goal, I had an expectation that natural medicine would play a significant role. After reviewing the

Immune Thrombocytopenia Following Influenza Virus Infection and Influenza Vaccine Administration

Abstract: Immune thrombocytopenia (ITP) may be induced by the reaction of autoantibodies with platelets. Although the precise mechanism remains to be elucidated, certain viral infections or vaccine administration can lead to ITP. However, ITP induced by influenza virus infection or influenza vaccine administration is infrequent and thus there have been few related reports. Here, we present a review of relevant case reports in the English and Japanese literature.

Spectrum of Superficial and Deep Fungal Isolates in Northern Pakistan

Abstract: Fungi are an important cause of superficial and deep infections in our population. Lack of expertise in proper identification and inadequate diagnostic facilities often lead to underreporting of these infections and thus underestimation of true disease burden. This study was conducted at Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi, Pakistan, from January 2011 through December 2013. Samples included specimen collected from superficial and deep tissues, respiratory tract specimen, blood, bone marrow and other body fluids. Skin (35.1%) and nail (10.2%) samples were the most common specimens from superficial body sites. Tissue specimens from various body organs and bronchoalveolar lavage fluid were the predominant specimens received for investigation of deep seated fungal infections, contributing 34.9% and 5.9% of the total specimens respectively. Yeasts were isolated from 75(22.6%) samples; different species of Candida accounted for majority of the isolates. Growth of molds was detected in 257(77.4%) samples with Aspergillus spp. accounting for 149 (44.9%) of the isolates. Among dermatophytes, Trichophyton interdigitale 13(3.9%) was the most common isolate. Moulds other than dermatophytes were also isolated from skin, hair and nail samples and Alternaria alternata (4.8%) was the most common non-dermatophyte isolated from these sites. Fungal infections and their spectrum varies considerably in different geographical locations and in all cases not responding to antibiotics and high risk groups, a possibility of fungal cause should be sought.

Detection of classical swine fever virus by a surface plasmon resonance assay

Abstract: Test sera from vaccinated pigs at different time-frame were used to assess the sensitivity and specificity of an optimized Surface Plasmon Resonance (SPR)-chip-based detection method. Biomolecular interaction between Classical Swine Fever Virus (CSFV) and serum antibody were investigated in relative time using Biacore SPR system. An amount of 8860.93 RU of CSFV in 10mM sodium acetate of pH 5.0 was fixed on CM5 dextran matrix sensor chip. Serum from vaccinated animals was allowed to run over the whole CSFV in triplicate. The relative response from serum of 5th week and 7th week old swine towards the immobilized CSFV reacted variably. The 10 fold diluted serum response unit of 94.24 ± 11.34 RU indicated a limited immune response at week 5. However in week 7, the highest response in the serum at same dilution demonstrated a 2 fold increase at 189.33 ± 2.57 RU. Regeneration with glycine-HCl at pH 2.0 enabled successful baseline reversion after each analysis. The herein established, whole virus immobilized SPR-chip could serve as a prototype for a rapid and sensitive CSFV diagnosis assay.

Mushroom Insoluble Non-Starch Polysaccharides (MINSP) Attenuate Ochratoxin A-Oxidative Stress and Hepato-Renal Dysfunctions in Rat

Abstract: The aim of the present study was to investigate the effect of Mushroom Insoluble Non-Starch Polysaccharides (MINSP) on Ochratoxin A-induced hepato-renal damage in rat. MINSP (75 and 150 mg/kg) administered daily for 15 days in concomitantly with Ochratoxin A (OA) (1.7 mg/Kg), Intraperitoneal (i.p). Normal groups treated with MINSP showed significant decrease in serum levels of γ-Glutamyle Transferase (GGT), Aminotransferases (ALT & AST), creatinine, uric acid, total antioxidant capacity (TAC), Tumor Necrosis Factor Alpha (TNF-α ) and Carcinoembryonic Antigen (CEA). In liver and kidney homogenates there were significant decreases in Malonaldehyde (MDA), and Nitric Oxide (NO); while Superoxide Dismutase (SOD) content increased after treatment with MINSP. OA-treated rats showed significant elevation in serum ALT, AST, TNF-α, CEA, Uric acid , creatinine, MDA and NO and reduction of SOD level in hepatic and renal tissues. The combined treatment with MINSP+OA significantly ameliorated the tested parameters when compared with OA-treated group. It improved the antioxidant activity of the liver and kidney in dose-dependent manner. Histopathological examination revealed that MINSP administration protected hepatocytes and renal tissues from the damage induced by OA. Immunohistochemical staining of iNOS expression and DNA damaged area decreased in liver and kidney tissues after administration of MINSP. Conclusion: MINSP are fat replacer in processing low fat diet. MINSP represent a good functional food and have detoxifying effect against ochratoxicosis in rat.

Comparison of Surface-Plasmon-Resonance Biosensor Measurements and Western Blot Assay for Detection of GPE - Strain of Classical Swine Fever Virus by Using WH211 and WH303 Monoclonal Antibodies

Abstract: Monoclonal antibodies WH211 and WH303 are in routine laboratory use for detecting Classical Swine Fever Virus (CSFV) or its major envelope E2 protein. While E2 is recognized as the most immunogenic protein, it is also the most variable and often not recognized by specific monoclonal against this protein. The aim of this study was to compare the detection sensitivity of WH211 and WH303 antibodies for CSFV GPE- strain via two well-known assays, Surface Plasmon Resonance Biosensor and Western Blot Assays. Both WH211 and WH303 Abs which specifically known to detect E2 gene (gp55) of CSFV at different recognition sites (epitope) were used as ligands to detect the GPE- strain (analytes). GPE− strain showed interaction with monoclonal antibodies at highest dilution of 1:1000 (v/v) in SPR assay. At lowest dilution (1:10), the interaction of GPE− strain with immobilized monoclonal antibody WH211 showed more than two-fold increase (163.5 RU) than the interaction with monoclonal antibody WH303 (60.0 RU). This study documented profound preference for WH211 as the target for CSFV E2 gene by having higher sensitivity towards GPE− strain but with lesser affinity. E2 epitope of GPE− strain was found undetectable when blotted with WH303 at the investigated dilutions as compared to WH211. The findings indicated a 2500-fold higher SPR sensitivity in detection in comparison to Western Blot and the limit of detection of GPE− strain by Western Blot could not be achieved beyond 1:10 dilution of monoclonal antibodies. Therefore, SPR approach could overcome the risk of GPE− vaccine strain from being invisible for identification. Although, WH303 was unable to recognize this strain by Western Blot, both WH211 and WH303 were applicable as a sensitive detection ligand for GPE- strain of CSFV using SPR analysis.

The Quality of Ornamental Water within Shopping Malls in the United Arab Emirates

Abstract: Ornamental water fountains in shopping malls are mainly built for decorative purposes however; they might fail to meet the required standards set to ensure public health safety. Microbial contamination of indoor ornamental water could pose a serious potential risk to human health, particularly children and the elderly who could get infected by either direct contact or aerosol inhalation. Although the United Arab Emirates (UAE) is famous for its exquisite shopping malls that are routinely monitored and inspected for cleanliness, there are currently no unified standard regulations regarding microbial risk assessment for ornamental water. Cleaning and disinfection of the water is the responsibility of the mall management that resorts to a certified maintenance company which runs its own quality testing and follows its own guidelines. The objective of this study is to examine water quality from indoor ornamental fountains, and compare the different guidelines implemented with regards to microbial quality assurance and public safety. A time-series experimental design was utilized. Three main shopping malls within Abu Dhabi city were randomly selected for the study. Sampling and collection of biofilm swabs from water fountains and baseline sources were performed before and after cleaning in each mall. Sterilized containers where used to collect the water samples, and sterilized gloves where used throughout the whole process to prevent contaminating the water samples or the water feature. For examining the microbiological indicators of water quality, all samples were analyzed by the membrane filtration method and cultured on selective/differential media to determine total coliform count and other more specific bacteria like Staphylococcus aureus. The results of this study showed significant differences in the microbial profiles of ornamental water in the three malls with higher growth (CFU/100 ml) in 2 of the 3 of the decorative features examined. Moreover, the results showed differences between the results before and after the cleaning of the feature. Although, the average microbial growth from samples per each mall was within the limits for public health safety, this study emphasizes the important need for establishing proper guidelines for ornamental water within indoor malls in Abu Dhabi to ensure public safety at all times.

Testing of IgG and IgM against Rubella Virus in School Girls and PregnantWomen in Hanoi where an Outbreak had occurred

Abstract: Background: Congenital Rubella Syndrome (CRS) is preventable illness but stays frequent in developing countries where rubella vaccination is not incorporated in national program of vaccination like in Vietnam actually. Aims: This study aimed at obtaining baseline information on acquired immunization status of school girls as well as the rate of susceptibility and rubella infection in pregnant women around a rubella outbreak in Hanoi, Vietnam. Population and methods: During and after the rubella outbreak in 2011, sera from 136 school girls (aged from 14 to 17 years) and 140 pregnant women in Hanoi French hospital were tested for rubella-specific IgG and for rubella-specific IgM (only in pregnant women) by the technique of Electro-Chemiluminescence Immunoassay (ECLIA) (Roche Diagnostics) with cut-off value ≥ 10 IU/mL for IgG and ≥ 0.3 IU/mL for IgM. Results: Rubella-specific IgG antibodies was positive in 125/136 (91.9%) school girls and 119/140 (85%) pregnant women, with strong-to-extreme concentration in 86/136 (63.2%) school girls versus 19/140 (13.6%) in pregnant women (p<0.002). Rubella-specific IgG was negative (susceptible to rubella infection) in 21/140 (15%) and rubella-specific IgM antibodies was positive in 27.9% (39/140) pregnant women, of whom 38 (27.1%) a co-existence of rubella-specific IgG and IgM were documented. Conclusions: This preliminary, baseline per/post epidemic information showed a high rate of acquired immunization in female adolescents but also high rate of susceptibility and very high rate of rubella infection in pregnant women, warranting further comprehensive studies to determine robust, nationwide epidemiological data prior to establishing the national program of immunization against rubella and identify appropriate methods for CRS surveillance in Vietnam.

Biochemical and Immunogenetic Diagnostic Markers Less Commonly Used forPredicting the Efficacy of Chronic Hepatitis C Etiotropic Therapy

Abstract: Introduction: 1,7% of the population of Latvia in 2008 was HCV RNA positive, thus approximately 38 thousands required the CHC antiviral therapy. The objective of the study was to evaluate some biochemical and immunogenetic parameters allowing predicting the efficacy of the etiotropic therapy for CHC patients. Material and methods: Medical records of 213 CHC patients were analyzed. HCV RNA, anti-HCV, concentration of HA, GGT activity were detected and immunogenetic investigations were performed. Results and discussion: Evaluation of GGT activity showed increase in 46,81% of non-responders and in only 21,57% responders. HA level in responders was significantly lower than in non-responders. Correlation between the incidence of HLA class II gene alleles in CHC patients and the type of CHC therapy was found. The results of the study confirmed the hypothesis that some biochemical and immunogenetic parameters characterizing the condition of the macro organism had an essential role in the efficacy of the CHC therapy. Conclusions: The non-efficacy of the CHC etiotropic therapy was associated with increased GGT and HA levels upon the start of the therapy. The efficacy of PEG IFN+RBV combination therapy in comparison with the efficacy of Realdiron therapy was associated with different MHC HLA class II gene alleles.