Microbiol. Res., Volume 2, Issue 2 (August 2011) – 14 articles

Hepatitis C virus (HCV) has a major impact on public health. In spite of the progress made in the prevention of transfusion-transmitted infections over the last years, these still occur, especially in multi-transfused patients such as sickle cell anemia patients. Sickle cell disease (SCD) is highly prevalent in Eastern Saudi Arabia. Little is known about the prevalence of HCV in Saudi sickle cell disease patients. The present study aimed to assess HCV and HBV antigens, antibodies and viral genome among sickle cell anemia patients in a tertiary hospital in Eastern Saudi Arabia. Methods used included measurement of HCV antigen and antibodies using the novel HCV antigen/antibody combination assay, assessment of HCV core antigen and measurement of viral genome using standard commercial kits. Of the 138 sickle cell disease samples tested, 5 (3.6%) samples gave positive results. Their hemoglobin ranged between 7.8 and 10.1 g/dL, their erythrocyte count ranged between 3.1¥106 and 3.9¥106. Out of these 5 samples, 4 were also positive by the HCV Core Ag assay and by the HCV RNA PCR test (80%). None of the control group was positive. Seven patients were positive for HBs antibodies. One sample was positive for HBsAg, and this indicates chronic carrier state. Improving the testing for blood-borne infections such as HCV and HBV will result in better control of these infections in sickle cell disease patients which will inevitably lead to lower mortality and morbidity in this group of patients. Full article

The role of Chlamydia trachomatis (Ct) in everyday clinical practice is now on the increase because Ct infections are the most prevalent sexually transmitted bacterial infections worldwide. Ct can cause urethritis, cervicitis, pharyngitis, or epididymitis, although asymptomatic infections are quite common. Ct infection remains asymptomatic in approximately 50% of infected men and 70% of infected women, with risk for reproductive tract sequelae both in women and men. A proper early diagnosis and treatment is essential in order to prevent persistent consequences. An accurate comprehension of the pathology, diagnosis and treatment of this entity is essential for the urologist. We review the literature about the new findings in diagnosis and treatment of Ct infection in sexually active young men. Full article

Many systems are available for the isolation of anaerobic bacteria from clinical specimens. The jar system is the oldest and more adapted while the pouches are not popular with many investigators. The anaerobic chambers are expensive to maintain and technically inflexible. This study evaluated the efficacy of the Oxoid anaerobic jar and the GENbag pouches as anaerobic incubation systems. Anaerobic cultures were set up for 145 middle ear exudates and incubation was in the anaerobic jar, GENbag or a combination of both. The effect of specimen transport system and time lapse before culturing on the performance of the anaerobic systems were evaluated Ten genera of anaerobic bacteria were isolated with both systems (P > 0.05). Peptostreptococcus and Prevotella were isolated more frequently in Oxoid jar than in GENbag (P < 0.05) but both systems were not discriminatory for Clostridium, Propionibacterium and Veillonella. The use of GENbag as a backup to Oxoid jar increased isolation rate from 56.6% to 90.3% (P > 0.05). Type of transport media or vehicle did not affect the recovery of anaerobes adversely as did delay in processing of specimen. A careful application of a number of variables may improve isolation of anaerobes from clinical specimens Full article

Cystic echinococcosis (Hydatid disease) is a zoonotic disease caused by Echinococcus granulosus. The aim of this study, was to evaluate the scolicidal activity of methanolic extract of Zingiber officinale (Rosc.) family Zingibe - raceae, against protoscolices of hydatid cyst. Protoscolices were collected aseptically from sheep livers containing hydatid cyst and were exposed to different concentrations of ginger extract for various exposure times. Scolicidal activity of Z. officinale extract at concentration of 25 mg/mL was 25.6%, 39.1%, 56.7%, 83.7%, 98.1% and 100% after 10, 20, 30, 40, 50, and 60 min of exposure respectively. Scolicidal effect of this extract at concentration of 50 mg/mL was 52%, 85.8 %, 99.6% and 100% after 10, 20, 30 and 40 min of exposure respectively. Z. officinale extract at concentration of 100 mg/mL killed 76.5 %, 87% and 100 % of protoscolices after 10, 20 and 30 min respectivly. The results of this study showed that the methanolic extract of Z. officinale has high scolicidal activity and might be used as a natural scolicidal agent. Full article

We present here a case of cough, expectoration and fever for six days. After investigation by the doctor and epidemiologist, it was confirmed that this patient had a history of contact with a novel influenza virus. All the results of the real time PCR on novel Swine Influenza H1 were positive. Blood-gas assay PO₂50.6 mmHg, hypoxemia and computed tomography (CT) of lungs indicated patchy dense shadow scattered in both lungs in which the inflatable bronchial shadow was observed. A visible change in leakage around the shadow was seen. This was a serious case of infection from a novel influenza virus and the patient received systemic treatment: oseltamivir 75mg bid po, methylprednisolone 40mg qd iv, biapenem 0.6 q12h iv, and moxifloxacin 0.4qd po. On discharge from hospital, Swine influenza H1 was negative. Lungs showed apparent absorption of the inflammation. Respiratory failure had been corrected. Patients infected with a novel influenza virus should be given low-dose hormone and an antiviral drug as soon as possible after the infection is confirmed. Full article

P. falciparum malaria continues as the serial killer of over a million lives yearly, mainly for children in sub-Saharan Africa. For severe malaria, we are still on the quest for a prognostic marker of fatal outcome. We analysed the association between serum levels of Procalcitonin (PCT), a marker of septic inflammation, and clinical outcome in Senegalese patients admitted with confirmed cerebral malaria in the intensive care facility of Hopital Principal. A total of 98 patients living in the hypoendemic urban area of Dakar, Senegal, were enrolled during transmission seasons. Levels of PCT were compared between surviving vs the 26.5 % fatal cases in blood samples of the 3 days following hospitalisation. Mean PCT levels were elevated in patients with active infection, with a large range of values (0.1 to 280 nanog per mL), significantly higher on day 0 in fatal cases than in surviving (53.6 vs 27.3; P = 0.01). No exact individual threshold level could indicate occurrence of fatality, however mortality could be most accurately predicted by PCT level above 69 nanog per ML and there was a very clear different profile of evolution of PCT levels on the 3 days of observation decreasing early from day 1 in surviving patients (P < 10^–3), contrary to fatal cases. These results indicate that PCT kinetic rather than intrinsic level could be of use to predict a reduced risk of fatality in patient with cerebral malaria and could serve as potential predicting marker for severe malaria. Full article

The introduction of HBV vaccination in Italy has led to a decline in new HBV infections. Increasing immigration over recent years suggests a change in short-term epidemiology of HBV. The aim of this study was to assess the prevalence of HBV infection in the general population living in the catchment area of Legnano Hospital (Northern Italy). In the period 2007–2008, 22,758 inpatients and outpatients were examined for Hepatitis B surface antigen (HBsAg), of whom 1,654 (7.3%) were of foreign origin. Of the 488 patients who were positive for HBsAg (2.1%), 381 (1.8%) were Italian and 107 (6.5%) were born in other countries. In terms of age, the prevalence of HBsAg was significantly higher among non- Italians in every age group (other than those aged >60 and <11 years), and in many of the selected subgroups: the inpatients of some departments (35.4% vs 17.2%), pregnant women (5.3% vs 0.3%), blood donors (4.7% vs 0.1%), and hospital staff (6.4% vs 1.3%). Non- Italians were affected by 16.7% of acute infections and 24.3% of chronic infections; they also accounted for 42.6% of subjects with carrier state, 16.0% of patients with chronic hepatitis, and 12.2% of patients with cirrhosis. In our area, the overall prevalence of HBsAg among Italians is less than 2% (as expected following the introduction of HBV vaccination), but it is significantly higher among patients from areas highly endemic for HBV infection who represent a new reservoir for HBV infection. Full article

The present study is an investigation on the kinetics of dimethoate biodegradation and an estimation of residual dimethoate in bacterial culture by spectrophotometry. The methylene chloride extract of the culture medium was used for determination of dimethoate through its reaction with 1 chloro-2, 4 dinitrobenzene to produce methylamine whose absorbance at 505 nm gave an estimation of dimethoate content. The dimethoate standard curve follows Beer’s law at 505 nm with a slope of 0.0129 absorbance units per µg/mL. The regression equation relating concentration of dimethoate (x) with the absorbance is (y): y = 0.037 + 0.0129x. The amount of residual dimethoate after 7 days were 0, 4, 17, 28 and 29 µg/mL; the rate constants were 0.775, 0.305, 0.225, 0.167 and 0.127 each per day, and the efficiency of dimethoate degradation were 100%, 96%, 83%, 72% and 71%, for Bacillus licheniformis, Pseudomonas aeruginosa, Aeromonas hydrophila, Proteus mirabilis and Bacillus pumilus respectively. Dimethoate remediation could be attained through bacterial metabolism of the pesticide and colorimetric analysis might be useful in the estimation of dimethoate within a detection limit of 5–100 µg/mL. Full article

The frequency and range of long-term sequelae following acute Q fever infection remains controversial. This study aimed to assess a range of potential psychological and physical sequelae in a follow up study of a large outbreak of Q fever that occurred in Newport, Wales, in 2002. A nested case control study was undertaken six years after a point source outbreak of Q fever. We invited a cohort of 211 factory workers exposed to a point source of Q fever in 2002 to attend a follow up clinic in 2008. Cases, defined as those who had clinical symptoms and serological evidence of acute Q fever in 2002, were compared to controls, who worked in the same factory but were serologically negative at the time of the outbreak. At the follow up clinic blood was taken for Coxiella burnetii microimmunofluorescence and questionnaires were completed including the PHQ-9, Chalder Fatigue scale, and General Health Questionnaire. Results were obtained for 32 cases and 13 controls. Chalder Fatigue scores were significantly raised in the cases (independent samples t-test: P = 0.047). PHQ-9 and GHQ scores were not significantly raised in cases. However, post hoc cross sectional analysis indicated a relationship between Phase 2 IgG at follow up in 2008 and Chalder Fatigue scores (P = 0.004) and PHQ-9 scores (0.049). A longitudinal association was demonstrated between acute Q fever infection and chronic fatigue six years later. In crosssectional analysis a previously unreported relationship between depression scores (PHQ-9) and positive Q fever serology was also identified. Full article

Measles virus has no or indistinctive cytopathic effects (CPE) in cell couture system. Employment of some detecting methods like plaque assay or stereologic experiments, as a method of detecting of viral infection in the cells would be applicable. The aim of this study was investigating the early changes in quantitative parameters of measles virus infected Vero cells. Stereological methods using invariator, were applied for the first time to estimate cell and nucleus volume and cell surface of the infected Vero cell line with the measles virus.This method can be applied on other cultured cells.Vero cells grown in tissue culture plates for 48 hours at 36 ˚C were infected with 100TCID₅₀ of AiK strain of measles virus. Volume and surface of the infected Vero cells were studied at 4, 9 and 25 hours post infection along with uninfected control cells. The mean cell volume and surface of the cells infected with measles virus, increased ~87% and ~50%, respectively, 4 hours post-infection, as compared with the uninfected control. The nuclei did not show any differences. The mean parameters of infected cells in other time intervals showed no significant difference comparing with the control cells. Although there are other specific methods, stereology may be used as an integrated protocol to detect cytophatic changes of the measles virus infected cells early in the permissive cell culture system. Full article

Bioactivity guided isolation of dichloromethane extract of Alternaria alternata identified tenuazonic acid (1) as potentially active against Mycobacterium tuberculosis H37Rv, MIC at 250 μg/mL concentration. This active metabolite 1, was also evaluated for osmotic hemolysis using the erythrocyte as a model system. It was observed that this fungal metabolite showing antitubercular activity exhibited concentration dependent toxicity to human erythrocytes. Full article

This study brings the analysis of amino acid sequences of matrix protein (M1) from the influenza virus A (H1N1, H3N2 and H5N1) during 2007-2208. 741 sequences of M1 were compared, of them, H1N1 388; H3N2 251 and H5N1 102. Even though, the M1 is relatively conserved among the influenza A viruses, we found some variations in the M1 among the viruses, H1N1, H3N2 and H5N1. The nuclear localization signal at amino acid 101 to 105 is RKLKR for H1N1 and H3N2, but for H5N1 is KKLKR. All differences of amino acid in M1 of H1, H3 and H5 were listed. 80 sequences of M1 of H1N1 H3N2 and H5N1 were used for phylogenetic analysis. There is no reasontantment found in the M1 among these subtypes. Further study is needed to study the differences of the function of M1 among H1N1, H3N2 and H5N1. The M1 of H5N1 may contribute to the high pathogenesis to this virus. Full article

Alkaline soils commonly called as Usar soil having pH 7.5 to 11.0 is commonly found in regions having poor drainage and little percolation. These soils occupy vast tracts of barren lands in the northern parts of India. These areas are frequently visited by man and animals, however there are no reports on the occurrence of keratinophilic fungi from the Usar soils. The purpose of this research was to study the occurrence of keratinophilic fungi in the alkaline Usar soil. To investigate the occurrence of soil keratinophilic fungi from Usar soil, 120 samples were collected from five districts of Uttar Pradesh viz. Lucknow, Bareilly, Azamgarh, Balia and Pratapgarh. Keratinophilic fungi were isolated by the hair baiting techniques using human hair as keratin bait. The cultures were identified using macro- and micro morphological features. Eighty-tow strains of keratinophilic fungi were recovered from 120 (68.33%) soil samples. The isolated fungi were composed of ten species of five genera viz. Chrysosporium indicum (19.16%), Microsporum gypseum (12.5%), Chrysosporium tropicum (10.0%), Chrysosporium pannicola (7.5%), Trichophyton terrestre (5.83%), Chrysosporium lucknowense (4.16%), Chrysosporium state of Ctenomyces serratus (3.33%), Gymnascella dankailensis (3.33%), Gymnoascus reessii (1.66 %) and Trichophyton mentagrophytes (1.66 %). The presence of keratinophilic fungi in the Usar soils of Uttar Pradesh, India indicates that these fungi do occur at higher alkalinity. Full article

Avian influenza is a viral infection which affects mainly the respiratory system of birds. The H9N2 considered as low pathogenic avian influenza (LPAI) virus and continuously circulating in poultry flocks causing enormous economic losses to poultry industry of Pakistan. As these viruses have RNA genome and their RNA polymerase enzyme lacks proof reading activity which resulted in spontaneous mutation in surface glycoproteins (HA and NA) and reassortment of their genomic segments results in escape from host immune response produced by the vaccine. Efforts made for the isolation and identification of avian influenza virus from live desi and zoo birds of Lahore and performed antigenic characterization. The local vaccines although gives a little bit less titer when we raise the antisera against these vaccines but their antisera have more interaction with the local H9 subtype antigen so it gives better protective immune response. Infected chicken antisera are more reactive as compare to rabbit antisera. This shows that our isolates have highest similarity with the currently circulating viruses. These results guided us to devise a new control strategy against avian influenza viral infections. The antigenic characterization of these avian influenza isolates helped us to see the antigenic differences between the isolates of this study and H9 subtype avian influenza viruses used in vaccines. Therefore, this study clearly suggests that a new local H9 subtype avian influenza virus should be used as vaccinal candidate every year for the effective control of influenza viral infections of poultry. Full article