Establishment of Blastocystis hominis in-vitro Culture Using Fecal Samples from Infants in Slum Area of Mirpur, Dhaka, Bangladesh.

Introduction: Blastocystis hominis (B. hominis) is an obligate anaerobic protozoan found in the human large intestine, and is the most common eukaryotic organism reported in human fecal samples. Method: Multiple stool samples from 460 children (53.9% male and 46.07% female) were collected and examined for the presence of Blastocystis hominis in Parasitology Laboratory of International Centre for Diarrhoeal Diseases Research, Bangladesh during the period of 9th January to 28th December, 2011. Among them, 255 were diarrheal patients (56.47% male and 43.53% female). Direct microscopy was done for each of the samples and each sample was cultured in vitro for 48 hours and observed again for the presence of the pathogen. Th e aim of the study was to develop a sustainable technique to identify the pathogen. Results: In culture, several morphological forms were observed. Th rough microscopy, various morphological forms were clearly observed. Within 5679 tested samples, 795 samples (0.14%) were positive for B. hominis. As multiple forms were observed in the same sample, the most prevalent was cyst (0.125%) whereas least prevalent was granular (0.0072%). Th e highest percentage for all the morphological forms was observed in age group 25-36 months. In direct microscopy from fresh samples, children from 37-48 months showed the highest percentage (22.9%) of infection (p=0.000). In culture, the same age group showed the most infection rate (p=0.000). Among the diff erent morphological forms observed in culture, the highest prevalence of cyst was in age group 37-48 months (p=0.000). Th e highest prevalence of vacuolar form(5.7%) was observed in the same age group (p=0.015). In contrast, the amoeboid forms were mostly observed in children of 25-36 months (p=0.002).Th e children aged in between 37 to 48 months are at the most risk of the infection. Conclusion: Th e sensitivity of direct microscopy was found only 38.46% in respect to in-vitro culture which strongly suggests that in-vitro culture is the gold standard for the diagnosis of this parasite.

Degradation of Resin Composites in a Simulated Deep Cavity

The aim of this study was to analyze the sorption and solubility of a nanofilled (Filtek Z350) and a midifilled (Filtek P60) resin composite in oral environment-like substances, in a simulated deep cavity. A cylindrical cavity prepared in a bovine incisor root was incrementally filled with resin composites. The obtained resin composite cylinders were cut perpendicularly to the axis to obtain 1-mm-thick discs that were divided into fifteen groups (n=5) according to depth (1, 2, 3, 4 and 5 mm) and immersion media (distilled water - DW, artificial saliva - AS and lactic acid - LA). The sorption and solubility were calculated based on ISO 4049:2000. Additionally, the degree of conversion (DC%) was calculated by FT-IR spectroscopy. Data were analyzed using multifactor analysis of variance (MANOVA) followed by Tukey's HSD post-hoc test and linear regression analysis (a=0.05). The DC% was higher for the midifilled resin composite and was negatively influenced by cavity depth (p<0.05). The nanofilled resin composite presented higher sorption and solubility than did the midifilled (p<0.05). The immersion media influenced the sorption and the solubility as follows: LA>AS>DW, (p<0.05). Both phenomena were influenced by cavity depth, with the sorption and solubility increasing from 1 to 5 mm (p<0.05). The degradation of resin composite restorations may be greater in the deepest regions of class II restorations when the composite is exposed to organic acids present in the oral biofilm (lactic acid).

O objetivo deste estudo foi analisar a absorção e a solubilidade de uma resina composta nanopartículada (Filtek Z350) e de uma híbrida (Filtek P60) em substâncias simuladoras do ambiente oral em um modelo de cavidade profunda. Uma cavidade cilíndrica, construída em uma raiz de um incisivo bovino, foi preenchida incrementalmente com as resinas compostas. Os cilindros obtidos foram seccionados perpendicularmente ao eixo para obtenção de discos com 1 mm de espessura, que foram divididos em 15 grupos (n=5) de acordo com a profundidade (1, 2, 3, 4 e 5 mm) e o meio de imersão (água destilada - AD, saliva artificial - SA e ácido lático - AL). A absorção e a solubilidade foram calculadas com base na norma ISO 4049:2000(E). Adicionalmente, o grau de conversão (GC%) foi calculado através de espectroscopia FT-IR. Os dados obtidos foram analisados usando análise de variância multifatorial, teste de Tukey HSD e análise de regressão linear (a=0,05). A resina composta híbrida apresentou o maior grau de conversão, sendo este influenciado pela profundidade da cavidade (p<0,05). A resina composta nanopartículada apresentou maiores valores de absorção e solubilidade (p<0,05). O meio de imersão influenciou a absorção e a solubilidade: AL>SA>AD (p<0,05). Ambos os fenômenos foram influenciados pela profundidade da cavidade, com os valores aumentando de 1 para 5 mm (p<0,05). A degradação de restaurações de resinas compostas pode ser maior em regiões profundas de restaurações classe II e quando o material é exposto à ácidos orgânicos produzidos pelo biofilme oral (ácido lático).

[ species of intestinal parasites and their prevalence in children in elementary educational level in rural of Damascus-Kalamoon-Syria]

708 stool samples were collected from children in elementary educational level in Kalamoon area [Nabek-Deratiah-Kara], in rural of Damascus-Syria, and were examined. That was done between November-January of 2009-2010. The aim of this study is to find out the species of intestinal parasites spread in this area [This study is done as the first time in this area]. We found that 125 samples contain parasite, which equals to 17.65%, and 65 samples contain human pathogenic parasites 52%. The pathogenic parasites according to their prevalence: Giardia lambilia 6.07%, Chilomastix mesnili 0.14% and Candida SP 2.96%. While the nonpathogenic parasites prevalence: Entamoeba coli 1.27%, Blastocystis hominis 8.05%, Endolimax nana 0.98%. It is worthy to mention that no case of intestinal worm ova such as Ascaris lumbricoides and Entamoeba histolytica has been diagnosed in our study

Use of multiplex PCR in the differential detection of Entamoeba histolytica and/or Entamoeba dispar in comparison to microscopic examination

Intestinal amoebiasis, caused by E. histolytica, is a major public health problem in developing countries. E. dispar is morphologically identical to E. histolytica, but is considered a commensal protozoan. Differential identification of E. histolytica and/or E. dispar is important for treatment and epidemiological studies of intestinal amoebiasis. To evaluate multiplex PCR in the differential detection of E. histolytica and/or E. dispar in comparison to conventional microscopic stool examination. The present study was performed on 100 individuals. They were divided into symptomatic and asymptomatic groups according to the presence or absence of symptoms and signs of intestinal amoebiasis. Stool samples were collected and subjected to microscopic examination of fresh preparations of direct wet mount and formalin diethyl ether concentrates, and of permanent preparations stained by trichrome, in addition to molecular assay using multiplex PCR. Percentage detection of Entamoeba species was highest by multiplex PCR [36%] and lowest by direct wet mount [25%]. Multiplex PCR detected E. histolytica in 25%, E. dispar in 41.7% and mixed infection in 33.3% of cases. The sensitivity of PCR was 96.4% compared with the trichrome stain [75%]. Multiplex PCR was more sensitive than the conventional microscopic techniques in the differential detection of E. histolytica and/or E. dispar, at the same time and in a single PCR round

Evaluation of some micronutrients, antioxidant biomarkers and total antioxidant capacity in human giardiasis

Giardia lamblia, a common parasite of the human small bowel, is responsible for diarrhea, nutritional deficiencies and production of free radicals. Micronutrients regulate many physiological functions and participate in antioxidant defense. Changes in their serum levels occur in several gastrointestinal disorders. The present study aimed to investigate the homeostasis of some micronutrients and antioxidant defense in human giardiasis. The study was designed to compare serum levels of some micronutrients and antioxidant status in 30 patients with symptomatic giardiasis and 30 age- and sex-matched controls. Giardia trophozoites and/or cysts were detected in the stool samples of patients using saline and iodine wet mount preparations and formol ether technique. Levels of magnesium, iron, manganese, copper and chromium were measured using atomic absorption spectrophotometer. Vitamin E level was determined as well as total antioxidant capacity [TAC] and serum levels of the antioxidant biomarkers; albumin, total bilirubin and uric acid were evaluated. Giardia-infected patients displayed significantly lower levels of magnesium, iron, manganese, chromium and vitamin E and higher levels of copper and uric acid compared to controls. TAC, albumin and bilirubin in Giardia-infected patients were not significantly different from the controls. The pathophysiological changes in giardiasis alter the homeostasis of some micronutrients. Uric acid constitutes an important antioxidant defense in human giardiasis that may compensate for the deficiency of some antioxidant micronutrients. Prevention and treatment of giardiasis may have a positive impact on the micronutrient status and on the general health of the population

Factors affecting the occurrence of hydrothorax in Egyptian patients with decompensated liver disease

It is well known that in cirrhotic patients, a large volume of ascetic fluid is generally well tolerated due to the capacitance of the peritoneal cavity. On the other hand, even modest volumes of pleural fluid can cause significant respiratory symptoms, including dyspnea and chest pain. Therefore, although infrequent, hepatic hydrothorax may represent a major clinical problem in the management of patients with portal hypertension. This work search for factors affecting the occurrence of hepatic hydrothorax in Egyptian patients with decompensated liver disease A total of 40 patients selected from Al Azhar University Hospitals with decompensated liver disease and ascites were included in this study divided into two groups 20 patients have ascites without pleural effusion were selected to be included in the study as a group [I], another a 20 patients have ascites with pleural effusion were selected also, to be included in the study as a group [II].All patients were subjected to the following:-Careful history taking, Careful clinical examination, laboratory investigations, including, complete urine and stool analysis, complete blood picture and, Liver function tests, Renal function tests, ascitic fluid and pleural fluid analysis for, physical, chemical, Cytological examinations. Abdominal Ultrasonography, Chest X ray, Results showed that the mean of BMI and ascitic fluid LDH are higher in patients with pleural effusion than in patients without pleural effusion. In contrast, the mean of alkaline phosphatase, ascitic fluid glucose and ascitic fluid RBCs are lower in patients with pleural effusion than in patients without pleural effusion but these findings are not enough to explain the occurrence of hepatic hydrothorax in those patients and these factors still obscure so more studies are needed to detect these factors

Detection of Giardia lamblia antigen in stool specimens using enzyme-linked immunosorbent assay

The aim of this study in Iraq was to determine the sensitivity and specificity of a commercial ELISA test for detection of Giardia lamblia antigen in stool. Of 84 stool samples from children in Duhok governorate, 42 were positive and 42 negative for G. lamblia or other parasites by direct and indirect microscopic examination. The sensitivity of the ELISA test for detection of G. lamblia versus microscopy was 76.4% and the specificity was 100%. We recommend using ELISA in epidemiological surveys in Iraq and to confirm the diagnosis in patients with typical clinical symptoms of giardiasis but negative results by direct microscopy

Clinical and laboratory predictors of acute bacterial diarrhea

Diarrhea is one of the most common childhood illnesses worldwide and exerts major financial impact. The presence of gross blood or an inflammatory response in the stool [leukocytes or lactoferrin] significantly increases the chance for isolation of invasive enteric bacteria. Improved knowledge of the microbiologic etiology of childhood diarrhea could help clinicians make appropriate diagnostic and therapeutic decisions and diminish the burden of these illnesses. The aim of this work is to determine the predictive utility of certain clinical and stool parameters in diagnosing bacterial diarrhea. A prospective one year study was conducted on 142 patients, below 5 years of age, with acute diarrhea, in Babylon Maternity and Children Teaching Hospital [Outpatient and Emergency department] during the months of January to December 2007. The positive stool culture was yield in 64 patients [45.07%]. The isolated pathogens were Escherichia coli in 44 patients [68.7%], Shigella spp. in 14 patients [21.6%], and Slamonella spp. in 6 patients [9.3%]. The best predictive variable for a stool culture positive for a bacterial pathogen was the presence of fecal leucocytes, with a sensitivity of 92%, a specificity of 75%, and positive and negative predictive values of 75% and 92%, respectively, with odd ratio of 36.6. The next predictive variable was the presence of both fecal leucocytes and fecal blood with a sensitivity of 88%, a specificity of 82%, and positive and negative predictive values of 76% and 91%, respectively, with odd ratio of 36.1. The best historical factors for predictive accuracies were abrupt onset, fever >/= 38[degree sign]C, abdominal pain, frequent bowel motions and no vomiting before the onset of diarrhea. Examination of stool for fecal leukocytes and fecal blood is a rapid, reliable, and inexpensive way to be the best method of predicting a positive stool culture, while detecting a blood in the stool only had high specificity with low sensitivity

Preliminary evaluation of the role of faecal calprotectin in the diagnosis of ulcerative colitis among Egyptian patients

Calprotectin, a member of the Ca2+-binding S100 family of proteins, makes up about 5% of the total protein content of the neutrophil. It is released upon activation and degranulation of neutrophils and correlates strongly with 111-indium-labeled leukocyte excretion. It was investigated as a promising tool in differentiating between Irritable Bowel syndrome [IBS] and active Inflammatory Bowel Disease [IBD]. Is to compare faecal Calprotectin in patients known to have ulcerative colitis with normal healthy controls and to investigate possible correlation of Calprotectin with disease activity on clinical, laboratory and pathological bases. Forty patients known to have UC were assessed. 19 were excluded: 10 due to non-steroidal anti-inflammatory [NSAID] intake, 2 due to pregnancy and 7 due to disease quiescence. So, 21 patients with active disease were studied, 7 males [33.3%] and 14 females [66.7%], mean age 37.5 [ +/- 16.0] years. Ten healthy controls [8 females and 2 males], mean age 30.9 [ +/- 16.1] years were included. Patients underwent clinical evaluation, determination of blood Erythrocyte sedimentation rate [ESR], C-reactive protein [CRP] and faecal Calprotectin. Colonoscopy was done to confirm diagnosis, estimate disease extent and obtain colonoscopic biopsy specimens for histological grading of activity. An overall scoring of disease activity was done using the Mayo score. Faecal Calprotectin was significantly elevated among patients [mean: 12.6 micro gm/gm stools [ +/- 3.2] in comparison to controls [9.4 micro gm/gm stools [ +/- 2.6], [p 0.01]. At a cut off of l0.3micro gm/gm stools it has a sensitivity of 86%.specificity of 70% p=0.004, positive predictive value of 86% and a negative predictive value of 70%. No correlation was found between faecal Calprotectin and ESR, histopathology and Mayo score. Calprotectin was significantly higher in cases with left sided colitis [l4.1 +/- 2.71gm/gm stools] than those with pnco1itis [11.8 +/- l.9 micro gm/gm stools], p 0.02.Faecal Calprotectin is a good test in differentiating Egyptian patients with ulcerative colitis from healthy controls. Thus, its use as a screening test may be helpful in the selection of cases for endoscopic examination. It lacks specific correlation with the severity of ulcerative colitis. This leaves endoscopy and histopathologic examinations as the main diagnostic tools. Larger scale studies on Egyptian patients are strongly recommended with special reference to the local mucosal permeability and immune milieu of the Egyptian population

Frequency of Cryptosporidium and risk factors related to cryptosporidiosis in under 5-year old hospitalized children due to diarrhea

Recently Cryptosporidium has gained much attention as clinically human pathogen in immunocompromised cases and young children. This study investigated frequency and risk factors related to cryptosporidiosis in under 5-year old children. Stools were examined by ELISA method to detect Cryptosporidium surface antigen [CSA] using Remel Prospect Cryptosporidium [monoclonal] Microplate Assay. Stool samples were collected from children at admission and were kept at 70[degree sign] C until examination. Data were collected by a standard questionnaire and analysed in Epi info 2002 software. Of 171 children, 8[5%] were infected with Cryptosporidium. Most of the cases [6 cases] aged 2-12 months. Boys were more frequently infected than girls [p<0.05]. Meanwhile, cryptosporidiosis was associated with less breast-feeding and lower birth weight [p<0.05]. Results revealed that the frequency of cryptosporidiosis was similar to other parts of the world. Similarly, lower birth weight, less breast-feeding and male gender were associated with a higher frequency of cryptosporidiosis

Copro-antigen detection versus direct methods for the diagnosis of Giardia Lamblia in patients from the national liver institute

In this study a commercially available immunoenzymatic assay to detect G. lamblia specific copro-antigen was evaluated. A total of 90 stool samples were tested. Diagnosis of giardiasis by ELISA for copro-Ag detection was positive in 46 [51.1%] patients whereas by direct stool analysis after formol ether concentration G. lamblia was detected in 38 [42.2%] patients only. ELISA technique for detection of Giardia copro-antigen had a sensitivity of 97.3% and a specificity of 82.6% with PPV of 80.4% and a NPV of 97.7%

[Frequency of Campylobacter jejuni in stool samples of patients with bloody diarrhea]

Compylobacter jejui is an important pathogen in developing diarrhea, ufortunatley it was not mostly searched in ordinary diagnostic laboratories using selective C. jejuni medium and many diarrhea due to this bacterium remained undiagnosed. The aim of this study was to evaluate frequency of C. jejuni in stool samples of patients with bloody diarrhea. This is a descriptive cross-sectional study, in which frequency of C. jejuni was evaluated among 90 patients with diarrhea attending to Azad University hospitals between 2007 and 2008. Stool samples were cultured in skirrow's medium and incubated in 42°C for 24-48 hours and bacterial colonies were assessed biochemically. Ninety patients with mean age of 13.08 years [range age of 4-80 years] were studied. 24 [26.7%] were male and 66 [73.3%] female [p<0.05]. Of 90 patients, in 7 cases [7.8%] C. jejuni was obtained. Severe abdominal pain in C. jejuni positive cases was higher then C. jejuni negative subjects [28.6%] in C. jejuni positive patients versus 7.2% of C. jejuni negative cases, p<0.05]. Because of importance of C. jejuni in bloody diarrhea and bacillary dysenteries, it is recommended the use of campylobacter selective medium such as skirrow's or campybap in examination of diarrheic stools

Increased isolation of Vibrio cholerae O1 serotype Inaba over serotype Ogawa in Pakistan

Although the predominant Vibrio cholerae serotype in Pakistan is Ogawa and serotype Inaba is rare, there has been a significant increase in the isolation of Inaba in our referral laboratory in Karachi. This paper reports this observation and further analysis of previous cholera data from 1993 to 2005 to assess the trend of occurrence and resistance pattern of V. cholerae strains. From January to September 2005, 245/3292 [7.4%] specimens yielded growth of V. cholerae. Of these, 243 were serotype Inaba, outnumbering serotype Ogawa. This recent Inaba strain is 100% resistant to cotrimoxazole, 3% resistant to chloramphenicol and not resistant to ampicillin, tetracycline and ofloxacin. This sensitivity pattern is almost similar to that of the previous predominant serotype Ogawa

Factors associated with acute diarrhoea in children in Dhahira, Oman: a hospital-based study

To elucidate the bacterial etiology of childhood diarrhoea in Dhahira, 856 children < 12 years admitted for diarrhoea to Ibri Regional Referral Hospital from 2000 to 2002 were studied. The mean age was 2.4 [SD 2.3] years; the majority [92.9%] were < 5 years. Bacterial etiology was found in 15.2% of cases; 10.6% due to Shigella and 2.1% to Salmonella. Sh. sonnei was the commonest Shigella serogroup isolated. Salmonella infection was significantly associated with cramps, while Shigella infection was associated with fever, bloody stools and cramps. Antibiotics were prescribed in 36.2% of cases and the resistance to the common antibiotics tested was low

Shigella and Salmonella serogroups and their antibiotic susceptibility patterns in Ethiopia

In this study, the serogroup and susceptibility patterns of Shigella and Salmonella spp. isolated from stool cultures were determined using standard laboratory procedures. Among the 76 Shigella isolates serogroup B [Sh. flexeneri] was the most prevalent species [54.0%] and among the 37 Salmonella strains serogroup B was also the most prevalent [81.1%]. Antibiograms of Shigella and Salmonella spp. showed 100% resistance to erythromycin and high resistance rates [>/= 75%] to ampicillin, cephalothin, chloramphenicol and tetracycline. Salmonella spp. Also had high resistance to gentamicin, sulphonamide, and trimethoprim- sulfamethoxazole. Shigella were susceptible to gentamicin [100%] and nalidixic acid [97.3%] and Shigella and Salmonella were 100.0% susceptible to norfloxacin

Frequency of Escherichia coli O157: H7 in children with diarrhoea in Zahedan, Islamic Republic of Iran

Escherichia coli O157:H7 has been implicated in diarrhoea, dysentery, haemorrhagic colitis and uraemic haemolytic syndrome but is not routinely cultured in laboratories. We tested 322 samples from children referred with diarrhoea to a clinic in Zahedan, Islamic Republic of Iran. There were 21 sorbitol-negative E. coli isolated; serotyping revealed 4 strains positive for O157, out of which only 2 strains were identified as E. coli O157:H7. Both cases were boys, aged 1.5 and 4 years respectively. Antibiotic sensitivity testing found the strains were sensitive for amikacin, gentamicin, nitrofurantoin and tobramycin and resistant to chloramphenicol, cefalexin, cefalothin and co-trimoxazole

Intestinal parasitosis and use of untreated wastewater for agriculture in Settat, Morocco

An evaluation was made of the risk of contamination by faecal parasites in the population from the use of wastewater in agriculture. The study was based in 3 rural clusters in the northern region of Settat city, Morocco: 2 rural clusters exposed to untreated wastewater in agriculture and 1 not exposed. The sample comprised 333 people aged 3-60+ years: 214 exposed and 119 non-exposed. Stool samples were collected and examined for intestinal parasites. The results showed that the prevalence of intestinal parasitosis [i.e. evidence of at least 1 type of parasite] in the exposed group was significantly higher that the non-exposed group [66.4% versus 31.9%; relative risk 2.1]. The household crowding index was also higher in the exposed group

Helicobacter pylori infection and hyperemesis gravidarum: possible causal relationship with evaluation of a new immunoassay in stool compared with standard diagnostic techniques

The aim of the present study was to investigate any possible association between infection with Helicobacter pylori [H. pylori] and hyperemesis gravidarum [HG]. Moreover; evaluation of different methods used in the diagnosis of H. pylori during pregnancy aiming to present a simple non-invasive and reliable method. 68 pregnant women with hyperemesis gravidarum and 72 control pregnant women were enrolled in the study. All participants were examined both for H. pylori serum immunoglobulin G antibodies [HpIgG Ab], showing chronic infection, and H. pylori stool antigens [HpSA], and showing active gastrointestinal colonization. Serologically positive H. pylori infection was detected in 59 [86.8%] subjects of the hyperemesis gravidarum group and in 32 [44.4%] of the controls [P<0.01]. HpSA was detected in 45.6% of patients with hyperemesis gravidarum, whereas only 5.6% of patients in the control group were positive for this specific antigen [P<0.001]. The new stool immunoassay test had a sensitivity of 96% [95% confidence interval 90.6% to 100%], specificity of 93% [85.1% to 99.5%], positive predictive value of 92%, and negative predictive value of 96%. In conclusion, this study supports the studies suggesting an association between H. pylori and HG. Infection with H. pylori should be kept in mind in cases of HG in pregnant women. The findings of the current study have, also, demonstrated that HpSA as a relatively simple, inexpensive and time saving noninvasive test is a reliable method for detection of active H. pylori infections in pregnant women with hyperemesis gravidarum. This stool immunoassay represents a new, accurate, and non-invasive method for H pylori infection that overcomes the limitations of existing tests

Hookworm infestation in children presenting with melena-case series

Hookworm infection is common in children and can present with symptoms of upper gastrointestinal bleeding and severe anemia. Ten children below 5 years presenting with melena and severe pallor were seen from December 2006 to May 2007 in the gastroenterology and hepatology department of children's hospital, Lahore. All patients had history of transfusion. Complete blood picture, eosinophil count with peripheral smear, stool complete examination for ova and cysts were performed in all cases, while upper and lower gastrointestinal endoscopies were performed in three patients to locate the source of bleeding. Stool routine examination in all these cases confirmed hook worm ova. These patients were managed with antihelmenthic and stool complete examination was done three days after the medicine. There was no mortality. Though upper gastrointestinal bleeding with hookworm infestation is very rare but in the developing countries it should be considered when other causes of upper gastrointestinal bleeding are ruled out