Giardia Lambia Research Articles

Identification and verification of α-11 giardin-interacting protein

To identify and verify the interacting protein of α-11 giardin, so as provide the experimental evidence for studies on the α-11 giardin function. The yeast two-hybrid cDNA library of the Giardia lambia C2 strain and the bait plasmid of α-11 giardin were constructed. All proteins interacting with α-11 giardin were screened using the yeast two-hybrid system. α-11 giardin and all screened potential interacting protein genes were constructed into pBiFc-Vc-155 and pBiFc-Vn-173 plasmids, and co-transfected into the breast cancer cell line MDA-MB-231. The interactions between α-11 giardin and interacting proteins were verified using bimolecular fluorescence complementation (BiFC). The yeast two-hybrid G. lambia cDNA library which was quantified at 2.715 × 107 colony-forming units (CFU) and the bait plasmid containing α-11 giardin gene without an autoactivation activity were constructed. Following two-round positive screening with the yeast two-hybrid system, two potential proteins interacting with α-11 giardin were screened, including eukaryotic translation initiation factor 5A (EIF5A), calmodulin-dependent protein kinase (CAMKL) and nicotinamide adenine dinucleotide phosphate-specific glutamate dehydrogenase (NADP-GDH), hypothetical protein 1 (GL50803_95880), hypothetical protein 2 (GL50803_87261) and a protein from Giardia canis virus. The α-11 giardin and EIF5A genes were transfected into the pBiFc-Vc-155 and pBiFc-Vn-173 plasmids using BiFC, and the recombinant plasmids pBiFc-Vc-155-α-11 and pBiFc-Vn-173-EIF5A were co-tranfected into MDA-MB-231 cells, which displayed green fluorescence under a microscope, indicating the interaction between α-11 giardin and EIF5A protein in cells. The yeast two-hybrid cDNA library of the G. lambia C2 strain has been successfully constructed, and six potential protein interacting with α-11 giardin have been identified, including EIF5A that interacts with α-11 giardin in cells.

Intestinal Parasites among Asymptomatic Food Handlers in a Tertiary Institution in Owo, Ondo State, Nigeria

ABSTRACT Incidence of intestinal parasites has been described as one of the most common problems of developing countries like Nigeria. This study was carried out to determine the possible presence of parasites amongst asymptomatic food handlers in tertiary institution in Owo. One hundred and twenty three (123) stool samples were randomly collected from food vendors in Achievers University, Owo. Also, questionnaires were administered and collated. The samples were transported to the University Parasitology Laboratory within 1 hour of production and examined microscopically using physiological saline and iodine direct mount and formal ether concentration techniques. Out of the 123 stool samples analyzed, 20 were infected with gastrointestinal parasitic organisms, giving a prevalence of 16.26%. Three different parasites were identified: Ascaris lumbricoides (12.20%), Giardia lambia (5.67%), and Entamoeba histolytica (3.25%). The prevalence with respect to gender showed that parasitic infestation was higher in females (9.76%) than in males (6.50%). The parasitic infestation was higher in age groups 36–45 years (7.32%), followed by age groups 18–25 years (5.69) and least in age group 26–35 years (3.25%). The parasitic infestation was higher among subjects who sell cooked food and snacks (6.50%), followed by those who sell cooked food only (4.88%), and lastly 2.44% among those that sell snacks and roasted meat respectively. In conclusion, this study confirmed the presence of intestinal parasitic infection among food handlers in Achievers University Owo, Ondo State. Good hygiene and safe food-handling should be exercised by food handlers.

Conserved Candidate Antigens and Nanoparticles to Develop Vaccine against Giardia intestinalis.

Giardia intestinalis (Giardia lambia, Giardia duodenalis) infections in humans may be asymptomatic or symptomatic and associated with diarrhea (without blood), abdominal cramps, bloating, flatulence, and weight loss. The protozoan Giardia is the third most common cause of diarrhea and death in children under five, preceded only by rotavirus and by Cryptosporidium parvum and C. hominis infections. Antimicrobial drugs, particularly 5-nitroimidazole (5-NIs), are used to treat giardiasis in humans. Immunologically naive or immunocompromised host are more vulnerable to Giardia infection, whereas a degree of resistance to this protozoan is present in humans living in endemic areas. This suggests that vaccination may be a potential and appropriate means to control this parasitic disease outbreak and protect the human population. This review discusses Giardia antigens related to vaccine development. Additionally, based on the latest development of nanoparticle technology, a combination of methods for future research and development is proposed for the design of the next generation of powerful immunogens and an effective vaccine against Giardia.

Efficacy of Capparis spinosa Linn Leaf and Fruit Extracts on Giardia Lambia Cysts In vitro

Aims: Giardia lamblia (G. lamblia) is one of the most common intestinal parasites worldwide. There are some side effects and the reports of parasite resistance to metronidazole as the first line treatment of giardiasis. Therefore, it is essential to discover an effect and safe drug to treat giardiasis.
 Methodology: In this study, the anti-parasitic activity of hydroalcoholic extracts of Capparis spinosa Linn (C. spinosa) leaves and fruits with different concentrations (4 to 0.125 mg/ml) was assessed against human isolates of the G. lamblia cysts and incubated at 37℃. After staining Giardia cysts with 0.1% eosin, the lethal percentage and 50% lethal concentration (LC50) of fruit and leave C. spinosa L extracts and metronidazole on G. lamblia cysts were calculated after 3, 6, 24 and 48 hours.
 Results: Anti-giardia activity of fruit and leaf extracts of C. spinosa was different between concentrations and time points (p<0.005). The lethal effect of both C. spinosa L extracts and metronidazole increased significantly in a concentration - and time–dependent response (P<0.0001). The highest lethal percentage of G. lamblia cysts was observed a concentration of 4 mg / ml of fruit (100%) and leaf (44%) extracts of C. spinosa L after 48 hours, respectively. LC50 values were 0.38 ± 0.02mg/ml for fruit extract, 2.32 ± 0.1 mg/ml for leaf extract and 0.53 ± 0.03 µg/ml for metronidazole after 48hours.
 Conclusion: C. spinosa can be effective in eliminating Giardia cyst of contaminated environments and water.

Identification of the NADP+ Structural Binding Site and Coenzyme Effect on the Fused G6PD::6PGL Protein from Giardia lamblia.

Giardia lambia is a flagellated protozoan parasite that lives in the small intestine and is the causal agent of giardiasis. It has been reported that G. lamblia exhibits glucose-6-phosphate dehydrogenase (G6PD), the first enzyme in the pentose phosphate pathway (PPP). Our group work demonstrated that the g6pd and 6pgl genes are present in the open frame that gives rise to the fused G6PD::6PGL protein; where the G6PD region is similar to the 3D structure of G6PD in Homo sapiens. The objective of the present work was to show the presence of the structural NADP+ binding site on the fused G6PD::6PGL protein and evaluate the effect of the NADP+ molecule on protein stability using biochemical and computational analysis. A protective effect was observed on the thermal inactivation, thermal stability, and trypsin digestions assays when the protein was incubated with NADP+. By molecular docking, we determined the possible structural-NADP+ binding site, which is located between the Rossmann fold of G6PD and 6PGL. Finally, molecular dynamic (MD) simulation was used to test the stability of this complex; it was determined that the presence of both NADP+ structural and cofactor increased the stability of the enzyme, which is in agreement with our experimental results.

Etiologies of diarrhea and drug susceptibility patterns of bacterial isolates among under-five year children in refugee camps in Gambella Region, Ethiopia: a case control study

BackgroundDespite substantial global effort and updated clinical management guidelines, diarrhea continues to be among leading worldwide causes of morbidity and mortality in children. Infectious diarrhea, the most common form of diarrhea causes substantial morbidity and mortality among children in developing countries, and the muddled use of antibiotics needs caution due to potential problems of drug-resistance. The aim of this study is to identify etiologies of diarrhea and drug susceptibility patterns of bacterial isolates in under-five children in refugee camps in Gambella Region, Ethiopia.MethodsAn institution- based matched case control study was conducted using a questionnaire-based interview from June to December 2017 in Pugnido and Teirkidi refugee camps. Stool samples were collected and parasites causing diarrhea were identified by wet mount microscopy. Conventional culture supplemented with API 20E identification kit was used to identify Salmonella and Shigella species. Antibiotic susceptibility of bacterial isolates was investigated by using the disk diffusion method. The association between etiologies and diarrhea was analyzed using McNemar test or Fisher exact test with 95% confidence interval at a level of significance of P < 0.05.ResultsThe overall prevalence of enteric pathogens were 55 (41.0%) in diarrhea cases and 18 (13.4%) in healthy controls. The detected etiologies include Giardia lambia (28), Shigella spp. (16), E. hystolyotica/dispar (13), Ascaris lumbricoides (10), Salmonella spp. (6), Cryptosporidium parvum (6), Hymenolepis nana (4) and Isospora belli (3). All isolates were sensitive to kanamycine and ceftazidime. The high resistance rate was observed against ampicillin (100%), amoxicillin (100%), erythromycin (52%), chloramphenicol (47.5%), tetracycline (40.5%), cotrimoxazole (34.8%) and amoxicillin-clavulanic acid (33%). The majorities of the isolates had a low rate of resistance to ciprofloxacin (8.7%), naldxic acid (8.7%) and amikacin (13%).ConclusionsGiardia lamblia, E. Hystolytica/dispar, and Shigella spp are the common etiologies of diarrhea in children in the studied refugee camps. The study also showed that significant numbers of bacterial isolates were resistant to the commonly used antimicrobial drugs. Therefore, improving clinical laboratory services and promoting evidence-based drug prescription may reinforce proper use of antibiotics and reduce the emergence of microbial resistance.

A prevalence survey of enteral parasites in preschool children in the Mangochi District of Malawi

BackgroundHelminthic and protozoan infections are common, particularly in low- or middle-income countries. Although an association between parasite carriage and markers of poor growth have been shown in some studies, systematic reviews have suggested only a modest impact of clearing carriage. The prevalence of these pathogens and the effect that they have on growth in preschool children has never been investigated in Malawi.MethodsOne hundred ninety-three children aged 0–72 months were randomly recruited from rural villages in the Mangochi district of Malawi. Formol-ether concentration was performed on stool and the samples examined with a light microscope. Anthropometric data was taken for each child and the haemoglobin measured with a point of care test.ResultsThe mean age of the children was 2 years 4 months. Overall prevalence of intestinal parasite infection was 37.3%. Protozoa were found in 28.5% of children, while helminths were found in 8.8%. The most commonly found organisms were Giardia lambia (12.4%), Entamoeba coli (10.4%) and Hookworm species (3.6%). Stunting was seen in 47.8% of children, 12.9% were underweight and 5.0% were wasted. No significant association was found between markers of poor growth and infection with any intestinal parasite.ConclusionsWe found that prevalence of helminth infection was low in preschool children living in the Mangochi district compared to international standards. However a significant proportion of the preschool population are infected with protozoa, particularly Giardia lambia. In this cohort, despite a significant prevalence of stunting, helminth infection was not significantly associated with any markers of poor growth. The significance of protozoal carriage and contribution to growth restriction in this context creates further avenues for future research.

1660 Malakoplakia of the Rectum Masquerading as a Tumor: A Case Report

INTRODUCTION: Malakoplakia is a rare inflammatory disorder that occurs in the clinical setting of persistent infections, immunosuppression and tumors. The most common sites of involvement are the urinary, gastrointestinal and respiratory tracts, skin, brain and lymph nodes where they can mimic malignancies. CASE DESCRIPTION/METHODS: A 60-year-old woman with myasthenia gravis and stage IV metastatic thymoma (status-post resection and chemo-radiation) presented with persistent weight loss, bloating, nausea, tenesmus and hematochezia. Her recent history included treatment-resistant Giardia lambia infection for one year as well as recently diagnosed Norovirus and Enteroaggregative Escherichia coli, diagnosed by stool pathogen PCR testing. On colonoscopy, the rectum contained a 7 cm region of polypoid, non-circumferential, non-obstructing nodules (Figure 1) which were highly suspicious for tumor. Microscopically, biopsies revealed rectal lamina propria infiltration by epithelioid cells with small central nuclei, eosinophilic cytoplasm (Figure 2) and lamellated cytoplasmic calcifications (Figure 3). The cells expressed cytoplasmic CD68 (Figure 2) and Von Kossa stain highlighted the calcifications, consistent with Michaelis-Gutmann bodies (Figure 3). Immunostain for pancytokeratin and Ziehl-Neelsen and Gomori's methenamine silver stains were negative. The final diagnosis was malakoplakia. A re-biopsy of the lesion showed positivity for Cytomegalovirus (CMV) immunostain and high levels of CMV viremia (1010IU/mL), for which she was treated with Gancyclovir. DISCUSSION: Malakoplakia is a rare gastrointestinal disorder without specific symptoms, endoscopic features or gross findings and tends to masquerade as a tumor. As illustrated herein, it is often an unexpected diagnosis that depends on pathological recognition of characteristic microscopic features. The correct diagnosis is critical to avoid inappropriate radical treatment. Various micro-organisms have been implicated in its development including bacteria, most commonly E. coli, viruses, fungi and even protozoa such as Taeniaspecies and it is characterized by defective bactericidal activity of the macrophages. Pathologists should be aware that malakoplakia can coexist with malignancy and tuberculosis, therefore, thorough sampling of mass lesions and appropriate adjunctive stains are advisable. Risk factors in our case included an immunocompromised state, multiple concurrent infections, and advanced malignancy, although no malignancy was identified in the biopsies.

Prevalence of Parasites of Public Health Significance in Vegetables Sold in Jos Metropolis, Plateau State, Nigeria

BACKGROUND: Vegetables are vital for good health but can serve as a means of transmission of parasitic pathogens to man. JUSTIFICATION: There is increase consumption of vegetables due to general awareness of the health benefits. This research intends to explore suggestive management strategies and control in handling vegetables to reduce or if possible eliminate food borne parasites. AIM AND OBJECTIVE: The study was to determine parasitic contamination of fresh vegetables with objective to determine the prevalence of parasites of medical importance in vegetables sold in Jos market. METHODOLOGY: A total of 575 samples of fresh vegetables; Lettuce, Cabbage, Carrot, Spinach, Green Pepper, Cucumber, Beetroot, Tomatoes, Garden egg, and Green bean; were bought from sellers from May-June 2016 and examined for helminthes eggs, larva and cysts using wet mount and concentration techniques. RESULT: Hookworm, Ascaris lumbricoides, Trichuris trichuria, Strongyloides stercoralis, cyst of Entamoeba histolytica and Giardia lambia accounted for 7.3%, 8.1%, 4.8%, 58.1%, 9.7% and 12.1% respectively. Cucumber, carrot, cabbage beetroot, spinach, lettuce, green pepper, green beans, garden egg and tomatoes had 0%, 26.7%, 31.7%, 5.7%, 40%, 53.3%, 6.7%, 6.7%, 13.3% and 25% respectively with overall prevalence of 21.6% and strongyloides stercoralis was most prevalent and Trichuris trichiura least prevalent found in cabbage and tomatoes. Lettuce has the highest contamination rate 32(53.3%), followed by spinach 24(40%) while beetroot had the least (5.7%). CONCLUSION: Vegetables in Jos Metropolis are contaminated with parasites. There should be proper washing as they could serve as source of transmission of parasites when eaten raw or undercooked.

Effects of intestinal parasitic infections on nutritional status of primary children in Imo State Nigeria.

IntroductionA cross-sectional study was conducted between the months of April to October 2015, to determine the effects of intestinal parasitic infections (IPIs) on nutritional status of school age children in Owerri and Orlu geographical zones, in Imo State, Nigeria.MethodsFaecal samples were examined using Kato Katz method and formol-ether concentration techniques, while blood samples were examined using cyamethahaemoglobin method. Anthropometric indices were used as indicators of nutritional status, children whose Height-for-Age, Weight-for-Age and Weight-for-Height were <-2 standard deviation (SD) were classified as stunted, wasted, and underweight respectively.ResultsTotal prevalence rate of 16.6% was recorded in the study areas with Ascaris lumbricoides (4.0%), Trichuris trichiura (0.6%), Hookworm (1.0%) Taenia sp (0.3%), Entaomeba histolytica (5.3%), Entamoeba coli (2.7%) and Giardia lambia (2.7) Majority (73.4%) of the children had light intensity. Anthropometric study results showed that 79(31.3%) of the children were malnourished. The prevalence of stunting, under-weight and wasting were higher in uninfected (86.1%, 90.0% and 10%) respectively than in infected children (13.9%, 10.0% and 0.0%) respectively, although not significant at p = 0.857, 0.587 and 0.368 respectively. Prevalence of anaemia was 17.4%, anaemia was insignificantly (p = 0.09) higher in infected (21.1%) than in uninfected (16.5%) children. Children that had co-infection recorded higher prevalence (2.2%) of severe anaemia. There was an association (p = 0.002) between anaemia and intensity of helminth infection. Malnutrition was insignificantly (p = 0.319) higher in children with heavy (100.0%) and moderate (75.0%) intensity of helminth infection than children that had light intensity (41.7%) of helminth infection.ConclusionWhen compared with previous study, there were decline in the prevalence of intestinal parasitic infections and anaemia among school age children. Low intensity parasitemia with intestinal parasites had no significant effect on the malnutrition and haemoglobin profile of the children in the study areas. Therefore, improved sanitation and more deworming efforts should be intensified to ensure further decline in prevalence of intestinal parasitic infections.

Parasitosis Intestinal En Una Población De 5 A 14 Años Que Acuden A Unidades Educativas Escuelas Colegios Públicos De La Ciudad De Riobamba

The purpose of the current research was to determine the prevalence and risk factors linked to intestinal parasites in a population aged from 5 to 14 years who attend to public schools and high schools from Riobamba city, Riobamba-Chambo district. The prospection was carried out between October and November 2015. The population studied was identified and classified according to their age, gender, and educational institution they belong to. In addition, the relation of risk factors linked to the identified parasites was estimated. For this, it was necessary to apply a coprology examination to 2020 boys, girls and teenagers. The fecal samples collected were analyzed using the fresco technique with lugol and saline solution at 0.85%. The total intestine parasite prevalence found was 35.1%. The highest parasite prevalence was detected in groups aged 5 (12.84%), in groups aged 8 (11.71%) and in groups aged 11 (11.56%), while Combatientes de Tapi Educational Unit registered a 34% in both genders. Any significant parasite difference was found among p&lt;0.05 ages. Nine parasite types were identified with Entamoeba coli prevalence (26%) Entamoeba histolytic (9.3%), Iodamoeba bütschlii (0.8%), Giardia lambia (4.3%), Chilomastix mesnilii (0.9%), Hymenolepis nana (1.4%), Ascaris lumbricoides (0.3%) Trichuris trichiura (0.5%), Endolimax nana (1.6%), from which protozoan were the predominant, some poly parasite samples were also found, it was determined that the sample studied reflects a high prevalence of intestinal parasites.

Prevalencia De Parasitosis Intestinales Y Su Posible Relaciòn Con Estados Anèmicos En Los Niños Que Acuden A Los Centros De Educaciòn Inicial

The purpose of this current research is to determine the parasites which has the highest prevalence in the children population in regards to anemia problems in boys and girls aged from 3 to 5 years. The study sample was chosen at random. It was represented by 600 boys and girls from the initial education centers of Riobamba-Chambo district located in Ecuador. From these children, it was possible to obtain 600 stool samples and 600 blood samples that were processed through the fresco technique with lugol and saline solution at 0.85% as well as an automated hematic biometry. The result shows that the total population with parasites was 44,7%. Boys represent the highest parasite prevalence with 51,5% related to the girls with 48,5%. The most frequent parasites were: Entamoeba coli (53,18%), Giardia Lambia (18,15%), Histolytic Entamoeba (14,01%), Endolimax nana (5,42%), lodamoeba butschlii (3,50%), Chilomastix mesnili (3,18%), Hymenolepis nana (1,91), and Ascaris lumbricoides (0,65%). It was concluded that there are no significant relationship between the anemic states and the intestinal parasites on boys and girls since the percentage was 4,5%. It was suggested to the principal of the Riobamba - Chambo District of the Ministry of Education to carry out promotion, prevention, and health care campaigns directed towards the authorities and teachers of the Initial Education Centers as well as parents and their children in order to improve the hygiene and food safety habits of students.

Prevalence of Intestinal Parasites in Vegetables Sold in Some Local Markets in Port-Harcourt, Rivers-State, Nigeria

A total of 492 vegetable leaves samples made up of cabbage (27), bitter leaves (50), garden egg leaves (44), green leaves (66), green onion (47), lettuce (51), pumpkin leaves (111), parsley (26) and water leaves (70), bought randomly from various local markets in Port-Harcourt was analyzed for the presence of some parasite stages. About 145(29.5%) out 492 samples examined were contaminated with various parasite stages. Green leaves had the highest level of contamination (40.9%), followed by bitter leaves (38.8%), while cabbage recorded the lowest (7.4%). Out of 202 parasite stages identified, the eggs of Ascaris lumbricoides were the most predominant 39 (19.3%), followed by the cysts of Entarmoeba histolytica 31 (15.3%), whereas the oocysts Cryptosporidium parvum and eggs of Strongyloides stercolaris had the least 4(1.9%). Green leaves and pumpkin leaves harboured 8 (80.0%) and 7(70%) respectively out of 10 different types of parasites obtained in this study; cabbage and parsley had only 3 (30.0%). Pumpkin leaves recorded the highest level eggs of Ascaris lumbricoides 18(30%), eggs of Fasciola species 10 (16.7%), cysts of Entamaeba histolytica 8 (50.0%), eggs of Giardia lambia 7(11.7%), whereas cabbage overall had the lowest percentage of parasites with Ancylostoma duodenale 2 (50.0%); eggs of Toxoplasma gondii 1 (25.0%) and eggs of Strongyloides stercolaris 1 (25.0%). The present study revealed the potential hazard of vegetable leaves sold in some local markets in Port – Harcourt. Though the vegetables are properly de-contaminated after sales at the point of preparation/cooking, it is also very necessary for the de-contamination to be done before sales. There is need for monitoring by educating both vendors and consumers on the good sanitary practices during processing displaying and sale of the vegetable products and possible danger of contaminated and spoiled vegetables.

Etiological Spectrum of Acute Gastroenteritis in Children Less Than 5 Years of Age with Special Reference to Norovirus

To clarify the infectious etiology of acute gastroenteritis in children less than 5 years of age, we conducted a 1-year prospective case-control study. Stools from150 children with diarrhea and 150 age-matched controls admitted in the hospital for illness involving system other than that of gastrointestinal tract were examined, and their parents were interviewed regarding concerning symptoms. Samples were investigated for parasitic enteropathogens using microscopic examination and bacterial enteropathogens using conventional culture techniques. ELISA was performed for Rotavirus, adenovirus, astrovirus and norovirus were detected by PCR. Overall, a potential pathogen was found in 56.6% of the cases and 4% of controls.. The results indicated that noroviruses (4%) were less common than rotaviruses(24.6%), EPEC (4.6%), and V.Cholerae(4%) but more frequent than astroviruses (2.6%), adenoviruses(3.3%), Giardia lambia(2%) and Shigella spp.(1.3%). In our study it was found that acute gastroenteritis in children less than 5 years is due to viral etiology, where Rotavirus was predominant cause which points towards the need for a vaccine for this pathogen . While EPEC, Vibrio cholerae and Shigella represent the bacterial etiology of pediatric acute gastroenteritis. The role of norovirus as a emerging pathogen responsible for acute gastroenteritis in children needs to be assesed further.

Histomorphometric study of changes in duodenal mucosa of patients presenting with Giardiasis

Context: Giardia lambia is a common intestinal protozoon with a higher prevalence rate in tropical countries as compared to the Western world. Histological changes occur in duodenal mucosa in cases of chronic giardiasis that may be similar to celiac disease or tropical sprue. Aim: As the prevalence of giardiasis is high in our country, the present study was undertaken to study the changes in duodenal mucosa of patients with asymptomatic giardiasis and endoscopically normal duodenum. Study Design: The study included 47 duodenal biopsies out of 372 total cases of endoscopic biopsies examined, in which trophozoites of Giardia were identified. A detailed histo-morphological study of these cases was carried out and the histological findings were evaluated. The major histopathological features as observed in cases of giardiasisi.e inflammation, increased intra epithelial lymphocytes (IELs) and villous abnormalities were compared between patients without giardiasis and those with giardiasis. Statistical Analysis Used: P-value using chi-square test was calculated. Results: P-value was found to be significant for inflammation and villous abnormalities whereas increased IEL's were found to be an insignificant histological parameter. Conclusion: Although the histological features associated with giadiasis were non-specific, a higher incidence of villous changes with increased IEL's was found. This study highlighted the importance of identification of Giardia trophozoites in duodenal biopsies to avoid misdiagnosis of celiac disease or tropical sprue which may have similar clinical and histological features but have different management strategies.

Prevalence of intestinal parasitic infection of kindergarten children

INTRODUCTION: Intestinal parasitic infections are the major public health problems of Nepal. Apart from causing mortality and morbidity, infection with intestinal parasites has been associated with stunting of linear growth, physical weakness and low educational achievement in school children. The drinking water is considered as the major cause for parasitic infection. This study aims to determine the prevalence of parasitic infection among the children visiting kindergarten of the Kathmandu, Khusibhu area. MATERIALS AND METHODS: Cross sectional study was done in randomly selected Kindergarten of Khusibhu area. A total of 101 samples were examined. The study was focused in isolating intestinal parasites using normal saline and iodine wet mount method. Both macroscopic and microscopic studies were performed. RESULTS: Among the samples 45.5% (n=46) showed parasitic infection in which the Giardia lambia infection was found the highest 56.5% (n=26) and infection with Ascaris lumbricoides was found lowest 8.7% (n=4). Infection rate was found to be high in children drinking filtered water than among those drinking boiled and filtered water. CONCLUSION: The prevalence of parasitic infection in children is mainly associated with hygiene and food habit of the children. Hence good education about the hygienic practice helps in the prevention of the infection. DOI: http://dx.doi.org/10.3126/ijim.v2i3.8205 Int J Infect Microbiol 2013;2(3):111-113

The ATP or the natural history of neurotransmission

ATP has become a personal obsession. I am arriving at this exciting field after working in exocytosis for decades. ATP is a natural component of chromaffin granules, the secretory organelle of adrenal chromaffin cells. These cells release catecholamines to the bloodstream as a response to stress stimuli [1]. Natural adrenal catecholamines comprise adrenaline and noradrenaline stored at amazingly high concentrations (0.5–1 M). These huge amounts together with the other soluble components (chromogranins, calcium, ascorbate, protons and ATP) should theoretically promote osmotic forces of about 1,500 mOsm! [2, 7]. To avoid the osmotic lysis of secretory vesicles, the solutes must be aggregated to reduce the tonicity; the main candidates to promote the aggregation have been chromogranins, the most abundant soluble proteins present in chromaffin granules and large dense core vesicles. In an effort to demonstrate this vesicular role of chromogranins, we have studied the exocytotic responses in chromaffin cells from mice lacking in these proteins. Although the absence of chromogranins halves the vesicular amine content and impairs the ability of these secretory vesicles to uptake newly formed catecholamines, the concentration of catecholamines is still very high [5]. This fact made me recall an old paper performed by Ed Westhead's group (University of Massachusetts) in the mid-1980s. Using just an osmometer, they analyzed the osmotic properties of ATP/catecholamine mixtures reporting that this combination behaves as a non-ideal solute resulting in a reduction in the osmolaritay [6]. Recently, Dr. Westhead visited my laboratory for reanalyzing those results on the light of recent data. ATP is co-stored in chromaffin granules with catecholamines at proportion that ranges 1:2.5–6. With these numbers in mind, we obtained the data shown in Fig. 1. After reading the story of co-transmission [3], I guessed that ATP is not co-released with catecholamines; it is just the other way round: amines, peptides, amino acids and practically all of the secreted substances are co-released with ATP, who arrived first (Table 1). Fig. 1 ATP behaves as a non-ideal solute for creating osmotic forces. Plots show the osmolarit0y exhibited by Na–ATP and a mixture of ATP: norepinephrine at a proportion 1/2.8. These experiments were conducted by Dr. Edward Westhead using a vapour-phase ... Table 1 Presence of ATP in secretory vesicles As far as I know, all living eukaryotes have vesicles that accumulate ATP. This includes cells as primitive as Giardia lambia, Trychomonas vaginalis, Toxoplasma gondii or Leishmania donovani, microbes involved in several human parasite diseases [4]. In particular, Giardia is a very intriguing microorganism in that it lacks mitochondria and Golgi apparatus but still has dense granules that contain ATP. One can think that these acidic structures were created as a functional ATP reservoir to preserve it from cytosolic degradation in a cell that may not be efficient enough to produce ATP at a high rate. The conjunction of ATP-containing vesicles and constitutive exocytosis was probably the origin of (neuro)secretion in the days when life started. Later simple biochemical modifications on choline, histidine, tryptophan or tyrosine led to cholinergic, histaminergic, serotonergic and sympathetic transmission. ATP being probably the first neurotransmitter, it seems plausible that the history of secretion started from the fusion of an ATP-contained vesicle with the cell membrane. Newly created transmitters found purinergic vesicles already made. All vesicles accumulate H+: the pH, the electric gradient or both are required for concentrating most transmitters inside. The proton gradient is produced by the V-ATPase, which is also activated by ATP, another fact to think about. Nevertheless, although this commentary will not change any biological fact, we should consider that perhaps exocytosis of ATP is the pillar for understanding cell-to-cell communication.

Characterization and separation of Cryptosporidium and Giardia cells using on-chip dielectrophoresis

Dielectrophoresis (DEP) has been shown to have significant potential for the characterization of cells and could become an efficient tool for rapid identification and assessment of microorganisms. The present work is focused on the trapping, characterization, and separation of two species of Cryptosporidium (C. parvum and C. muris) and Giardia lambia (G. lambia) using a microfluidic experimental setup. Cryptosporidium oocysts, which are 2-4 μm in size and nearly spherical in shape, are used for the preliminary stage of prototype development and testing. G. lambia cysts are 8-12 μm in size. In order to facilitate effective trapping, simulations were performed to study the effects of buffer conductivity and applied voltage on the flow and cell transport inside the DEP chip. Microscopic experiments were performed using the fabricated device and the real part of Clausius-Mossotti factor of the cells was estimated from critical voltages for particle trapping at the electrodes under steady fluid flow. The dielectric properties of the cell compartments (cytoplasm and membrane) were calculated based on a single shell model of the cells. The separation of C. muris and G. lambia is achieved successfully at a frequency of 10 MHz and a voltage of 3 Vpp (peak to peak voltage).

Gene regulation in Giardia lambia involves a putative microRNA derived from a small nucleolar RNA.

Two core microRNA (miRNA) pathway proteins, Dicer and Argonaute, are found in Giardia lamblia, a deeply branching parasitic protozoan. There are, however, no apparent homologues of Drosha or Exportin5 in the genome. Here, we report a 26 nucleotide (nt) RNA derived from a 106 nt Box C/D snoRNA, GlsR2. This small RNA, designated miR5, localizes to the 3′ end of GlsR2 and has a 75 nt hairpin precursor. GlsR2 is processed by the Dicer from Giardia (GlDcr) and generated miR5. Immunoprecipitation of the Argonaute from Giardia (GlAgo) brought down miR5. When a Renilla Luciferase transcript with a 26 nt miR5 antisense sequence at the 3′-untranslated region (3′ UTR) was introduced into Giardia trophozoites, Luciferase expression was reduced ∼25% when synthetic miR5 was also introduced. The Luciferase mRNA level remained, however, unchanged, suggesting translation repression by miR5. This inhibition was fully reversed by introducing also a 2′-O-methylated antisense inhibitor of miR5, suggesting that miR5 acts by interacting specifically with the antisense sequence in the mRNA. A partial antisense knock down of GlDcr or GlAgo in Giardia indicated that the former is needed for miR5 biogenesis whereas the latter is required for miR5-mediated translational repression. Potential targets for miR5 with canonical seed sequences were predicted bioinformatically near the stop codon of Giardia mRNAs. Four out of the 21 most likely targets were tested in the Luciferase reporter assay. miR5 was found to inhibit Luciferase expression (∼20%) of transcripts carrying these potential target sites, indicating that snoRNA-derived miRNA can regulate the expression of multiple genes in Giardia.

Surface-Enhanced Resonance Raman Spectroscopy for the Rapid Detection of Cryptosporidium parvum and Giardia lamblia

A rapid surface-enhanced resonance Raman spectroscopy (SERRS) method has been developed for the detection of two waterborne pathogens, Cryptosporidium parvum and Giardia lambia. Raman labels were prepared by conjugating gold nanoparticles with commercial antibodies and dye molecules. After incubation with the immunogold labels, C. parvum oocysts and G. lamblia could easily be measured and differentiated by Raman spectroscopy. The immunogold signal intensities were optimized by testing several sizes of gold nanoparticles, four different commercially available dye molecules, and two Raman excitation wavelengths. Raman maps were collected across fixed and labeled Cryptosporidium oocysts and Giardia cysts, and the maps were used to determine which C. parvum and G. lamblia antibodies exhibited the best specificities and organism coverages. Ultimately, 40 nm gold nanoparticles were conjugated with rhodamine B isothiocyanate and malachite green isothiocyanate for the C. parvum and G. lamblia immunogold syntheses, respectively. C. parvum monoclonal IgM antibodies and G. lamblia monoclonal IgG1 antibodies resulted in the best immunogold coverage. The research presented here demonstrates the feasibility of utilizing SERRS labeling for sensitive multipathogen monitoring strategies.