Assessing the prevalence and risk factors associated with Entamoeba complex infection among the Orang Asli school children in Perak, Malaysia through molecular approach.

This study performed a cross-sectional investigation on the prevalence of Entamoeba complex infection comprising Entamoeba histolytica, Entamoeba dispar, and Entamoeba moshkovskii and their associated risk factors among the Orang Asli school children in three districts in Perak, Malaysia. Stool samples collected from 544 school children aged between 7 and 12 years old were examined through the nested multiplex PCR assay. The univariate and multivariate regression analyses were then carried out to determine the risk factor associated with Entamoeba complex infection. The overall prevalence of Entamoeba complex infections (E. histolytica, E. dispar and E. moshkovskii) was 21.3% (116/544). Most positive school children were infected with E. moshkovskii (10.7%; 58/544), followed by E. dispar (9.0%; 49/544) and E. histolytica (5.0%; 27/544). Not washing their hands after using the toilet was identified as the only significant risk factor for E. histolytica. The significant risk factors associated with E. moshkovskii infection included children within the age of 10-12 years old, with high BMI, living with working and non-educated mothers, no toilet in the house, not washing their hands after using the toilet, and fever. On the other hand, drinking water from the river, well, and rain was associated with a decreased risk of E. dispar infection. In conclusion, this study showed a high prevalence of Entamoeba spp. infections among the Orang Asli school children in Perak, Malaysia. Addressing the identified risk factors coupled with a holistic approach in breaking the transmission of Entamoeba complex can help improve their quality of life.

Identification and Semi-quantification of Protozoa from the Digestive System Microbiota of the Lobster Cockroach Nauphoeta cinerea Oliver, 1789 (Insecta:Blattaria).

INTRODUCTION: The lobster cockroach Nauphoeta cinerea (N. cinerea) is indicated as a promising non-mammalian model, because it presents behavioral and biochemical alterations also observed in conventional models. In this research, we identified and characterized the distribution of protozoa that inhabit the digestive system (DS) of N. cinerea cockroaches. METHODS: The adult specimens of N. cinerea used in this study (n = 32) were obtained at the Federal University of Santa Maria, dissected and had their visceral contents observed in bright-field microscopy without staining and after application of lugol, Ziehl-Neelsen staining, EA36 trichrome and simulated dark-field microscopy with application of nankin ink. The presence of protozoa in different portions of the DS was semi-quantified by a system of crosses (+). RESULTS: The main taxa observed were: amoebas (Archaemebae:Entamoebida), gregarins (Apicomplexa:Eugregarinide), coccidia (Apicomplexa:Eucoccidiorida), kinetoplastids (Kinetoplastea:Kinetoplastida) and oxymonads (Preaxostyla:Oxymonadida). The highest prevalence of amoebas and gregarines was observed in the medial portion of the DS, while for the other groups, this was seen in the final portion, and in the case of coccidia, such prevalence was specially evidenced by the alcohol-acid coloration. In the present work, the great biological diversity that exists in the microbiota of the digestive system of Nauphoeta cinerea was demonstrated, being possible to find several pathogenic species for humans such as Entamoeba histolytica/dispar/moshkovskii, Cryptosporidium sp. and Cyclospora cayetanensis. There is still a lot to know about the interactions between endocommensal protozoa and their respective invertebrate hosts, so the best way to clarify such relationships is through molecular and genetic test.

Detection of enteropathogens and research of pesticide residues in Lactuca sativa from traditional and agroecological fairs / Detecção de enteropatógenos e pesquisa de resíduos de agrotóxicos em Lactuca sativa de feiras tradicionais e agroecológicas

Agroecological production represents a new reality in vegetable farming. Thus, the study aimed to search for the presence of pesticides in agroecological samples and microbiological and parasitological contamination in Lactuca sativa L. of agroecological and conventional fairs in Alagoas. Thirty-two samples were collected, a hygienicsanitary checklist was performed, along with coliform, Salmonella spp., Staphylococcus aureus research and Gas Chromatography with Mass Spectrometer (GC-MS) for pesticide research; using two methodologies for parasitological studies. Thirty-two samples were analyzed, with a sanitary adequacy level at 45% and 38% for fairs (agroecological and conventional) respectively, with three samples (9.4%) in total, being adequate to the microbiological pattern according to RDC no. 12/2001. Among the 23 samples which were tested positive for parasites, 69.5% originated from samples extracted at conventional fairs, and 53% of every positive samples presented Entamoeba coli as the main contaminant. All together, no traces of deltamethrin were detected, but in two of the samples a peak retention for diphenoconazole was detected. In conclusion, the fairs represent an adequate environment for human pathogens and indicators. Therefore greater hygiene training of vegetable traders and consumers in the state of Alagoas is required, especially in conventional fairs, which presented more critical microbiological and parasitological indexes comparing to fairs with agroecological products; however, the presence of difenoconazole in two samples of agricultural fairs suggests accidental contamination or indiscriminate use of this chemical in the production of Lactuca sativa, however, products with agroecological origin represents a better choice for the consumers.

A produção agroecológica representa uma nova realidade no cultivo das hortaliças. Assim, o estudo teve como objetivo verificar a presença de agrotóxicos em amostras agroecológicas e analisar a contaminação microbiológico e parasitológico em Lactuca sativa L. de feiras agroecológicas e convencionais de Alagoas. Foram coletadas 32 amostras, realizando checklist higiênico-sanitário, análises de coliformes, Salmonella spp., Staphylococcus aureus e Cromatografia Gasosa com Espectrômetro de Massas (GC-MS) para pesquisa de agrotóxicos; utilizando-se duas metodologias para estudos parasitológicos. Foram analisadas 32 amostras, com nível de adequação sanitária de 45% e 38% para feiras (agroecológicas e convencionais) respectivamente, sendo três amostras no total (9,4%) adequadas a padrão microbiológico segundo RDC n° 12//2001; Dentre as 23 amostras positivas para parasitos, 69,5% teve origem em amostras de feiras convencionais, e 53% de todas as amostras positivas apresentaram Entamoeba coli como principal contaminante, juntamente, não foi detectado traços de deltametrina, porem foi visualizado em duas amostras, pico de retenção para padrão de difenoconazol. Concluindo-se que as feiras representam um ambiente adequado para patógenos humanos e indicadores, necessitando de uma maior higienização e treinamento dos comerciantes e consumidores de hortaliças do estado de Alagoas, principalmente em feiras convencionais, que apresentaram índices microbiológicos e parasitológicos mais críticos em relação a feiras com produtos agroecológicos; mas, a presença de difenoconazole em duas amostras de feiras agroecológicas sugerem contaminação acidental ou uso indiscriminado deste químico em produção da Lactuca sativa L., todavia os produtos de origem agroecológica representam uma melhor escolha para o consumidor.

Ocorrência de enteroparasitas em alfaces (Lactuca sativa) comercializadas em mercados municipais em Rio Branco, Acre / Occurrence of enteroparasitas in alfaces (Lactuca sativa) commercialized in municipal markets in Rio Branco, Acre / Presencia de enteroparasitos en lechuga (Lactuca sativa) comercializados en mercados municipales de Rio Branco, Acre

As hortaliças folhosas são ricas em vitaminas, sais minerais e fibras alimentares, proporcionando importantes benefícios para a saúde, o que acaba atraindo os consumidores, que costumam ingeri-las na forma crua, ficando expostos à contaminação por enteroparasitas. O objetivo do trabalho foi verificar a possível presença de enteroparasitas em alfaces (Lactuca sativa) comercializadas em três mercados municipais de Rio Branco, Acre. Foram analisadas 26 amostras de alfaces, onde as folhas das alfaces foram lavadas, em uma solução contendo 250 mL de água destilada e cinco gotas de detergente. O líquido foi filtrado em gazes de oito dobras e deixado sedimentar por 24 horas, à temperatura ambiente. Após o tempo decorrido o sobrenadante foi descartado, foram preparadas lâminas, que foram analisadas ao microscópio óptico. Os resultados obtidos neste estudo evidenciaram contaminação em 96,2% do total de amostras de alfaces analisadas. As principais estruturas parasitárias encontradas foram: Ascaridia sp., Eimeria sp., Entamoeba coli, Entamoeba histolytica, Ancylostoma sp., Strongyloides stercoralis, Enterobius vermicularis, Dipylidium caninum, Trichuris trichiura, Diphillobothrium latum e Balantidium sp. O enteroparasita mais abundante foi Ascaridia sp., tendo sido encontrados 164 (430) exemplares nas amostras, representando 38,14% do total de enteroparasitas encontrados. Ascaridia sp. foi também o enteroparasita mais frequente, sendo encontrada em 42,3% do total de amostras analisadas. Nesse contexto, a ingestão de alfaces cruas, provenientes dos mercados municipais estudados, pode representar risco potencial para os consumidores, caso elas não sejam higienizadas corretamente. Isto demonstra a importância de uma boa limpeza e desinfecção das folhas de alface antes do consumo.(AU)

Leafy vegetalizes are reich in vitaminas, mineral and. deitar febres, providing important health benefits, which ends up attracting consumers, who usually eat them in the raw form, being exposed to contamination by enteroparasites. The objective of the study was to verify the possible presence of enteroparasites in lettuce (Lactuca sativa) sold in three municipal markets in Rio Branco, Acre. 26 samples of lettuce were analyzed, where the lettuce leaves were washed, in a solution containing 250 mL of distilled water and. five drops of detergent. The liquid was filtered through eight-fold gauze and. left to settle for 24 hours at room temperature. After the elapsed time, the supernatant was discarded, slides were prepared and. analyzed under an optical microscope. The results obtained in this study showed contamination in 96.2% of the total samples of lettuce analyzed. The main parasitic structures found were: Ascaridia sp., Eimeria sp., Entamoeba coli, Entamoeba histolytica, Ancylostoma sp., Strongyloides stercoralis, Enterobius vermicularis, Dipylidium caninum, Trichuris trichiura, Diphillobothrium latum e Balantidium sp. The most abundant enteroparasite was Ascaridia sp., 164 (430) specimens were found in the samples, representing 38.14% of the total enteroparasites found. Ascaridia sp. was also the most common enteroparasite, was found in 42.3% of the total samples analyzed. In this context, ingestion of raw lettuce from the municipal markets studied may pose a potential risk to consumers if they are not properly sanitized. This demonstrates the importance of good cleaning and disinfection of lettuce leaves before consumption.(AU)

Las verduras de hoja son ricas en vitaminas, minerales y fibras dietéticas, aportando importantes beneficios para la salud, lo que acaba atrayendo a los consumidores, que suelen consumirlas crudas, quedando expuestas a la contaminación por enteroparásitos. El objetivo del estudio fue verificar la posible presencia de enteroparásitos en lechugas (Lactuca sativa) vendidas en tres mercados municipales de Rio Branco, Acre. Se analizaron 26 muestras de lechuga, donde se lavaron las hojas de lechuga, en una solución que contenía 250 mL de agua destilada y cinco gotas de detergente. El líquido se filtró a través de una gasa de ocho veces y se dejó reposar durante 24 horas a temperatura ambiente. Transcurrido el tiempo, se descartó el sobrenadante, se prepararon los portaobjetos y se analizaron al microscopio óptico. Los resultados obtenidos en este estudio mostraron contaminación en el 96,2% del total de muestras de lechuga analizadas. Las principales estructuras parasitarias encontradas fueron: Ascaridia sp., Eimeria sp., Entamoeba coli, Entamoeba histolytica, Ancylostoma sp., Strongyloides stercoralis, Enterobius vermicularis, Dipylidium caninum, Trichuris trichiura, Diphillobothrium latum y Balantidium sp. El enteroparásito más abundante fue Ascaridia sp., Se encontraron 164 (430) ejemplares en las muestras, lo que representa el 38,14% del total de enteroparásitos encontrados. Ascaridia sp. también fue el enteroparásito más frecuente, encontrándose en el 42,3% del total de muestras analizadas. En este contexto, la ingesta de lechuga cruda, de los mercados municipales estudiados, puede representar un riesgo potencial para los consumidores, si no se higienizan adecuadamente. Esto demuestra la importancia de una buena limpieza y desinfección de las hojas de lechuga antes del consumo.(AU)

Hypericum erectum alcoholic extract inhibits Toxoplasma growth and Entamoeba encystation: an exploratory study on the anti-protozoan potential.

Hypericum erectum is an important ethnobotanical medicine in East Asian tradition. To explore the anti-parasitic potential of H. erectum, inhibitory effects on the growth of intracellular parasite Toxoplasma and on the encystation of intestinal parasite Entamoeba were examined. The constituents in H. erectum alcoholic extracts and fractions separated by solvent-partitioning were analysed by high resolution LC-MS. Toxoplasma gondii growth inhibition assay was performed using GFP-labelled T. gondii strain PTG-GFP by measuring the fluorescence intensity. Anti-Toxoplasma drug pyrimethamine was used as a positive control. T. gondii-induced immune reaction was assessed by quantitative PCR and fluorescence microscopy, using co-culture of PTG-GFP and monocyte-macrophage cell line Raw264. The inhibitory effect on the encystation of Entamoeba invadens was measured by flow-cytometry, where paromomycin was used as a positive control. H. erectum methanol (MeOH) extract (50 µg/mL) and ethyl acetate (EtOAc) fraction (50 µg/mL) inhibited the growth of T. gondii, while 50%MeOH extract and hydrophilic fractions were ineffective. Co-culture with T. gondii reduced the viability of macrophages, however macrophages were protected in the presence of H. erectum MeOH extract or EtOAc fraction (above 10 µg/mL). The MeOH extract and EtOAc fraction also effectively suppressed the encystation of E. invadens at 1 mg/mL. Hypericine, a major constituent in MeOH extract and EtOAc fraction, inhibited T. gondii growth and E. invadens encystation. Our results demonstrated that H. erectum effectively inhibited Toxoplasma growth and Entamoeba encystation. These activities are partly mediated by hypericin. In addition, it was suggested the extract and fraction may protect innate immune cells from Toxoplasma-induced damages, thereby enhancing parasite clearance. Further investigation is warranted to address the in vivo effectiveness of H. erectum as an anti-protozoal medicine.

High-Throughput Screening of Entamoeba Identifies Compounds Which Target Both Life Cycle Stages and Which Are Effective Against Metronidazole Resistant Parasites.

Neglected tropical diseases, especially those caused by parasites, are significantly underserved by current drug development efforts, mostly due to the high costs and low economic returns. One method for lowering the costs of drug discovery and development for these diseases is to repurpose drugs developed for other indications. Here, we present the results of a screen of five repurposed drug libraries to identify potential new lead compounds to treat amebiasis, a disease that affects tens of millions of people and causes ~100,000 deaths annually. E. histolytica, the causative agent of amebiasis, has two major life cycle stages, the trophozoite and the cyst. The current primary treatment for amebiasis, nitroimidazole compounds, do not eliminate parasites from the colonic lumen, necessitating a multi-drug treatment regimen. We aimed to address this problem by screening against both life stages, with the aim of identifying a single drug that targets both. We successfully identified eleven compounds with activity against both cysts and trophozoites, as well as multiple compounds that killed trophozoites with improved efficacy over existing drugs. Two lead compounds (anisomycin and prodigiosin) were further characterized for activity against metronidazole (MNZ) resistant parasites and mature cysts. Anisomycin and prodigiosin were both able to kill MNZ resistant parasites while prodigiosin and its analog obatoclax were active against mature cysts. This work confirms the feasibility of identifying drugs that target both Entamoeba trophozoites and cysts, and is an important step toward developing improved treatment regimens for Entamoeba infection.

Evaluación de la contaminación microbiológica de la lechuga (lactuca sativa) en la cadena alimentaria, provincia de Quillacollo, Cochabamba, Bolivia 2015 / Evaluation of microbiological contamination of lettuce (Lactuca sativa) in the food chain, province of Quillacollo, Cochabamba, Bolivia 2015

Las enfermedades transmitidas por alimentos constituyen un problema de salud, diversos estudios demuestran la alta contaminación microbiológica de la lechuga en la cadena alimentaria. El objetivo de la presente investigación fue evaluar la contaminación microbio-lógica de la lechuga (Lactuca sativa) en la cadena alimentaria, en la provincia Quillacollo-Cochabamba-Bolivia. Muestras de lechugas fueron tomadas en los meses de abril y mayo, con el mismo de número de muestras, en tres puntos de la cadena alimentaria: parcelas de producción (localidad Combuyo), puestos de venta de lechuga y de comida de los Mercados 9 de diciembre y Central de Quillacollo. El procesamiento de las muestras fue realizado en los laboratorios de Microbiología de LABIMED, y Post Grado de Medicina Tropical de la Facultad de Medicina, UMSS. Los resultados demuestran la contaminación de las lechugas con bacterias Aerobias Mesófilas, colifor-mes totales y fecales en concentraciones superiores a los estándares permitidos en los tres puntos de la cadenas alimentaria evaluadas; identificándose Salmonella sp en una de las muestras de lechugas procesada de los puestos de venta de comida de los mercados. La contaminación parasitaria fue evidenciada en los puestos de venta solo en el mes de mayo en 83% (n=10) de las 12 muestras analizadas, detectándose quistes de Entamoeba coli y larvas de Strongyloides sp, contaminación relacionada con la utilización de abono de aves en el cultivo y la manipulación inadecuada de la lechuga en la cadena alimentaria. En conclusión podemos indicar que las lechugas presentan una importante contaminación microbiana en los tres puntos evaluados de la cadena alimentaria, por lo que constituye un riego para la salud pública.

Infectious diseases transmitted by food, are considered a public health issue, several studies shown the high microbiological contamination of the letucce in particular (Lactuca sativa). The aim of this study was to evaluate the contamination of letucce at Quillacollo Province of Cochabamba- Bolivia. The samples were taken between april and may, with the same number of samples at production plots (Combuyo location), and at selling places of vegetables and food (markets) such as "9 de diciembre” market and "Central Quillacollo” market. The samples were processed at LABIMED laboratories (microbiology, and Tropical Medicine Post grade of medicine Faculty of San Simón University). Results shows contamination of lettuce with Aerobic Mesophilic Bacterias, and total coliforms and fecal coliforms, in higher concentration than standard allowed, at any point of chain food that has been evaluate; Sallmonella sp was identify at selling points with 83% of the samples and only at the month of may (n=10 of 12 samples examined) also Entamoeba coli cysts and Strongyloides sp worms; the contamination was associate with bird fertilizer at crops and at inapropiate food management of the lettuce. Therefore we can conclude that lettuce's contamination is present at the three evaluated points of its chain, production selling points until consume, wich must be considered as a public health risk.

Fitoterapia Mbyá-Guaraní en el control de las parasitosis intestinales. Un estudio exploratorio con Chenopodium Ambrosioides L. Var. Anthelminticum en cinco comunidades de misiones, Argentina / Phytotherapy Mbyá-Guaraní in the control of intestinal parasitoses. An exploratory study with Chenopodium Ambrosioides L. Var. Anthelminticum in five communities in Misiones, Argentina

En las comunidades Mbyá-Guaraní, el conocimiento local de las especies vegetales que habitan el "monte" se pone de manifi esto en el desarrollo de distintas actividades tales como su empleo medicinal en las parasitosis y dolencias asociadas. El objetivo de esta contribución fue evaluar la efi cacia de Chenopodium ambrosioides L. var. anthelminticum (Ka´aré) en el control de las enteroparasitosis. El estudio fue realizado durante septiembre de 1998 y abril de 1999 en las comunidades de Marangatú (MA), Ñamandú (ÑA), El Pocito (PO), Tabay (TB) y Takuapí (TA) del Departamento Libertador General San Martín, Misiones, Argentina. La muestra incluyó 148 personas que fueron agrupadas por sexo e intervalos etáreos. El tratamiento fi toterapéutico consistió en la administración de tintura de Ka´aré (maceración en alcohol etílico al 70%). El análisis coproparasitológico se realizó mediante examen directo y la técnica de enriquecimiento por fl otación (Füllerbon) previo y posterior al tratamiento con el Ka´aré. Los resultados indicaron altos porcentajes de individuos parasitados previo al tratamiento (PRET) que fl uctuaron entre 73.5% y 91.7%. Entre los protozoos Entamoeba coli fue la especie más prevalente con porcentajes entre 18.4% y 37.5%. Hymenolepis nana entre los cestodes presentó prevalencias entre 4.2% y 20.8%. Entre los nemátodos Ancylostoma duodenale/Necator americanus (Ancylostomideos) fue la más frecuente y su prevalencia fl uctuó entre 55.1% y 83.3%. Trichuris trichiura sólo fue observado en ÑA y con una prevalencia muy baja (3.8%). El porcentaje de monoparasitismo fue mayor en ÑA (56.5%), el de biparasitismo en MA y TA (44.4%), mientras que el de poliparasitismo en TB (36.8%). En la evaluación postratamiento (POST) la prevalencia total de parasitosis descendió respecto del pretratamiento (79.7% vs 68.9%) (X2 Yates = 4.57; p < 0.05). Cuando se analizó por sexo, el porcentaje de mujeres parasitadas fue menor, con diferencia estadísticamente signifi cativa (X2 MantelHaenszel = 4.28; p < 0.05). Asimismo, en los individuos mayores de 30 años (intervalo 3) la diferencia fue estadísticamente significativa entre pre y postratamiento (90.9% vs. 50.0%) (X2 Yates = 6.99; p < 0.01). En los individuos postratamiento se observó una disminución en los valores de prevalencia de E. coli (30.8%-0.0%) y Ancylostomídeos (52.0%-11.5%). Así, en ÑA los Ancylostomídeos mostraron diferencias estadísticamente significativas (73.1% vs. 11.5%) (X2 Yates = 8.99; p < 0.05). Asimismo, la prevalencia de Ascaris lumbricoides descendió en los individuos de todas las comunidades, siendo más evidente en PO (20.8% vs. 0.0%) y en ÑA (19.2% vs. 0.0%). Para Giardia lamblia, H. nana y Strongyloides stercoralis el tratamiento fi toterapeútico no fue efectivo. Respecto al monoparasitismo se observó que sólo disminuyó levemente en TA (44.4% vs. 43.2%) y aumentó en el resto de las comunidades, siendo mayor el aumento de monoparasitados en ÑA (56.5% vs. 76.9%). Sin embargo el porcentaje de biparasitismo disminuyó en la mayoría de las comunidades estudiadas, con excepción de TB (36.8% vs. 50.0%). Por último el poliparasitismo descendió sólo en TB (36.8% vs. 11.1%) y ÑA (17.4% vs.7.7%). Los resultados obtenidos demostraron la efectividad de Ch. ambrosioides L. var. anthelminticum sobre los Ancylostomideos y A. lumbricoides, mientras que para protozoos, cestodes y larvas de S. stercoralis no fue efectivo.

Iron-sulphur clusters, their biosynthesis, and biological functions in protozoan parasites.

Fe-S clusters are ensembles of sulphide-linked di-, tri-, and tetra-iron centres of a variety of metalloproteins that play important roles in reduction and oxidation of mitochondrial electron transport, energy metabolism, regulation of gene expression, cell survival, nitrogen fixation, and numerous other metabolic pathways. The Fe-S clusters are assembled by one of four distinct systems: NIF, SUF, ISC, and CIA machineries. The ISC machinery is a house-keeping system conserved widely from prokaryotes to higher eukaryotes, while the other systems are present in a limited range of organisms and play supplementary roles under certain conditions such as stress. Fe-S cluster-containing proteins and the components required for Fe-S cluster biosynthesis are modulated under stress conditions, drug resistance, and developmental stages. It is also known that a defect in Fe-S proteins and Fe-S cluster biogenesis leads to many genetic disorders in humans, which indicates the importance of the systems. In this review, we describe the biological and physiological significance of Fe-S cluster-containing proteins and their biosynthesis in parasitic protozoa including Plasmodium, Trypanosoma, Leishmania, Giardia, Trichomonas, Entamoeba, Cryptosporidium, Blastocystis, and microsporidia. We also discuss the roles of Fe-S cluster biosynthesis in proliferation, differentiation, and stress response in protozoan parasites. The heterogeneity of the systems and the compartmentalization of Fe-S cluster biogenesis in the protozoan parasites likely reflect divergent evolution under highly diverse environmental niches, and influence their parasitic lifestyle and pathogenesis. Finally, both Fe-S cluster-containing proteins and their biosynthetic machinery in protozoan parasites are remarkably different from those in their mammalian hosts. Thus, they represent a rational target for the development of novel chemotherapeutic and prophylactic agents against protozoan infections.

[Diabetic foot infections. Prevalence and antibiotic sensitivity of the causative microorganisms]. / Infecciones del pie diabético. Prevalencia de los distintos microorganismos y sensibilidad a los antimicrobianos.

BACKGROUND: Foot infections are a common reason for hospitalization and a cause of complications in patients with diabetes. The aim of this study was to determine the prevalence of microorganisms found on culture in complicated diabetic foot infections in hospitalized patients, and the sensitivity of the causative microorganisms to antimicrobial agents. METHODS: Between December 2001 and December 2005 in our department, 84 samples in 62 diabetic patients with moderate/severe infection were collected for microbiological study. RESULTS: At least one microorganism was isolated in 88% of samples. The most frequently isolated germ group was gram-positive bacteria (55% of the samples), with Staphylococcus aureus (33%) in the first position, followed by Pseudomonas aeruginosa (12%), Enterococcus spp. (9%), and Escherichia coli (8%). Culture for anaerobic microorganisms was only performed in half the samples; 25% were positive, and Peptostreptococcus spp. predominated. Among the multiresistant microorganisms, methicillin-resistant staphylococci aureus (MRSA) were the most common, accounting for 38% of the isolated strains of S. aureus, ie, 12% of all samples. As to the gram-negative microorganisms, nearly 30% of E. coli strains were resistant to amoxicillin/clavulanic acid and ciprofloxacin. CONCLUSION: Most of the cultures in our study were monomicrobial, with S. aureus being the most prevalent microorganism, followed by enterobacteria and P. aeruginosa. The main resistant microorganism in diabetic foot infections requiring hospitalization was methicillin-resistant golden staphylococcus, which was found in 12% of the series.

Growth-promoting effect on iron-sulfur proteins on axenic cultures of Entamoeba dispar.

A growth-promoting factor (GPF) that promotes the growth of Entamoeba dispar under axenic culture conditions was found in fractions of mitochondria (Mt), hydrogenosomes (Hg) and chloroplasts (Cp) obtained from cells of six different protozoan, mammalian and plant species. We were able to extract the GPF from the Cp-rich leaf cells of a plant (spiderwort: Commelina communis L.) in an acetone-soluble fraction as a complex of chlorophyll with low molecular weight proteins (molecular weight [MW] approximately 4,600). We also found that on treatment with 0.6% complexes of 2-mercapthoethanol (2ME), complexes of chlorophyll-a with iron-sulphur (Fe-S) proteins (e.g., ferredoxins [Fd] from spinach and Clostridium pasteurianum) and noncomplex rubredoxin (Rd) from C. posteurianum have a growth-promoting effect on E. dispar. These findings suggest that E. dispar may lack a sufficient quantity of some essential components of Fe-S proteins, such as Fe-S center.

Entamoeba invadens: in vitro axenic encystation with a serum substitute.

The current media for axenic cultivation of Entamoeba histolytica and Entamoeba invadens are supplemented with bovine or equine serum, which provides several essential nutrients to amoebas. Serum has also been considered an essential component in encystation media for E. invadens. A substitute of serum, PACSR has been described as an alternative for growth of E. histolytica and also maintains growth of E. invadens. When PACSR was used instead of serum for encystation of E. invadens the efficiency was the same as for serum. Our present data show that PACSR can support the growth and induction of encystation of E. invadens strain IP-1.

Axenic cultivation of Entamoeba dispar in newly designed yeast extract-iron-gluconic acid-dihydroxyacetone-serum medium.

Yeast extract-iron-gluconic acid-dihydroxyacetone-serum medium that allows axenic cultivation of Entamoeba dispar was designed based on casein-free yeast extract-iron-serum (YI-S) medium, and the usefulness of the medium was assessed. The main differences from YI-S medium are replacement of glucose by gluconic acid, addition of dihydroxyacetone and D-galacturonic acid monohydrate, and sterilization by filtration. This medium promoted the axenic growth of 5 strains of E. dispar (2 strains of nonhuman primate isolates and 3 strains of human isolates). In addition, to clarify the biological basis for the growth of E. dispar in this medium, analyses of relevant enzymes on the glycolytic pathway of the amoebae as well as of the protozoans that are the best culture supplement for amoebae are being performed.

Freqüência de enteroparasitas em amostras de alface (Lactuca sativa) comercializadas em Lavras, Minas Gerais / Frequency of intestinal parasites in samples of lettuce (Lactuca sativa) commercialized in Lavras, Minas Gerais State

O objetivo deste trabalho foi realizar uma avaliaçäo parasitológica em amostras de alface (Lactuca sativa) comercializadas em Lavras, MG. As amostras de alfaces apresentaram baixos padröes higiênicos, indicados pela presença de formas parasitológicas de origem animal ou humana e alta concentraçäo de coliformes fecais

Contaminaçäo microbiológica e parasitológica em alfaces (Lactuca sativa) de restaurantes self-service, de Niterói, RJ / Microbiological and parasitological contamination of lettuce (Lactuca sativa) from self service restaurants of Niterói city, RJ

Analisou-se 30 amostras de alfaces de restaurantes self-service de Niterói quanto à presença de bactérias e parasitas. Dezesseis amostras apresentaram coliformes fecais, 16 mesófilos acima de 10(7) UFC/g e 3 cistos de Entamoeba coli. Estes dados indicam a necessidade da orientaçäo dos manipuladores quanto a higienizaçäo no preparo de hortaliças

The presence of antiamoebic constituents in psyllium husk.

The crude extract of psyllium husk (ispaghula) and its active constituent (petroleum fraction) caused varying degrees of growth inhibition in three different species of Entamoeba, i.e. Entamoeba histolytica, E. invadens and E. dispar. The inhibitory effect of the crude extract was in the dose range of 1-10 mg/mL, whereas a similar inhibitory effect was obtained with the petroleum fraction at a much lower dose (0.1-1.0 mg/mL), indicating that the active chemical(s) is/are concentrated in the petroleum fraction. These data support the traditional use of psyllium husk in amoebic dysentery.

Alkaloids from Cyrtanthus elatus.

The alcoholic extract of the fresh bulbs of Cyrtanthus elatus yielded zephyranthine (1) and 1,2-O-diacetylzephyranthine (2), together with three other known alkaloids. Complete assignment of 1H and 13C NMR spectra of compounds 1 and 2 was done by employment of two-dimensional NMR techniques.

Antiamoebic activity of Piper longum fruits against Entamoeba histolytica in vitro and in vivo.

The fruits of Piper longum used in traditional remedies against intestinal distress have been tested for their efficacy against experimental caecal amoebiasis of rats. The ethanolic extract, hexane fraction, n-butanol soluble fraction exerted in vitro amoebicidal action at 1000 micrograms/mL and the chloroform fraction at 500 micrograms/mL. The ethanolic extract and piperine, a pure compound, from this plant material cured 90% and 40% of rats with caecal amoebiasis, respectively.

[Growth inhibition of Entamoeba histolytica and E. invadens produced by pomegranate root (Punica granatum L.)]. / Inhibición del crecimiento de Entamoeba histolytica y E. invadens producida por la raíz del granado (Punica granatum L.).

The purpose of the present work has been to evaluate the biological activity of alkaloids and tannins extracted from roots of Punica granatum L. on axenic cultures from Entamoeba histolytica and E. invadens, strains. Initially, the total aqueous extract and later some of its components obtained by thin layer chromatography were tested. These compounds were identified by chromatography as alkaloids or tannins. The density of amebic cultures was determined with a hemocytometer after 48 and 96 hours of incubation, which was calculated by the difference between number of trophozoites obtained at the times chosen and the number of amoebae inoculated. Two milliliters of aqueous extract had higher activity on cultures from E. histolytica than E. invadens strains, producing growth inhibitions of about 100 and 40 per cent respectively. Alkaloid concentrations of 1 mg/ml had no amebicide activity, however tannins at concentrations of 10 micrograms/ml for E. histolytica, and 100 micrograms/ml for E. invadens were sufficient to produce an growth inhibition about 100 per cent. Tannic acid was also tested on the cultures of E. histolytica observing an high inhibitory activity on the growth, this effect was produced at 0.01 mg/ml was similar to that observed with the tannins mixture.