Assessment of dead-end ultrafiltration for the detection and quantification of microbial indicators and pathogens in the drinking water treatment processes.

A safe water supply requires distinct treatments and monitoring to guarantee the absence of pathogens and substances potentially hazardous for human health. In this study we assessed the efficiency of the dead-end ultrafiltration (DEUF) method to concentrate faecal indicator organisms (FIO) and pathogens in water samples with different physicochemical characteristics. Water samples were collected at the treatment stages of two drinking water treatment plants to analyse the concentration of a variety of 7 FIO and 4 reference microbes which have some species that are pathogenic to humans: Campylobacter spp., enteroviruses, Cryptosporidium spp. and Giardia spp. The samples were analysed before and after concentration by DEUF, detecting FIO concentrations about 1 log10 higher in non-concentrated samples from both catchments. Percent recoveries were highly variable with a mean of 43.8 ± 17.5%, depending on the FIO and inherent sample characteristics. However, DEUF enabled FIO concentration in high volumes of water (100-500 l), allowing a reduction in the detection limit compared to the non-concentrated samples due to the high volume processing capabilities of the method. As a consequence, the detection of FIO removal from water in the drinking water treatment process was 1.0-1.5 logarithms greater in DEUF-treated water compared to unfiltered samples. The DEUF method improved the detection of target indicators and allowed for the detection of pathogens in low concentrations in water after the treatment stages, confirming the suitability of DEUF to concentrate high volumes of different types of water. This method could be useful for microbial analysis in water treatment monitoring and risk assessment, allowing the identification of critical points during the water treatment process and potential hazards in water destined for several uses.

Brachylaimiasis: Brachylaima spp. (Digenea: Brachylaimidae) Metacercariae Parasitizing the Edible Snail Cornu aspersum (Helicidae) in Spanish Public Marketplaces and Health-Associated Risk Factors.

The edible land snail Cornu aspersum (Pulmonata: Stylommatophora) acts as a second intermediate host in the terrestrial life cycle of Brachylaima spp. trematodes, harboring unencysted metacercariae in its kidney. The ingestion of undercooked infected snails by humans may allow metacercariae to potentially develop to adult stage in the intestine, causing brachylaimiasis, as already seen in Australia. The prevalence and dynamics of C. aspersum parasitization by Brachylaima spp. metacercariae in specimens intended for human consumption in Spanish marketplaces were studied. In total, 3,710 C. aspersum specimens were analyzed over 5 yr, which were obtained from public marketplaces in the Spanish cities of Barcelona, Bilbao, Madrid, Tudela, Valencia, and Zaragoza. The overall prevalence was 41.97% (95% CI: 40.38-45.56%). The Tudela marketplace had the highest values for both the seasonal prevalence and abundance in all studies during autumn (93.57% and 3.09, respectively). This market also gave the highest individual metacercarial burden recorded, 212 metacercariae in a single specimen. Overall, the highest prevalence of Brachylaima spp. occurred in autumn (58.65%) and the lowest in winter (22.64%). There was a seasonal effect on prevalence, which increased from summer to autumn and then decreased in winter. In total, 96 experimental Brachylaima adults were obtained from the metacercariae parasitizing the analyzed snails. These were identified through morphometric tools (principal component analysis) as Brachylaima mascomai (56 in Barcelona, 1 in Bilbao, 7 in Tudela, and 3 in Valencia), and Brachylaima llobregatensis (17 in Barcelona, 8 in Bilbao, 1 in Valencia, and 3 in Zaragoza). Logistic regression modeling, conducted to predict the probability of purchasing parasitized snails using city and season as predictors showed a correct prediction overall of 79.0%, with a significant (p = 0.001) risk effect in the Barcelona-autumn interaction (2.551-38.442), a significant (p = 0.049) protection effect in the Tudela-spring interaction (0.076-0.997), a significant (p < 0.001) risk effect in the Tudela-autumn interaction (4.330-78.584), and a significant (p = 0.014) protection effect in the Valencia-spring interaction (0.033-0.687). The high overall prevalence of Brachylaima spp. metacercariae should be a matter of concern for public health authorities, mainly in countries where C. aspersum is consumed.

Effect of Praziquantel on the Tegument and Digestive Epithelium Ultrastructure of Brachylaima sp. Metacercariae Parasitizing the Edible Land Snail Cornu aspersum.

The edible land snail Cornu aspersum (Pulmonata: Stylommatophora) acts as second intermediate host in the cycle of Brachylaima sp. trematode, harboring free metacercariae in its kidney. The ingestion of undercooked infected snails by humans allows metacercariae to develop to adult stage in the intestine, causing brachylaimiasis. Praziquantel (PZQ) is the drug of choice to treat trematodiasis and it is effective against Brachylaima sp. metacercariae. The objective of this work was to assess, by transmission electron microscopy (TEM), the ultrastructural changes produced on the tegument and gastrodermis of the Brachylaima metacercariae recovered from C. aspersum treated with PZQ in comparison with untreated ones. Snails naturally infected by Brachylaima sp. metacercariae were treated by PZQ both individually and in groups. Metacercariae recovered from treated and control snails were processed for TEM. The tegument of untreated metacercariae was covered by a regular and thick glycocalyx. The syncytial epithelium contained abundant T2 secretory bodies appearing as membrane-bound biconcave disk-vesicles with high electron-dense and uniform content. The T2 secretory bodies located along the external area of the syncytium were mainly arranged at right angles to the apical plasma membrane. In treated metacercariae, the content of the T2 secretory bodies appeared altered, degenerating from high to low electron density, losing its uniform appearance and forming high electron-dense accumulations scattered around the periphery of the vesicle and separated by low electron-dense spaces. The presence of clusters was detectable in the central area. The characteristic arrangement of the T2 secretory bodies observed in untreated metacercariae was lost in treated ones. Vesicles near the apical area of the tegument no longer maintained their arrangement perpendicular to the apical plasma membrane. The characteristic arrangement of T2 secretory bodies and mitochondria was lost. The T2 secretory bodies were also found altered in the tegumental cell bodies, suggesting that the alterations started at the production stage. Mitochondria were severely degenerated and located in the apical area of the tegument. The digestive system displayed a strong contraction, which included the disappearance of the intracecal lumen.

Praziquantel efficacy against Brachylaima sp. metacercariae (Trematoda: Brachylaimidae) parasitizing the edible landsnail Cornu aspersum and its HPLC-MS/MS residue determination.

Cornu aspersum is a terrestrial edible snail, often parasitized by Brachylaima (Trematoda) metacercariae. Ingestion of undercooked snails by humans allows metacercariae to develop to adult in the intestine causing brachylaimiasis (expected mortality rate 5-10%). The cosmopolitan character of the trematode and of its vector, enhanced in a future climate change scenario, and the absence of adequate sanitary controls of the snails in marketplaces clearly increase the risk of human brachylaimiasis. The treatment of farmed snails with praziquatel (PZQ) would be a tool to control this food-borne disease. The objectives of this study were: to report the prevalence of Brachylaima metacercariae parasitizing C. aspersum in marketplaces, to assess the efficacy and tolerance of PZQ in C. aspersum and to develop an HPLC-MS/MS analytical method to quantify PZQ residue in the edible snail (the acceptable daily intake of PZQ is 0.17 mg/kg bw in humans). Prevalence of parasitization of C. aspersum by Brachylaima metacercariae in public marketplaces reached the 80%. PZQ was orally administered to snails, mixed with the usual snail food. In dose determination assays three doses were individually tested (10 days): 1.2 mg PZQ/snail, 1.8 mg PZQ/snail (efficacy 97.3%, p < 0.001) and 2.4 mg PZQ/snail (efficacy 98.7%, p < 0.001). In dose confirmation tests (n = 200) the 2.4 mg PZQ/snail dose was group tested (10 snails/box, 7 days): 2 g of PZQ supplemented snail food were disposed daily in the treatment group boxes and 2 g of snail food (placebo) in the control boxes (efficacy 94.6%, p < 0.001; prevalence dropped from 68.7% in control group to 10.1% in treatment group, p < 0.001). Received dose was 220.2 mg PZQ/kg snail with shell. In the analytical method, linearity, lower limit of quantification (0.05 µg/ml), selectivity, carry over, accuracy, precision, dilution integrity, matrix effect and stability were tested. Sixty snails were treated (11 mg PZQ/g snail food) and analyzed. PZQ was detected and quantified (0.093 mg PZQ/g snail). PZQ treatment of C. aspersum is effective, well tolerated by the snail, affordable and easy to reproduce in snail farms.

Prevalence and climatic associated factors of Cryptosporidium sp. infections in savanna chimpanzees from Ugalla, Western Tanzania.

Studies about parasitization by Cryptosporidium in great apes have been scarce and mostly conducted in captivity. The present study reports the presence of Cryptosporidium sp. in wild chimpanzees (Pan troglodytes) from Ugalla, western Tanzania. Ugalla is one of the driest, most open, and seasonal habitats inhabited by chimpanzees. Cryptosporidium sp. was found in 8.9 % of the samples. The presence of the parasite was determined by preserving fecal samples in chemical conventional fixatives (MIF and alcohol absolute) staining them using a modified Zielh-Neelsen technique, and examining them with a light microscope. The number of fecal samples positive for Cryptosporidium was significantly higher during the rainy than during the dry season (p < 0.005). The results showed that feces collected in the rainy season were almost three times more likely to be positive for Cryptosporidium than those collected in the dry season (OR = 2.81). Cryptosporidium detection was significantly negatively affected by highest temperatures (>28.7 °C, p < 0.001). Cryptosporidiosis can cause serious health problems in humans and its potential effect on Ugalla chimpanzees is discussed.

Prevalence and associated factors of intestinal parasitisation: a cross-sectional study among outpatients with gastrointestinal symptoms in Catalonia, Spain.

The aim of this study was to report the prevalence of intestinal parasites in stool specimens from outpatients in Catalonia (Spain), and to evaluate the association of age, seasonality, and gender on general parasitisation and by the most frequent detected species. A total of 13,913 samples from 8,313 patients (1-3 specimens per patient) reporting digestive disorders were examined between 1999 and 2005 as a part of medical examinations. Samples were fixed with MIF solution and microscopically examined as wet mounts. Permanent stain was obtained by the modified Ziehl-Neelsen technique for intestinal coccidia. Nineteen species of intestinal parasites were identified. Blastocystis hominis (585 patients) was the predominant species, followed by Giardia duodenalis (321), Dientamoeba fragilis (131), Entamoeba coli (60) and Cryptosporidium sp. (59). Prevalence of helminths was low, being Enterobius vermicularis as the most frequently reported helminth (49 patients). The overall parasitisation was 1,136/8,313 (13.7%); prevalence in adults was 19.8% with a maximum in spring (14.8%). In the adjusted models, age was the main factor associated with infection: adults, with B. hominis and Entamoeba coli (odds ratio (OR) = 6.0 and OR = 8.5, respectively) and children, with Cryptosporidium and Giardia (OR = 2.0 and OR = 3.3, respectively). However, seasonality cannot be considered related with infection. The total prevalence was low, taking into account that all the subjects examined presented gastrointestinal symptoms and that species traditionally considered as non-pathogenic were included in the study.

Parásitos intestinales en inmigrantes atendidos durante el 2007 en una Unidad de Medicina Tropical de Barcelona / No disponible

Introducción: El aumento de inmigración procedente de países de baja renta puede contribuir al aumento de ciertas enfermedades infecciosas, especialmente aquellas de distribución cosmopolita. Actualmente, existen pocos datos referentes al estado de salud de la población recién llegada. El objetivo fue determinar la prevalencia de parásitos intestinales en inmigrantes y describir la influencia de variables sociodemográficas y clínicas en la parasitación. Material y métodos: Estudio descriptivo de 173 muestras fecales de pacientes con y sin sintomatología digestiva, sometidos a cribado parasitológico en una unidad de medicina tropical de Barcelona, entre el 1 de enero y el 31 de mayo del 2007.Resultados: La prevalencia de parasitación fue del 52,6%. Se detectaron 8 especies de protozoos, 7 de helmintos y Blastocysitis hominis, pero ningún caso de Cryptosporidium sp. Los inmigrantes recién llegados (p=0,026) y los procedentes de África y Asia resultaron los más parasitados (p=0,017). Los pacientes con helmintos, especialmente frecuentes entre los africanos (p=0,004), presentaron elevados valores de eosinófilos (p<0.001). El 45,0% de pacientes atendidos eran portadores asintomáticos. Conclusiones: A pesar de la dificultad de diseminación en nuestro medio, las elevadas tasas detectadas de parasitación intestinal justificarían el cribado parasitológico sistemático en pacientes asintomáticos procedentes de países de baja renta (AU)

Background: Increase of immigrant population proceeding from low-income countries could contribute to the emergence of some infectious diseases, especially those with cosmopolitan distribution. Nowadays, scarce data are known about health situation of newly arrived immigrants. The aim of the study was to determine the prevalence of intestinal parasites in immigrants and to describe the influence of demographic and clinical factors in parasitation. Material and methods: A descriptive study was carried out among 173 faecal samples from patients with and without digestive disorders, screened in a tropical diseases unit in Barcelona, from 1st January to 31st May 2007.Results: The prevalence of parasitation was 52,6%. 8 protozoa species, 7 helminth species and Blastocystis hominis, have been identified. No positive samples of Crypstosporidium sp. were found. Newly arrived patients (p=0,026) and those proceeding from Africa and Asia werethe most parasitized (p=0,017). Helminths parasitation was mainly found in African patients(p=0,004) and those with high levels of eosinophils (p<0,001). 45,0% of patients were asymptomaticcarriers. Conclusions: In spite of the difficulty of dissemination in our country, the high rates of intestinal parasitation detected would justify systematic screening in asymptomatic patients proceeding from low-income countries (AU)

Life cycle and description of a new species of brachylaimid (Trematoda: Digenea) in Spain.

The life cycle of Brachylaima llobregatensis n. sp. (Trematoda: Brachylaimidae) is elucidated. Embryonated, operculated, asymmetric eggs (30.9 x 18.2 microm) are eliminated with feces of natural hosts wood mouse Mus spretus; white-toothed shrew, Crocidura russula; and an experimental host, domestic mouse, Mus musculus var. domesticus. The eggs are ingested by the helicid gastropod Helix (Cornu) aspersa, the only natural and experimental first intermediate host. The miracidium hatches from the egg, infects the snails, and develops into a branched sporocyst in the digestive gland. Microcaudate cercariae emerge from this snail and develop into unencysted metacercariae in the kidney of second intermediate host snails H. (C.) aspersa and Otala punctata (natural hosts) and Theba pisana (experimental host). Ingestion of infected snails leads to the infection of definitive hosts, with the adults inhabiting the middle part of the small intestine. There is a chaetotaxic pattern specific on the acetabular (S(II) 5-6 papillae) and body (papillae absent on P(II)) levels. Three types of cercaria papillae were observed by scanning electron microscopy: argentophilic papillae with fingerlike processes (cephalic, body, and acetabular levels); argentophilic papillae with opening (2 papillae in the M body level); and nonargentophilic dome-shaped papillae (on the cephalic C(II) level, alternating with argentophilic S(II) papillae on the ventral sucker). Suckers are subequal, with the acetabulum located in the posterior part of the anterior third of body. Vitellaria extend from anterior margin of acetabulum to between middle level and anterior margin of anterior testis.

Life cycle of Brachylaima mascomai n. sp. (Trematoda: Brachylaimidae), a parasite of rats in the Llobregat delta (Spain).

The terrestrial triheteroxenous life cycle of Brachylaima mascomai n. sp. (Trematoda: Brachylaimidae) is elucidated. Operculated, assymetric, embryonated eggs (25.4 x 12.7 microm) are passed with feces of the natural (Rattus norvegicus and R. rattus) and experimental (albino and wild mice, albino rats, Apodemus sylvaticus, Mus spretus [Muridae] and the golden gerbil) definitive hosts and ingested by the helicid gastropod Pseudotachea splendida, the only natural and experimental first intermediate host. Microcaudate cercariae harbored in branched sporocysts in the digestive gland emerge from this snail and contact P. splendida, Otala punctata, Theba pisana, and Helix (C.) aspersa snails developing into unencysted infective metacercariae in the kidney. Definitive hosts are infected by ingestion of infected snails; the adult parasites inhabit the small intestine. Chaetotaxic cercarial pattern specific at acetabular (S(II) 8-10 papillae) and cephalic (C(III) 13-15 papillae, H 16 papillae) levels. Three types of cercarial papillae are observed by scanning electron microscopy (SEM) and their arrangement is correlated with chaetotaxy for the first time in trematodes: argentophilic papillae with fingerlike process (cephalic, body, and acetabular levels), argentophilic papillae with opening (2 papillae in the M body level), and nonargentophilic dome-shaped papillae (alternated with argentophilic S(II) papillae on the ventral sucker). SEM detected interlacing network of ridges covering the metacercarial body. Adults with multidigitate tegumentary spines were observed by SEM. Subequal suckers; the acetabulum located in the posterior part of anterior fifth of body. Vitellaria extend from between middle level and anterior margin of anterior testis to between middle level and posterior margin of acetabulum. Uterus almost reaches the intestinal bifurcation.