The association of urbanicity and travel time with lung cancer screening utilization.

BACKGROUND: To examine 1) the rate of lung cancer screening (LCS) utilization in a large healthcare system in South Carolina; 2) associations of urbanicity and travel time with LCS utilization. METHODS: LCS-eligible patients from 2019 were identified. The outcome was LCS utilization. The exposures were zip-code level urbanicity and travel time from the centroid of zip-code area to the nearest screening site (<10,10-<20, ≥20 min). Covariates included age, sex, race, marital status, insurance, body mass index, chronic obstructive pulmonary disease, Charlson Comorbidity Index (0, 1, 2, ≥3), and zip-code level median income. Chi-square tests and logistic regressions were employed. RESULTS: The analysis included 6930 patients, among whom 1432 (20.66%) received LCS. After adjusting for covariates, living in a non-metropolitan area (adjusted odds ratio: 0.32; 95% confidence interval: 0.26-0.40) and having longer travel time (0.80 [0.65-0.98] and 0.68 [0.54-0.86] for 10-<20 and ≥20 min travel time, respectively, compared to <10 min travel time) were significantly associated with lower odds of LCS utilization. CONCLUSIONS: The LCS utilization rate of a healthcare system was about 20% in 2019. Living in non-metropolitan areas or having longer travel time to LCS site were associated with lower LCS utilization.

Molecular characterization of waterborne protozoa in surface water and sediment in Brazil: a taxonomic survey of ciliated protozoa and their correlation with Giardia duodenalis and Cryptosporidium spp.

The detection of Giardia duodenalis and Cryptosporidium spp. was performed, along with the identification of the ciliated protozoa biodiversity, to evaluate the correlation between these protozoa in freshwater quality monitoring. Water and sediment samples from two sites in the Atibaia River (Campinas, São Paulo, Brazil) were collected monthly for 2 years (n = 96). Pathogenic protozoa in water and sediment were detected by using immunomagnetic separation, followed by visualization by immunofluorescence assay (IFA). All positive aliquots in IFA were subjected to DNA extraction and subsequently nested PCR. Qualitative (in vivo observation and silver impregnation) and quantitative (in vivo enumeration) analyses were performed for the ciliated protozoa. Giardia cysts were detected in 62.5% of the surface water samples and Cryptosporidium spp. in 25.0%. In the sediment, cysts were detected in 35.4% samples and oocysts in 16.6%. A total of 57 samples positive for Giardia cysts were subjected to sequencing, 40 of which were harboring G. duodenalis (24 were characterized as sub-assemblage AII). For ciliated protozoa, 73 taxa belonging to 53 genera were identified over the period of the study. These results revealed a high degree of contamination by waterborne protozoa in the main water source which supplies drinking water for more than one million people in Campinas (São Paulo), highlighting the need for continuous monitoring of this catchment site. In addition, the present study provides important data regarding the sources of the water body degradation, i.e., fecal contamination of human origin, in addition to the survey of the ciliated protozoa.

A 'novel' multi-component approach to promote physical activity among older cancer survivors: a pilot randomized controlled trial.

BACKGROUND: Physical activity (PA) provides many benefits for recovery from cancer treatments. Many older (65+ years) cancer survivors which comprise the majority of the cancer survivor population, do not meet recommended PA guidelines. This study explored the feasibility and acceptability of using audiobooks as part of a 12-week multi-component intervention to increase steps/day, light and moderate-to-vigorous intensity PA among older survivors. METHODS: Twenty older cancer survivors (95% female, mean age = 71.55 years, 90% White, 85% overweight/obese, 75% breast cancer survivors, mean 1.96 years since treatment completion) were randomized into one of the two study groups (Audiobook Group, n = 12, Comparison Group, n = 8). Both study groups were provided a tailored step goal program over the 12-week intervention; weekly step increases were based on a percent increase from baseline. Participant self-monitored their steps using a Fitbit Charge 2. In addition, the Audiobook group were encouraged to listen to audiobooks (downloaded onto a smartphone device via an app available at no cost from the local library) during PA to add enjoyment and increase PA. Regression analyses on steps/day, light and moderate-to-vigorous PA/week and sedentary time/week as assessed by the Actigraph were conducted, after adjusting for Actigraph wear time. Data from the post-intervention questionnaire were summarized. RESULTS: Overall, majority of participants (89%) stated they were very satisfied with their participation and 100% reported that they were able to maintain their activity upon study completion. Retention rates were high. At post-intervention, there were significant differences favoring the Audiobook group for steps/day and moderate-to-vigorous PA/week. No significant group differences were found for minutes of light intensity PA/week and sedentary time/week. CONCLUSION: Piloting the implementation of a sustainable, innovative intervention among older survivors to increase their PA has significance for this group of survivors.

Improving Adherence to Adjuvant Hormonal Therapy Among Disadvantaged Women Diagnosed with Breast Cancer in South Carolina: Proposal for a Multimethod Study.

BACKGROUND: Current clinical guidelines recommend that hormone receptor-positive breast cancer survivors take adjuvant hormonal therapy (AHT) for 5 to 10 years, following the end of definitive treatment. However, fewer than half of patients adhere to the guidelines, and suboptimal adherence to AHT is associated with an increased risk of breast cancer mortality. Research has extensively documented sociodemographic and disease-specific factors associated with adherence to AHT, but very little evidence exists on behavioral factors (eg, knowledge, patient-provider communication) that can be modified and targeted by interventions. OBJECTIVE: The goal of this study is to develop and test a theory-based, multilevel intervention to improve adherence to AHT among breast cancer survivors from racially and socioeconomically disadvantaged backgrounds (eg, Medicaid-insured). The specific aims are to (1) explore multilevel (eg, patient, health care system) factors that influence adherence to AHT; (2) develop a theory-based, multilevel intervention to improve adherence to AHT; and (3) pilot test and evaluate the intervention developed in Aim 2. METHODS: For Aim 1, we will recruit breast cancer survivors and health care professionals to participate in semistructured interviews to gain their perspectives about barriers and facilitators to AHT use. We will conduct a directed content analysis of the Aim 1 qualitative interview data. For Aim 2, we will integrate Aim 1 findings and current literature into the design of a multilevel intervention using an Intervention Mapping approach. For Aim 3, we will recruit Medicaid-insured breast cancer survivors to assess the feasibility of the pilot intervention. RESULTS: From May 2016 to July 2018, we completed interviews with 19 breast cancer survivors and 23 health care professionals in South Carolina. We will conduct a directed content analysis of the qualitative interview data. Results from this analysis will be used, in combination with current literature, to design (Aim 2) and pilot test a theory-based multilevel intervention (Aim 3) in Summer 2021. Results of the pilot are expected for Fall 2021. CONCLUSIONS: This study will provide a deeper understanding of how to improve adherence to AHT, using a novel and multilevel approach, among socioeconomically disadvantaged breast cancer survivors who often experience disproportionate breast cancer mortality. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/17742.

Toxoplasmagondii oocysts, Giardia cysts and Cryptosporidium oocysts in outdoor swimming pools in Brazil.

The accidental ingestion of treated recreational water is an important transmission route of waterborne protozoa worldwide. The present study aimed to provide the first evaluation of swimming pools in Brazil, analysing the presence of pathogenic protozoa (Toxoplasma gondii, Cryptosporidium spp. and Giardia spp.) by parasitological and molecular methods. A total of 57 samples were collected from 21 public swimming pools, either directly from the pool or filter backwash water and concentrated using the membrane filtration technique. Giardia cysts and Cryptosporidium oocysts were visualized by direct immunofluorescence assay after purification by immunomagnetic separation. Toxoplasma gondii oocysts were detected by autofluorescence visualization using the supernatant discarded during the purification step as a sample. Positive samples were submitted to molecular analysis. The molecular markers were used: SSU-rRNA, tpi, gdh and bg, for Giardia DNA amplification, and 18S rRNA gene fragment amplification was used for the Cryptosporidium oocysts. The 529-bp repeat element (REP529) fragment and the 35-fold repetitive B1 gene were employed as a target for T. gondii. Amplified products were submitted to sequencing and phylogenetic analysis. Giardia cysts were detected in 19.0% and Cryptosporidium oocysts in 9.5% of swimming pools. In one swimming pool (4.7%), both protozoa were detected on at least one occasion. Structures similar to T. gondii oocysts were detected in 33.3% of the samples, ranging from one to 23 per slide. Giardia was confirmed by DNA amplification in three swimming pools; Giardia duodenalis Assemblage A was identified by the phylogenetic positioning of the ß-giardin gene. Toxoplasma gondii DNA was detected in 14.2% of swimming pools. The present study represents the first report of the occurrence of T. gondii oocysts in swimming pools. Recreational activity in swimming pools contaminated by chlorine-resistant protozoa can represent a high risk of infection for bathers and swimmers.

Symptom Management Among Cancer Survivors: Randomized Pilot Intervention Trial of Heart Rate Variability Biofeedback.

Chronic cancer-related symptoms (stress, fatigue, pain, depression, insomnia) may be linked with sympathetic nervous system over-activation and autonomic imbalance. Decreased heart rate variability (HRV) is an indicator of autonomic dysregulation that is commonly observed among cancer survivors. HRV biofeedback (HRVB) training induces HRV coherence, which maximizes HRV and facilitates autonomic and cardiorespiratory homeostasis. This randomized, wait-list-controlled, pilot intervention trial tested the hypothesis that HRVB can improve HRV coherence and alleviate cancer-related symptoms. The intervention group (n = 17) received 4-6 weekly HRVB training sessions until participants demonstrated skill acquisition. Controls (n = 17) received usual care. Outcomes assessed at baseline and follow-up included 15-min HRV recordings (HRV Coherence Ratio), and symptoms of: stress, distress, post-traumatic stress disorder (PTSD), pain, depression, fatigue, and sleep disturbance. Linear mixed models for repeated measures were used to assess Group-by-Time interactions, pre- versus post-treatment differences in mean symptom scores, and group differences at follow-up. Mean HRV Coherence Ratios (± standard error) improved in the HRVB group at follow-up (baseline: 0.37 ± 0.05, post-intervention: 0.84 ± 0.18, p = 0.01), indicating intervention validity. Statistically significant Group-by-Time interactions indicated treatment-related improvements in HRV Coherence Ratios (p = 0.03, Pre-vs. post-treatment effect size [Cohen's d]: 0.98), sleep symptoms (p = 0.001, d = 1.19), and sleep-related daytime impairment (p = 0.005, d = 0.86). Relative to controls, the intervention group experienced trends toward improvements in stress, distress, fatigue, PTSD, and depression, although no other statistically significant Group-by-Time interactions were observed. This pilot intervention found that HRVB training reduced symptoms of sleep disturbance among cancer survivors. Larger-scale interventions are warranted to further evaluate the role of HRVB for managing symptoms in this population. Registration: NCT03692624 www.clinicaltrials.gov.

Occurrence and diversity of ciliated protozoa in the most commonly consumed leafy greens in Brazil: a qualitative study

Ciliated protozoa are an ubiquitous group of eukaryotes that have been poorly reported on leafy greens. The present study aimed to verify the occurrence and diversity of ciliated protozoa in the leaves and roots of three of the most commonly consumed leafy greens in Brazil ­ lettuce, rocket and coriander. The vegetable samples were washed by manual agitation (3 minutes) in two different media (mineral water and Page ́s Amoeba Saline solution). After washing, the contents were incubated in Petri dishes and aliquots were removed for microscopic identification and in vivo observation. A total of 21 ciliated protozoa species were found, most of which were bacterivorous. Leafy greens have commonly been associated with foodborne outbreaks and ciliated protozoa, which although they are not a Public Health concern, can act as "Trojan Horses" harboring bacteria, viruses and other protozoa cysts and oocysts and can suggest a new route towards microbiological quality related to the food chain. This is the first report of ciliated protozoa on leafy greens consumed in Brazil. (AU)

Os protozoários ciliados constituem um grupo onipresente de eucariotos pouco relatados em vegetais folhosos. O presente estudo teve como objetivo verificar a ocorrência e a diversidade de protozoários ciliados nas folhas e raízes de três dos vegetais folhosos mais consumidos no Brasil - alface, rúcula e coentro. As amostras de vegetais foram lavadas por agitação manual (3 minutos) em dois meios diferentes (água mineral e Solução Salina para Ameba). Após a lavagem, o conteúdo foi incubado em placas de Petri e alíquotas foram retiradas para identificação microscópica e observação in vivo dos organismos. Um total de 21 espécies de protozoários ciliados foi encontrado, a maioria das quais era bacterívora. Tais hortaliças têm sido comumente associadas a surtos transmitidos por alimentos e os protozoários ciliados que, embora não sejam um problema de saúde pública, podem atuar como "Cavalos de Tróia" ao abrigar bactérias, vírus além de cistos e oocistos de protozoários o que pode sugerir uma nova rota para a avaliação da qualidade microbiológica relacionada à cadeia alimentar. Este é o primeiro relato de protozoários ciliados em folhas verdes consumidas no Brasil. (AU)

Occurrence and diversity of ciliated protozoa in the most commonly consumed leafy greens in Brazil: a qualitative study / Ocorrência e diversidade de protozoários ciliados nos vegetais folhosos mais consumidos no Brasil: um estudo qualitativo

Ciliated protozoa are an ubiquitous group of eukaryotes that have been poorly reported on leafy greens. The present study aimed to verify the occurrence and diversity of ciliated protozoa in the leaves and roots of three of the most commonly consumed leafy greens in Brazil – lettuce, rocket and coriander. The vegetable samples were washed by manual agitation (3 minutes) in two different media (mineral water and Page ́s Amoeba Saline solution). After washing, the contents were incubated in Petri dishes and aliquots were removed for microscopic identification and in vivo observation. A total of 21 ciliated protozoa species were found, most of which were bacterivorous. Leafy greens have commonly been associated with foodborne outbreaks and ciliated protozoa, which although they are not a Public Health concern, can act as “Trojan Horses” harboring bacteria, viruses and other protozoa cysts and oocysts and can suggest a new route towards microbiological quality related to the food chain. This is the first report of ciliated protozoa on leafy greens consumed in Brazil.

Os protozoários ciliados constituem um grupo onipresente de eucariotos pouco relatados em vegetais folhosos. O presente estudo teve como objetivo verificar a ocorrência e a diversidade de protozoários ciliados nas folhas e raízes de três dos vegetais folhosos mais consumidos no Brasil - alface, rúcula e coentro. As amostras de vegetais foram lavadas por agitação manual (3 minutos) em dois meios diferentes (água mineral e Solução Salina para Ameba). Após a lavagem, o conteúdo foi incubado em placas de Petri e alíquotas foram retiradas para identificação microscópica e observação in vivo dos organismos. Um total de 21 espécies de protozoários ciliados foi encontrado, a maioria das quais era bacterívora. Tais hortaliças têm sido comumente associadas a surtos transmitidos por alimentos e os protozoários ciliados que, embora não sejam um problema de saúde pública, podem atuar como “Cavalos de Tróia” ao abrigar bactérias, vírus além de cistos e oocistos de protozoários o que pode sugerir uma nova rota para a avaliação da qualidade microbiológica relacionada à cadeia alimentar. Este é o primeiro relato de protozoários ciliados em folhas verdes consumidas no Brasil.

Giardia spp. and Cryptosporidium spp. removal efficiency of a combined fixed-film system treating domestic wastewater receiving hospital effluent.

Giardia and Cryptosporidium have caused numerous outbreaks of diarrhea as a result of the ingestion of water contaminated with sewage. In Brazil, the efficiency of Giardia and Cryptosporidium removal by combined fixed-film systems has rarely been studied. The aims of the present study were therefore to verify the removal efficiency of Giardia and Cryptosporidium by a combined system (anaerobic/anoxic filter and aerated submerged biofilter) and to perform the genetic characterization of these parasites. The (oo)cysts were detected by centrifuge concentration and membrane filtration from raw sewage, effluents, adhered biomass, and sludge samples. Immunofluorescence assay and differential interference contrast microscopy were used for the visualization of the (oo)cysts. Nested PCR was applied to confirm Giardia and Cryptosporidium. Giardia and Cryptosporidium were detected in 27% and 5.5% of the 144 analyzed samples of raw sewage and effluents, respectively. A total of 33,000 cysts/L were recovered in the adhered biomass samples (n = 25) from different points of the aerated submerged biofilter, while 6000 oocysts/L were registered in a single point. An average of 11,800 cysts/L were found in the sludge samples (n = 5). The combined system exhibited a removal efficiency of Giardia cysts of 1.8 ± 1.0 log removal. The C and BIV assemblages of Giardia were identified in the raw sewage while AII was found in the treated effluent sample. It was not possible to calculate the removal efficiency of Cryptosporidium oocysts by the combined system. The combined system exhibited some potential as a suitable treatment for the removal of parasites from sewage.

Randomized Controlled Trial of a 4-Week Mindfulness Intervention among Cancer Survivors Compared to a Breathing Control.

Purpose: This randomized controlled trial (RCT: NCT02633748) examined effectiveness of an abbreviated mindfulness program on psychosocial symptoms, behaviors, and inflammation. Methods: Cancer survivors (n = 36) underwent a 4-week mindfulness intervention compared to a breathing control. Data included psychosocial questionnaires, anthropometrics, actigraphy, and blood draws. Clinic visits occurred at baseline, post-intervention, and a 3-month post-follow-up. Results: Compared to baseline, the intervention arm displayed reductions in sedentary time and perceived stress and improvements in subjective sleep quality, and daily steps at post-follow-up. Conclusions: An abbreviated mindfulness intervention in feasible in cancer survivors and signs of efficacy warrant further mindfulness studies among this population.

Pre-ozonation of source water: Assessment of efficacy against Giardia duodenalis cysts and effects on natural organic matter.

The protozoan Giardia duodenalis is one of the parasites of primary concern in drinking water treatment plants, due to its resistance to chlorination. Another matter of concern regarding chlorination of drinking water is the formation of disinfection by-products in the presence of precursors such as natural organic matter (NOM). In this study, the effects of ozonation (5 mg L-1) on G. duodenalis cysts in raw surface water from a drinking water treatment plant were evaluated, and the presence and alteration of NOM were assessed, as an indicative of the potential to prevent total organic halogen (TOX) formation during post-chlorination. Following ozone treatment, the presence of damaged cysts was observed by direct immunofluorescence assay (IFA) and confirmed by the molecular approach propidium monoazide-polymerase chain reaction (PMA-PCR). Using an animal model, analysis of the intestinal tissues revealed that 80% of the animals inoculated with ozonated water were positive for trophozoites. This study shows that analysis of intestinal fragments is imperative to accurately assess animal infection following inoculation of treated cysts. More importantly, considering the low infective dose of Giardia cysts in susceptible hosts, an ozone dosage usually applied in drinking water treatment plants did not completely inactivate G. duodenalis cysts in surface water. Nonetheless, the results suggest that competitive reactions with NOM have occurred, and the applied ozone dosage has proven useful to remove NOM reactivity, and thus prevent halogenated DBP formation during post-chlorination.

Molecular genotyping, diversity studies and high-resolution molecular markers unveiled by microsatellites in Giardia duodenalis.

BACKGROUND: Giardia duodenalis (synonyms G. lamblia and G. intestinalis) is an enteric protozoan parasite of a wide range of mammalian hosts, including humans and various domestic and wild animals. There is considerable genetic variability in G. duodenalis and isolates of this parasite have been divided into eight genetic assemblages. Microsatellites markers can be used to discriminate isolates with a high level of sensitivity. This study was conducted to identify and characterize genomic microsatellites (simple sequence repeats-SSRs), sequences of one- to six-nucleotide motifs repeated in tandem, present in the available genomes of G. duodenalis and to develop new markers that can serve as a tool for detection and for characterizing the genetic diversity of this parasite. METHODOLOGY/ PRINCIPAL FINDINGS: For each genetic assemblage, polymorphism levels for the microsatellite markers were evaluated. After performing the analysis using the MISA and SciRoKo software, 1,853 simple sequence repeats (SSRs) were identified. In all the genomes, trinucleotide repeats were the most common class followed by tetranucleotide. Many of the SSR loci are assemblage-specific, and 36 SSR loci shared among all the genomes were identified. Together with hypothetical proteins, variant-specific surface proteins represented nearly half of the annotated SSR loci. The results regarding the most common repeat among the SSRs led us to infer that positive selection occurred to avoid frameshift mutations. Additionally, based on inter- and intra-genetic assemblages polymorphism analyses, we unveiled previously undetected genetic variation, indicating that the microsatellite markers we developed are useful molecular tools for epidemiological inferences based on population genetics patterns and processes. CONCLUSIONS: There is increasing demand for the development of new molecular markers and for the characterization of pathogens at a higher resolution level. In this study, we present 60 G. duodenalis microsatellites markers that exhibited high polymerase chain reaction (PCR) amplification efficiency among the different genetic assemblages. Twenty of these markers presented nucleotide sequence polymorphisms and may be used as a genotyping tool. The monomorphic markers can be used for the detection of the parasite at the species and genetic assemblage level. These polymorphic markers revealed a genetic diversity that was previously undetectable, thus they can be considered valuable molecular tools for high resolution markers in future studies investigating Giardia and may also be used for epidemiological inferences based on populations genetics patterns and processes.

Genotypic characterization and assessment of infectivity of human waterborne pathogens recovered from oysters and estuarine waters in Brazil.

Waterborne, food-borne and sewage-borne pathogens are a major global concern, with the annual recurrence, most notably during the summer, of outbreaks of gastroenteritis of unconfirmed etiology associated with recreational activities in marine environments. The consumption of contaminated water-based foodstuffs is also related to outbreaks of human illness. The main goals of the present study were: i) to identify the genetic assemblages of Giardia duodenalis cysts in growing and depurated oysters destined for human consumption on the southern coast of São Paulo, Brazil; ii) to verify the main circulating G. duodenalis assemblages and their subtypes in different brackish waters used for the production of mollusks and for recreational purposes; iii) to track the contamination of growing and depurated oysters by the human adenovirus and identify the infectivity of adenoviral particles recovered from oysters before and after depuration; iv) to evaluate the occurrence and genotype of the free-living amoebae of the genus Acanthamoeba in brackish water and oysters from all the sites described above. Four sampling sites in the Cananeia estuary were selected to search for pathogenic and amphizoic protozoa (Giardia and Acanthamoeba respectively): site 1: oyster growth, site 2: catchment water (before UV depuration procedure), site 3: filter backwash (filtration stage of water treatment) and site 4: oyster depuration tank. Oysters at sites 1 and 4 were evaluated for the presence of adenovirus (HAdV). Analysis consisted of conventional microbiological as well as molecular methods. Giardia duodenalis were detected in all the water sites analyzed and the molecular analysis revealed that sub-assemblage AII was the most frequently distributed throughout the estuarine environment, although one sample was identified as belonging to the assemblage C. Acanthamoeba were also isolated from different locations of the estuarine area, and were detected at all the analyzed sites. The majority of isolates belonged to the T3 genotype, while the T4 genotype was identified once. The sequencing reaction of Giardia duodenalis revealed the contamination of three batches of depurated oysters by the sub-assemblage AII. With respect to viruses, seven batches of oysters (four growing and three depurated) were found to be harboring infectious HAdV particles when submitted to plaque assay. Overall, the results of the sequencing reactions combined with the plaque assay revealed that the isolates of Giardia duodenalis and the infectious HAdV particles identified in oyster tissues have the potential to infect humans and pose a threat if consumed raw or lightly cooked. This is the first report on the sub-assemblage AII identified in oysters which are submitted to a cleaning and disinfection procedure prior to human consumption in Brazil. Acanthamoeba specific genotypes were also identified for the first time in a recreational estuarine area in Brazil, contributing to knowledge of their molecular and environmental epidemiology, which is considered scarce even in marine and estuarine areas of the world.

Occurrence and molecular characterization of Giardia duodenalis cysts and Cryptosporidium oocysts in raw water samples from the Rímac River, Peru.

Giardia and Cryptosporidium are potentially pathogenic protozoa which are ubiquitous in ambient surface water. The present study included 60 samples of surface water from three sampling sites from the Rímac River, Lima and Callao, Peru, to detect the occurrence of Giardia spp. and Cryptosporidium spp. and to perform molecular characterization of specimens found. Water samples were concentrated using the membrane filtration technique, and following elution, cysts and oocysts were visualized by direct immunofluorescence assay (IFA). For molecular characterization, tpi and bg gene fragments and 18S rRNA were amplified by nested PCR for Giardia and Cryptosporidium, respectively, followed by sequencing and phylogenetic analysis. Giardia cysts were found in 93.3% of the analyzed samples, whereas Cryptosporidium oocysts were detected in 15%. The positivity of the Giardia cysts was 86.6% (n = 26) in 2014, while Cryptosporidium oocysts were not detected. In 2015, both protozoa were found in raw water samples, with all 30 samples collected positive for Giardia cysts (100.0%) and 9 positive for Cryptosporidium oocysts (30.0%). Oocysts were detected in 20.0% of water samples from sites 1 (mean 5.25 oocysts/L) and 2 (mean 52.3 oocysts/L), while at site 3, oocysts were detected in 50.0% of raw water samples (mean 193.6 oocysts/L). The presence of Giardia duodenalis assemblage A was confirmed in several samples by the phylogenetic positioning of the bg and tpi genes, and the sub-assemblage AII was predominant (8/9). Sequencing for Cryptosporidium resulted in profiles compatible with Cryptosporidium hominis, Cryptosporidium meleagridis, and Cryptosporidium baileyi. This is the first time that the presence of G. duodenalis assemblage A/sub-assemblage AII and Cryptosporidium species has been reported in surface water samples in Peru. These Cryptosporidium species and the Giardia duodenalis assemblage are associated with human disease which highlights the potential risk to public health and the need to increase environmental monitoring measures to protect this water body.

Giardiasis as a neglected disease in Brazil: Systematic review of 20 years of publications.

INTRODUCTION: Giardiasis is an intestinal infection that affects more than two hundred million people annually worldwide; it is caused by the flagellated protozoan Giardia duodenalis. In tropical countries and in low or middle-income settings, like Brazil, its prevalence can be high. There is currently no systematic review on the presence of G. duodenalis in patients, animals or water sources in Brazil. METHODS: This systematic review was performed according to recommendations established by Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA). As databases for our searches, we have used PubMed, Embase, Scopus and the Brazilian database SciELO using the keywords «Giardia*¼ and «Brazil¼. RESULTS: This systematic review identified research studies related to G. duodenalis in water, giardiasis in animals, prevalence of giardiasis across Brazilian regions, genotyping of strains isolated in humans, and giardiasis in indigenous populations. We also propose a network of G. duodenalis transmission in Brazil based on genotypes analyses. CONCLUSION: This is the first time within the last twenty years that a review is being published on the occurrence of G. duodenalis in Brazil, addressing relevant issues such as prevalence, molecular epidemiology and analytical methods for parasite detection.

Enterocytozoon bieneusi detected by molecular methods in raw sewage and treated effluent from a combined system in Brazil.

BACKGROUND: Enterocytozoon bieneusi are the most common microsporidia associated with different clinical manifestations such as diarrhoea, respiratory tract inflammation and acalculous cholecystitis, especially in immunocompromised patients. Infection usually occurs by ingestion of food and water contaminated with spores, but can also result from direct contact with spores through broken skin, eye lesions, and sexual transmission, depending on the microsporidian species. Although there are reports of E. bieneusi found in humans and animals in Brazil, there are no published studies of environmental samples examined by molecular methods. OBJECTIVES: The purpose of this study was to verify the presence of E. bieneusi in raw sewage and treated effluent from a combined system by molecular methods. METHODS: Raw sewage and treated effluent samples collected from a combined system were analysed for the presence of E. bieneusi using the internal transcriber spacer (ITS) region of E. bieneusi by nested polymerase chain reaction. FINDINGS: The analysis revealed E. bieneusi presence and a novel genotype (EbRB) in one raw sewage sample and one treated effluent. MAIN CONCLUSIONS: The presence of E. bieneusi in final effluent indicates that the combined system may not remove microsporidian spores. This study is the first report of E. bieneusi in environmental samples in Brazil.

Enterocytozoon bieneusi detected by molecular methods in raw sewage and treated effluent from a combined system in Brazil

BACKGROUND Enterocytozoon bieneusi are the most common microsporidia associated with different clinical manifestations such as diarrhoea, respiratory tract inflammation and acalculous cholecystitis, especially in immunocompromised patients. Infection usually occurs by ingestion of food and water contaminated with spores, but can also result from direct contact with spores through broken skin, eye lesions, and sexual transmission, depending on the microsporidian species. Although there are reports of E. bieneusi found in humans and animals in Brazil, there are no published studies of environmental samples examined by molecular methods. OBJECTIVES The purpose of this study was to verify the presence of E. bieneusi in raw sewage and treated effluent from a combined system by molecular methods. METHODS Raw sewage and treated effluent samples collected from a combined system were analysed for the presence of E. bieneusi using the internal transcriber spacer (ITS) region of E. bieneusi by nested polymerase chain reaction. FINDINGS The analysis revealed E. bieneusi presence and a novel genotype (EbRB) in one raw sewage sample and one treated effluent. MAIN CONCLUSIONS The presence of E. bieneusi in final effluent indicates that the combined system may not remove microsporidian spores. This study is the first report of E. bieneusi in environmental samples in Brazil.

Population genetic analysis of Giardia duodenalis: genetic diversity and haplotype sharing between clinical and environmental sources.

Giardia duodenalis is a flagellated intestinal protozoan responsible for infections in various hosts including humans and several wild and domestic animals. Few studies have correlated environmental contamination and clinical infections in the same region. The aim of this study was to compare groups of Giardia duodenalis from clinical and environmental sources through population genetic analyses to verify haplotype sharing and the degree of genetic similarity among populations from clinical and environmental sources in the metropolitan region of Campinas. The results showed high diversity of haplotypes and substantial genetic similarity between clinical and environmental groups of G. duodenalis. We demonstrated sharing of Giardia genotypes among the different populations studied. The comparison between veterinary and human sequences led us to identify new zoonotic genotypes, including human isolates from genetic assemblage C. The application of a population genetic analysis in epidemiological studies allows quantification of the degree of genetic similarity among populations of Giardia duodenalis from different sources of contamination. The genetic similarity of Giardia isolates among human, veterinary, and environmental groups reinforced the correlation between clinical and environmental isolates in this region, which is of great importance for public health.

Ciliated protozoa community of a combined UASB-activated sludge system in southeastern Brazil.

The aims of the present study were (1) to evaluate the abundance and taxonomic composition of ciliated protozoa in the activated sludge of a full-scale combined anaerobic-aerobic system operating in a tropical country and (2) to study the relationship between the effluent quality, the physicochemical variables, and the ciliates present in the operating system. The total ciliate fauna of the activated sludge of the Piçarrão Wastewater Treatment Plant (Piçarrão WWTP) was composed of 36 morphospecies belonging to 33 genera. These included 21 species observed in the activated sludge samples on the day of collection and 15 species found in cultures. The activated sludge of the Piçarrão WWTP contained a diversified ciliate community composed mainly of indicator organisms. The most frequently occurring morphospecies were Aspidisca cicada, Vorticella spp., Gastronauta aloisi, Acineria uncinata, and Epistylis plicatilis complex. These results showed that satisfactory operating conditions prevailed at the Piçarrão WWTP. In the combined UASB-activated sludge system, the presence of Aspidisca cicada suggests the occurrence of denitrification in the process while the presence of Acineria uncinata and G. alosi indicates the removal of carbonaceous organic matter.

Real scale environmental monitoring of zoonotic protozoa and helminth eggs in biosolid samples in Brazil.

Biosolid is the product of the activated sludge treatment system and its final disposition is subject of ongoing discussion as this residue can therefore harbor a great number and variety of pathogens. This study was aimed to (1) monitor the presence of Giardia and Cryptosporidium in biosolid samples from a treatment plant in Campinas, SP, Brazil, (2) observe Giardia cyst wall morphological integrity in treated samples using scanning electron microscopy (SEM) and (3) verify the presence and viability of helminth eggs. Cysts were present in 33.3 % of the samples, whereas oocysts were detected in 8.3 %. Viable Ascaris sp. Toxocara sp. and similar to Trichuris sp. eggs were found through the use of Mexican Official Norm. Results demonstrate the difficulties inherent in working with biosolid as factors such as temperature, ionic strength and pH influenced the recovery of cysts and oocysts. Pores and ruptures were not observed in cyst wall visualized by SEM following 45 days of exposure to sunlight, only minimal morphological changes. These observations emphasize both the importance of adequate treatment of sewage sludge and the need to develop appropriate techniques for the detection of Giardia and Cryptosporidium in this type of sample. This is the first time that a study was done in a real scale for biosolid samples in determining the presence of pathogenic protozoa as Giardia and Cryptosporidium in Brazil, and also observed minimal cyst wall damage after sunlight treatment.