Surveillance of Giardia and Cryptosporidium in sewage from an urban area in Brazil / Monitoramento de Giardia e Cryptosporidium em esgoto de uma área urbana no Brasil

Abstract Cryptosporidium and Giardia are protozoan parasites that cause diarrhea in humans and animals. Molecular characterization of these pathogens in sewage may provide insight on their occurrence and prevalence in Brazil. This study aimed to investigate the presence of Giardia and Cryptosporidium in raw and treated sewage from Londrina, Paraná, Brazil. Samples were collected every two weeks during a year. Samples were concentrated, then DNA was extracted and subjected to a nested PCR targeting the Giardia 18S rRNA gene and the Cryptosporidium 18S rRNA gene. Species of Cryptosporidium were characterized by restriction fragment length polymorphism (RFLP). All raw sewage and 76% of the treated sewage were positive for Giardia; 84% of raw sewage samples and 8% of treated sewage were positive for Cryptosporidium. C. muris, C. hominis, C. baileyi, C. parvum and C. suis were detected in 100%, 19%, 9%, 9% and 4% of raw sewage, respectively. C. muris was the only species found in treated sewage. Multiple species of Cryptosporidium were present in 19.04% of the raw sewage. Treated sewage water can pose a threat to human health. The speciation of Cryptosporidium revealed the presence of non-common zoonotic species as C. suis and C. muris.

Resumo Cryptosporidium e Giardia são protozoários causadores de diarreia em animais e humanos. A caracterização molecular destes protozoários em esgoto pode prover dados ainda desconhecidos da ocorrência de espécies. O objetivo do presente estudo foi monitorar a ocorrência de Giardia e espécies de Cryptosporidium em esgoto bruto e tratado em uma estação de tratamento de esgoto (ETE) de Londrina, Paraná. Amostras de esgoto bruto e tratado foram coletadas no período de um ano, com periodicidade quinzenal. A ocorrência destes protozoários foi caracterizada por meio de concentração das amostras e posterior extração de DNA seguida de nested-PCR para amplificação de fragmentos dos genes 18S rRNA de Giardia e 18S rRNA de Cryptosporidium. A caracterização das espécies de Cryptosporidium foi realizada por meio de análise por polimorfismo de comprimento do fragmento de restrição (RFLP) dos produtos obtidos. Foram coletadas no total 25 amostras de cada, esgoto bruto e esgoto tratado. Para Giardia, todas as amostras de esgoto bruto e 76% das de esgoto tratado foram positivas. Cryptosporidium esteve presente em 84% das amostras de esgoto bruto e em 8% do tratado. No esgoto tratado foi encontrado apenas C. muris, já nas amostras de esgoto bruto foram encontradas cinco espécies: C. muris, C. hominis, C. baileyi, C. suis e C. parvum em 100%, 19%, 9%, 9% e 4%, respectivamente. A presença de espécies mistas foi observada em 19,04% das amostras. A presença de Giardia e Cryptosporidium em esgoto tratado pode pôr em risco a saúde humana. A discriminação de espécies de Cryptosporidium revelou a presença de espécies zoonóticas incomuns como C. suis e C. muris.

Identificação de helmintos encontrados em uma estação de tratamento de esgoto de Feira De Santana, Bahia, Brasil / Identification of helminths found in a sewage treatment plant in Feira De Santana, Bahia, Brazil / Identificación de helmintos encontrados en una estación de tratamiento de alcantarilla De Feira De Santana, Bahía, Brasil

O objetivo desse trabalho foi fazer a descrição morfológica e a identificação de Nematoda encontrados em uma estação de tratamento de esgoto de Feira de Santana, BA, Brasil. Foram coletadas 56 amostras de 500 mL em todos os pontos das etapas aeróbias do tratamento. As amostras foram processadas para estudo em microscopia óptica de campo claro (MO) e microscopia eletrônica de varredura (MEV). Foram encontradas larvas L1e L4, classificadas de acordo com o tamanho. As larvas L1 apresentaram as seguintes medidas de comprimento Md = 255,0?m; diâmetro Md = 15,0?m; vestíbulo bucal Md = 10,0?m; esôfago Md = 82,5?m; e cauda Md = 26,2?m. As larvas L4 apresentaram as seguintes medidas de comprimento Md = 740,0?m; diâmetro Md = 42,0?m; vestíbulo bucal Md = 22,5?m; esôfago Md = 200,0?m; e cauda Md = 47,5?m. Fêmeas adultas também foram encontradas, caracterizando-se por apresentam esôfago rabditoide, maturação sexual com presença de ovos na tuba uterina em diferentes fases de formação embrionária. Alguns ovos foram vistos larvados, sugerindo um padrão de larviparidade. Os vermes foram considerados de vida livre e classificadas como da Ordem Rhabditida, Família Strongyloidae e Gênero Strongyloides.

The objective of this work was to morphologically describe and identify Nematoda found in a sewage treatment plant in Feira de Santana, BA, Brazil. A total of 56 samples with 500-mL each were collected at all points in the aerobic treatment stages. The samples were processed for bright light microscopy (LM) and scanning electron microscopy (SEM). L1 and L4 larvae were found, classified according to size. L1 larvae presented the following measurements: Md=255.0-?m length; Md=15.0-?m diameter; Md=10.0-?m oral vestibule; Md=82.5-?m esophagus; and Md=26.2 ?m tail. L4 larvae presented the following measurements: Md=740.0-?m length; Md=42.0-?m diameter; Md=22.5-?m oral vestibule; Md=200.0-?m esophagus; and Md=47.5-?m tail. Adult females have also been found, characterized by rhabditoid esophagus, sexual maturation with presence of eggs in the uterine tube at different stages of embryo formation. Some eggs were seen at larvae stage, suggesting a pattern of larvae parity. The worms were considered as free-living and classified in the Rhabditida order, Strongyloidae Family and Strongyloides genus.

El objetivo de ese trabajo fue realizar la descripción morfológica y la identificación de Nematodo encontrados en una estación de tratamiento de alcantarilla de Feira de Santana, BAHIA, Brasil. Fueron recogidas 56 muestras de 500 mL en todos los puntos de las etapas aerobias del tratamiento. Las muestras fueron procesadas para estudio en microscopía óptica de campo claro (MO) y microscopia electrónica de barrido (MEB). Se encontraron larvas L1 y L4, clasificados según el tamaño. Las larvas L1 presentaron las siguientes medidas de longitud Md=255.0?m; diámetro Md=15.0?m; vestíbulo oral Md=10.0 µm; esófago Md=82,5?m; y cola Md=26.2?m. Las larvas L4 presentaron las siguientes medidas de longitud Md=740.0?m; diámetro Md=42.0?m; vestíbulo oral Md=22,5?m; esófago Md=200.0?m; y cola Md=47.5?m. También se han encontrado hembras adultas, caracterizándose por presentar esófago rabditoide, maduración sexual con presencia de huevos en las trompas uterina en diferentes etapas de la formación del embrión. Algunos huevos fueron vistos larvados, sugiriendo un estándar de larviparidad. Los gusanos fueron considerados de vida libre y clasificados como de Orden Rhabditida, Familia Strongyloidae y género Strongyloides.

Polymerase chain reaction and nested-PCR approaches for detecting Cryptosporidium in water catchments of water treatment plants in Curitiba, State of Paraná, Brazil

Introduction Cryptosporidium is an important protozoan cause of waterborne disease worldwide of concern to public health authorities. To prevent outbreaks of cryptosporidiosis, the monitoring of this parasite in drinking water is necessary. In the present work, the polymerase chain reaction (PCR) and nested-PCR techniques were used to detect Cryptosporidium in raw water from catchment points of four water treatment plants (WTP) in Curitiba, Paraná, Brazil. Methods First, DNA extraction techniques were tested in samples containing decreasing amount of oocysts in reagent water, and PCR and nested-PCR with specific primers for 18SSU rDNA of Cryptosporidium were conducted to determine their sensitivity. In reagent water, a commercial extraction kit provided the best analytical sensitivity, and PCR and nested-PCR allowed the detection of five and two oocysts, respectively, with the primers XIAOR/XIAOF and XIAO1F/XIAO2R. Results In the spiking experiments, only the PCR with the primers AWA995F/AWA1206R was successful at detecting concentrations of 0.1 oocysts/mL. Two catchments samples of raw water and/or water sludge from four WTPs were contaminated with Cryptosporidium. Conclusions The application of the techniques to monitor Cryptosporidium in water and detect contamination in water catchments of WTPs in Curitiba are discussed in the present work. .

Epidemiological Characteristics of the First Water-Borne Outbreak of Cryptosporidiosis in Seoul, Korea

The first case of human cryptosporidiosis was reported in Korea in 1995; however, an outbreak of Cryptosporidium has not been reported in Korea until now. This paper describes the first outbreak of cryptosporidiosis in Korea. On May 24, 2012, a local public health center filed a report on 126 residents with gastrointestinal symptoms in an old apartment complex in Seoul. Epidemiological investigations were implemented on 125 of the 126 patients. The patients were reported continuously over a period of 22 days. Diarrhea was the most common clinical symptom, and lasted for 5 days on average. The tap water was the only common exposure of the patients. During the environmental investigation it was discovered that the water and septic tanks were situated closely and that the waste water pipes were corroded where they passed over the water pipes. Cryptosporidium parvum was detected in 3 of the 7 stool specimens by PCR-RFLP. A number of Cryptosporidium oocysts were also detected in the water specimens from the water tank. In conclusion, Cryptosporidium parvum was the key causal pathogen of this outbreak. It is presumed that the tap water was contaminated by a sewage leak from the aged pipelines.

Recovery of gastrointestinal swine parasites in anaerobic biodigester systems / Recuperação de parasitas gastrintestinais de suínos em biodigestores anaeróbicos

Solid and liquid wastes from livestock operations represent important challenges for animal production regarding their impact in the environment and public health. Parasitological tests performed on 80 samples of affluent and effluent waters from three anaerobic biodigestors with flexible structure from swine farms of Caldas - Colombia, showed the presence of Isospora suis (45%), Eimeria suis (42.5%), E. espinosa (35%), Strongyloides ransomi (28.8%), E. perminuta (12.5%), E. cerdonis (3.8%), and E. porci (2.5%). The additional finding of eggs of Taenia spp. in 10% of the samples was probably caused by a connection between the human sewage system and the biodigester. Although we observed a mean decrease of 65.6% of parasites, these levels were insufficient to meet the minimum requirement set by Engelberg's guidelines regarding water quality. This study demonstrates the serious environmental impact that an inadequately treated animal wastewater represents, and has important implications for water resources and human health.

Resíduos sólidos e líquidos oriundos da suinocultura representam desafios importantes para a produção animal no que diz respeito ao impacto do meio ambiente e em saúde pública. Testes parasitológicos foram realizados em 80 amostras de águas afluentes e efluentes de três biodigestores anaeróbicos com estrutura flexível em granjas de Caldas - Colômbia, demonstrando a presença de Isospora suis (45%), Eimeria suis (42,5%), E. espinosa (35%), Strongyloides ransomi (28,8%), E. perminuta (12,5%), E. cerdonis (3,8%) e E. porci (2,5%). A constatação adicional de ovos de Taenia spp. em 10% das amostras foi provavelmente causada pela conexão entre o sistema de esgoto humano e o biodigestor. Embora fosse observada uma diminuição média de 65,6% dos parasitas, estes níveis são insuficientes para atender ao requisito mínimo estabelecido pelas diretrizes Engelberg em relação à qualidade da água. Este estudo demonstra o grave impacto ambiental que águas residuais animais inadequadamente tratadas representam, bem como as implicações nos recursos hídricos e na saúde humana.

The influence of environmental characteristics on the distribution of ciliates (Protozoa, Ciliophora) in an urban stream of southeast Brazil / Influência das características ambientais sobre distribuição dos ciliados (Protozoa, Ciliophora) em um córrego urbano no sudeste brasileiro

The aim of this research was to study the ciliated protozoa community at three sampling stations that receive different levels of domestic sewage along the São Pedro Stream in the municipality of Juiz de Fora, Minas Gerais, Brazil, in order to determine the influence of organic pollution on this community and to assess the feasibility of using ciliates as water quality indicators. Four physical-chemical parameters of the water samples were evaluated: dissolved oxygen concentration, electrical conductivity, pH and temperature. The sediment was obtained manually, using dredges with capacity of 300 mL, at each collection point. Point 1 was located in a rural region that receives a low sewage load, while Points 2 and 3 were located in populated regions receiving high sewage loads. We found 22 ciliate species, of which 18 are included in the saprobic system and are considered bioindicators. These showed beta-mesosaprobic environments at Point 1 and alfa-mesosaprobic to polisaprobic environments at Points 2 and 3. The low levels of dissolved oxygen and the high electrical conductivity values at Points 2 and 3, together with the strong similarity between the ciliate taxocenoses of these points and the weak similarity between Point 1 and the other two, confirm the high sewage loads received at the latter two points. The combination of the biological indicators and physical-chemical analyses therefore proved itself to be an efficient method of evaluating water quality, and has excellent potential to support decisions on the conservation of headwaters and recuperation of degraded environments in lotic systems.

O objetivo deste trabalho foi estudar a comunidade de protozoários ciliados em três estações amostrais, que recebem diferentes níveis de lançamento de esgoto doméstico, ao longo do córrego São Pedro, Juiz de Fora, MG, a fim de se determinar a influência da poluição orgânica sobre a composição e distribuição desta comunidade e avaliar a viabilidade da utilização dos ciliados como indicadores da qualidade da água. Foram mensurados quatro parâmetros físico-químicos da água amostrada: teor de oxigênio dissolvido, condutividade elétrica, pH e temperatura. Amostras do sedimento foram obtidas manualmente com o auxílio de dragas, com capacidade de 300 mL, em cada ponto de coleta. O ponto 1, localizado em uma região rural, recebe baixa carga de esgoto, enquanto os pontos 2 e 3, localizados em regiões com ampla ocupação humana, recebem altas cargas de esgoto. Foram registradas 22 espécies de ciliados, sendo que 18 estão incluídas no sistema sapróbio e são consideradas bioindicadoras de ambientes beta-mesossapróbio no ponto 1 e de alfa a polissapróbio nos pontos 2 e 3. Os baixos valores de oxigênio dissolvido e os altos valores de condutividade elétrica registrados nas estações 2 e 3, juntamente com a similaridade entre a taxocenose de ciliados destas estações e a baixa similaridade entre a estação 1 e as demais, confirmaram as altas cargas de esgoto recebidas nestas estações. A união do método biológico com a análise físico-química mostrou-se, portanto, um eficiente método na avaliação da qualidade da água, e apresenta grande potencial de utilização em tomadas de decisões relativas à conservação de nascentes e recuperação de ambientes degradados em sistemas lóticos.

Microbiological assessment of sludge treatment at the 6[th] October wastewater treatment plant, Giza, Egypt

This study was carried out to assess the sludge treatment processes applied at the 6[th] October wastewater treatment plant [WWTP], Giza, Egypt. The applied processes included aerobic digestion followed by gravity thickening, filter-pressing and drying beds [in emergency cases]. Bacteriological, virological and parasitological evaluation was carried out. Results showed that aerobic digestion removed two log units of total and faecal coliforms and one log unit of Escherichia coli [E. coli] and feacal streptococci. The removal percentages of coliphage and enteroviruses after aerobic digestion were 97.0 and 58.3%, resectively. The salmonellae disappeared from sludge after aerobic digestion. Qualitative and quantitative decrease in helminth ova [Ascaris, Trichuris, Trichostrongylus, Taenia and Hymenoleois] was observed after aerobic digestion and thickening stages. An average decreae of 4 log units for total and faecal coliforms, 3 log units for each of faecal streptococci and coliphage and 2 log units for E. coli through sludge treatment process was observed. Enteroviruses, helminth ova and enteric protozoa were completely eliminated from sludge after 6 months in drying beds. The loads of faecal coliform, E coli, faecal streptococci and coliphage in the dried sludge were 6.5 x 10[2], 1.6 x10[2], 4.5 x 10[2], 4.56 x 10[2] cfu and 8.5 x 10[1] pfu/g, respectively. Microbiological quality of the final treated sludge complied with the regulations of Untied States Environmental Protection Agency

Emerging pathogen Cyclospora cayetanensis infection in Nepal.

Cylospora cayetanensis, an emerging parasitic pathogen of human is being increasingly recognized throughout the world, however the means of transmission and the possibility of a reservoir host remain an enigma. A longitudinal study on cyclosporiasis in different parts of Nepal was carried out from April, 1995 until November, 2000. Fecal specimens were collected from symptomatic and asymptomatic patients. The data shows a distinct seasonality with the highest infection rates occurring during the summer and rainy season of the year. Attempts have been made to determine the sources of infection and possible reservoir hosts. Stools were examined from nearly 700 animals such as chickens, pigs, buffalos, cows, dogs, cats, monkeys, rats, mice and pigeons. In addition, vegetable farms around the Kathmandu Valley were examined during the seasonal high and low periods of transmission. C. cayetanensis-like oocysts were found in sewage water and from vegetable washings on five occasions during June, July, August, October, and November. Similarly, C. cayetanensis-like oocysts were recovered from mice, rats, chickens, and dogs. These results suggest that these sources may be important in the transmission of this parasitosis. However, further studies will be required to obtain definitive answers on transmission.

Study of Cyclospora cayetanensis in health care facilities, sewage water and green leafy vegetables in Nepal.

Cyclospora cayetanensis, a newly emerging parasite, is endemic in Nepal. A total of 2,123 stool specimens were collected from 3 health care facilities based on clinical symptoms during the period between 1995 to October, 1998. Out of these specimens, cayetanensis oocysts were found in 632 (29.8%). To identify possible sources for Cyclospora infection, drinking water, sewage water, green-leafy vegetables including fecal samples of various animals were collected and examined. The vegetable leaves were washed in distilled water then the washings, sewage water and drinking water were centrifuged and the sediment were examined microscopically. As a result, oocyst of Cyclospora were identified in sewage water and vegetable washings on four different occasions in June, August, October and November. The positive results were also confirmed as C. cayetanensis by development of 2 sporocysts after 2 week incubation period in potassium dichromate. A survey of 196 domestic animals from the same areas demonstrated that two chickens were positive for Cyclospora-like organism and others were negative. Although further studies are needed to clarify the direct link between Cyclospora infection and these sources, the results suggest that sewage water, green leafy vegetables are possible sources of infection and chickens could be possible reservoir host of Cyclospora in Nepal.

Helminthic infections associated with the use of raw wastewater for agricultural purposes in Beni Mellal, Morocco

We aimed to determine the possible risks associated with raw wastewater use for agricultural purposes, particularly the transmission of helminthic infections among children. In a randomly-selected sample of 1343 children, 740 of them were from five regions using raw wastewater for agriculture, while 603 were from four control regions that do not use such water. The prevalence of helminthic infection was found to be significantly higher among children in regions using wastewater, compared to control regions. Behavioural characteristics and public water supply were found to be associated with higher infection rates. Adequate treatment of wastewater as well as public health education are highly recommended