Caracterización de pacientes con enfermedades asociadas a inmunosupresión con infecciones por Cryptosporidium spp. atendidos en un hospital pediátrico referencial de Argentina, 2018-2023 / Description of the characteristics of patients with diseases associated with immunosuppression and Cryptosporidium spp. infection seen at a referral children's hospital in Argentina, 2018­2023

Cryptosporidium spp. es un protozoario productor de diarrea. Los pacientes inmunocomprometidos pueden desarrollar formas clínicas graves y persistentes. Se describen las características de pacientes con enfermedad de base asociada a inmunosupresión (EAI) con infección por Cryptosporidium spp. (IC) atendidos en un hospital pediátrico referencial de Argentina entre los años 2018 y 2023. Se analizaron datos demográficos, EAI, características de la diarrea y coinfecciones. Se incluyeron 30 pacientes con EAI e IC. La mayoría registró trasplante de órgano sólido, neoplasia hematológica e inmunodeficiencia primaria. Dieciocho presentaron diarrea persistente al momento del diagnóstico. Seis pacientes registraron coinfecciones. Se debe considerar la criptosporidiosis en el diagnóstico diferencial de enfermedad diarreica aguda o persistente en niños con distintos tipos de EAI, como el trasplante de órgano sólido, neoplasias hematológicas e inmunodeficiencias primarias.

Cryptosporidium spp. is a diarrhea-causing protozoan. Immunocompromised patients may develop severe and persistent clinical forms. Here we describe the characteristics of patients with an underlying disease associated with immunosuppression (DAI) and Cryptosporidium spp. infection seen at a referral children's hospital in Argentina between 2018 and 2023. Demographic data, DAI, diarrhea characteristics, and co-infections were analyzed. A total of 30 patients with DAI and cryptosporidiosis were included. Most of them had undergone a solid organ transplant, had a hematologic neoplasm, or primary immunodeficiency. Persistent diarrhea was observed in 18 patients at the time of diagnosis. Co-infections were recorded in 6 patients. Cryptosporidiosis should be considered in the differential diagnosis of acute or persistent diarrhea in children with different types of DAI, such as solid organ transplant, hematologic neoplasms, and primary immunodeficiencies.

Diversity of gastrointestinal parasites and molecular characterization of Giardia duodenalis in free-living and captive howler monkeys (Alouatta guariba clamitans) in southern Brazil.

Non-human primates (NHPs) are the group that most share infectious agents with humans due to their close taxonomic relationship. The southern brown howler monkeys (Alouatta guariba clamitans) are endemic primates from Brazil and Argentina's Atlantic Forest. This study aimed to investigate the presence of intestinal parasites in free-living (FL) and captive (CA) southern brown howler monkeys. Thirty-nine stool samples were collected in two areas in southern Brazil, 15 FL and 24 CA. Stool sediments obtained by centrifugal sedimentation technique were used for microscopic analysis and direct immunofluorescence assay and evaluated by molecular analysis through amplification and sequencing of TPI fragments. Intestinal parasites Giardia duodenalis, Cryptosporidium spp., and Trypanoxyuris minutus were detected at coproparasitological analysis. This is the first report of the presence of Cryptosporidium spp. in free-living howlers. The molecular characterization of G. duodenalis isolates indicated assemblage B for the first time found in free-living A. guariba clamitans. The high prevalence of G. duodenalis transmission in CA howler monkeys can be explained by direct contact with humans and frequent soil contact. The presence of a potentially zoonotic assemblage in these animals indicates that the process of fragmentation and cohabitation with humans and livestock affects the wildlife, thus indicating a need for eco-health measures.

Estimation and evaluation of the risks of protozoa infections associated to the water from a treatment plant in southern Brazil using the Quantitative Microbiological Risk Assessment Methodology (QMRA).

In this study, the Quantitative Microbial Risk Assessment (QMRA) methodology was applied to estimate the annual risk of Giardia and Cryptosporidium infection associated with a water treatment plant in southern Brazil. The efficiency of the treatment plant in removing protozoa and the effectiveness of the Brazilian legislation on microbiological protection were evaluated, emphasizing the relevance of implementing the QMRA in this context. Two distinct approaches were employed to estimate the mechanical removal of protozoa: The definitions provided by the United States Environmental Protection Agency (USEPA), and the model proposed by Neminski and Ongerth. Although the raw water collected had a higher concentration of Giardia cysts than Cryptosporidium oocysts, the estimated values for the annual risk of infection were significantly higher for Cryptosporidium than for Giardia. From a general perspective, the risk values of protozoa infection were either below or very near the limit set by the World Health Organization (WHO). In contrast, all the risk values of Cryptosporidium infection exceeded the threshold established by the USEPA. Ultimately, it was concluded that the implementation of the QMRA methodology should be considered by the Brazilian authorities, as the requirements and guidelines provided by the Brazilian legislation proved to be insufficient to guarantee the microbiological safety of drinking water. In this context, the QMRA application can effectively contribute to the prevention and investigation of outbreaks of waterborne disease.

Description of the characteristics of patients with diseases associated with immunosuppression and Cryptosporidium spp. infection seen at a referral children's hospital in Argentina, 2018-2023. / Caracterización de pacientes con enfermedades asociadas a inmunosupresión con infecciones por Cryptosporidium spp. atendidos en un hospital pediátrico referencial de Argentina, 2018-2023.

Cryptosporidium spp. is a diarrhea-causing protozoan. Immunocompromised patients may develop severe and persistent clinical forms. Here we describe the characteristics of patients with an underlying disease associated with immunosuppression (DAI) and Cryptosporidium spp. infection seen at a referral children's hospital in Argentina between 2018 and 2023. Demographic data, DAI, diarrhea characteristics, and co-infections were analyzed. A total of 30 patients with DAI and cryptosporidiosis were included. Most of them had undergone a solid organ transplant, had a hematologic neoplasm, or primary immunodeficiency. Persistent diarrhea was observed in 18 patients at the time of diagnosis. Co-infections were recorded in 6 patients. Cryptosporidiosis should be considered in the differential diagnosis of acute or persistent diarrhea in children with different types of DAI, such as solid organ transplant, hematologic neoplasms, and primary immunodeficiencies.

Cryptosporidium spp. es un protozoario productor de diarrea. Los pacientes inmunocomprometidos pueden desarrollar formas clínicas graves y persistentes. Se describen las características de pacientes con enfermedad de base asociada a inmunosupresión (EAI) con infección por Cryptosporidium spp. (IC) atendidos en un hospital pediátrico referencial de Argentina entre los años 2018 y 2023. Se analizaron datos demográficos, EAI, características de la diarrea y coinfecciones. Se incluyeron 30 pacientes con EAI e IC. La mayoría registró trasplante de órgano sólido, neoplasia hematológica e inmunodeficiencia primaria. Dieciocho presentaron diarrea persistente al momento del diagnóstico. Seis pacientes registraron coinfecciones. Se debe considerar la criptosporidiosis en el diagnóstico diferencial de enfermedad diarreica aguda o persistente en niños con distintos tipos de EAI, como el trasplante de órgano sólido, neoplasias hematológicas e inmunodeficiencias primarias.

Intestinal protozoa in returning travellers: a GeoSentinel analysis from 2007 to 2019.

BACKGROUND: Prolonged diarrhoea is common amongst returning travellers and is often caused by intestinal protozoa. However, the epidemiology of travel-associated illness caused by protozoal pathogens is not well described. METHODS: We analysed records of returning international travellers with illness caused by Giardia duodenalis, Cryptosporidium spp., Cyclospora cayetanensis or Cystoisospora belli, reported to the GeoSentinel Network during January 2007-December 2019. We excluded records of travellers migrating, with an unascertainable exposure country, or from GeoSentinel sites that were not located in high-income countries. RESULTS: There were 2517 cases, 82.3% giardiasis (n = 2072), 11.4% cryptosporidiosis (n = 287), 6.0% cyclosporiasis (n = 150) and 0.3% cystoisosporiasis (n = 8). Overall, most travellers were tourists (64.4%) on long trips (median durations: 18-30 days). Cryptosporidiosis more frequently affected people < 18 years (13.9%) and cyclosporiasis affected people ≥ 40 years (59.4%). Giardiasis was most frequently acquired in South Central Asia (45.8%) and sub-Saharan Africa (22.6%), cryptosporidiosis in sub-Saharan Africa (24.7%) and South-Central Asia (19.5%), cyclosporiasis in South East Asia (31.3%) and Central America (27.3%), and cystoisosporiasis in sub-Saharan Africa (62.5%). Cyclosporiasis cases were reported from countries of uncertain endemicity (e.g. Cambodia) or in countries with no previous evidence of this parasite (e.g. French Guiana). The time from symptom onset to presentation at a GeoSentinel site was the longest amongst travellers with giardiasis (median: 30 days). Over 14% of travellers with cryptosporidiosis were hospitalized. CONCLUSIONS: This analysis provides new insights into the epidemiology and clinical significance of four intestinal protozoa that can cause morbidity in international travellers. These data might help optimize pretravel advice and post-travel management of patients with travel-associated prolonged gastrointestinal illnesses. This analysis reinforces the importance of international travel-related surveillance to identify sentinel cases and areas where protozoal infections might be undetected or underreported.

Molecular characterization of Cryptosporidium in ruminants and observation of natural infection by Cryptosporidium andersoni in sheep from Paraná, Brazil.

The aim of this study was to identify Cryptosporidium species found in cattle and sheep in Paraná, southern region of Brazil. Individual fecal samples from 458 bovines and 101 sheep were submitted for molecular analysis by PCR and nested PCR using specific primers for sequences of the 18S ribosomal unit (rRNA). Positive samples were analyzed using restriction fragment length polymorphism (RFLP), followed by genetic sequencing for species confirmation. The occurrence of Cryptosporidium was 11.27% (63/559). The highest occurrence was detected in lambs (12/59, 20.33%). From the 63 positive samples, it was possible to identify the species in 58 of them by RFLP and genetic sequencing. Five species of Cryptosporidium were identified: Cryptosporidium andersoni, Cryptosporidium bovis, Cryptosporidium ryanae, Cryptosporidium xiaoi, and Cryptosporidium parvum. The most prevalent species was C. andersoni (41.38%) and the least predominant was C. parvum (10.34%). The most abundant species of Cryptosporidium in dairy calves were C. andersoni (11/25) and C. ryanae (6/25). Of the 17 positive sheep, nine (52.94%) were infected with C. andersoni. This finding is the first report on the occurrence of C. andersoni in naturally infected sheep in Brazil and the first observation of a high absolute occurrence of this Cryptosporidium species in sheep.

Occurrence of gastrointestinal parasites in dogs in a rural area of Santa Catarina, Brazil.

We investigated the occurrence of gastrointestinal parasites in dogs in rural areas of the municipality of Painel, Santa Catarina, Brazil. For this, 91 canine feces samples were collected from 82 farms between August, 2017 and January, 2018. These fecal materials were processed using the techniques of spontaneous sedimentation, centrifugation-floatation in zinc sulfate and Ziehl-Neelsen staining. A questionnaire in the form of an interview was administered to the dogs' owner and information about the farm and the main care provided for the dogs was obtained. Among 91 sampled dogs, 27 (29.7%) were positive for at least one parasite species. Ancylostoma was the most prevalent genus (16.5%), followed by Giardia duodenalis (14.3%), Trichuris vulpis (6.6%), Toxocara canis (5.5%), Entamoeba spp. (4.8%), Cryptosporidium spp. (3.3%) and Taeniidae (1.1%). Most dogs do not receive veterinarian care and rarely received antiparasitic treatment. They were free to roam and had free access to animal remains and garbage, which was reflected in the significant associations with the occurrence of parasites that were found. We conclude that rural dogs harbor gastrointestinal parasites, but that their owners are unaware of the risks that these parasites can bring to human health.

Cryptosporidium infection and associated risk factors among children from rural areas in Peru: a nested case-control study.

This study aimed to detect the frequency of Cryptosporidium infection and associated risk factors among children from rural areas in Peru. A case-control study was conducted, nested in a cohort in two rural communities that included children between 6 and 13 months who were followed for 6 months. Cases were children whose fecal samples tested positive for Cryptosporidium infection using an immunochromatography test. Logistic regression was used to calculate odds ratios (ORs) and 95% confidence intervals (CIs) to analyze risk factors associated with Cryptosporidium infection. Among 72 children, 13 (18%) were cases. Cryptosporidium infection was associated with below secondary education of the mother (OR 7.62, 95% CI 1.50-36.72) and with having more siblings living at home (OR 1.71, 95% CI 1.04-2.82). An important frequency of Cryptosporidium infection among children from rural areas in Peru was reported, more research is needed to understand its true burden and risk factors among children in Peru.

Zoonotic Cryptosporidium meleagridis in urban invasive monk parakeets.

Zoonotic Cryptosporidium meleagridis was identified in invasive monk parakeets (Myiopsitta monachus) from Santiago metropolis in Chile. Oocysts were isolated from nestlings' faeces, and a fragment of 18S ribosomal RNA gene was amplified and sequenced. This finding emphasizes the importance of permanent pathogen monitoring in synanthropic species with wide global distribution.

Prevalence of Cryptosporidium spp. in Horses from communities of the Mapuche native people, Araucanía Region, Chile.

BACKGROUND: There are no studies of potential zoonotic diseases in Mapuche communities' horses. OBJECTIVES: The objective of this study was to determine the prevalence of Cryptosporidium in horses of the Mapuche communities. STUDY DESIGN: This was a cross-sectional study. METHODS: Faecal samples from 100 randomly selected horses (n = 100) were taken from rural Mapuche communities from four municipalities from the Araucanía Region. These samples were processed with the modified Ziehl-Neelsen staining technique and grouped by sex, age and municipality. RESULTS: The general prevalence of Cryptosporidium spp. was 67.0% (n = 67). The prevalence was 51.0% (n = 51) in males and 49.0% (n = 49) in females, and there is no gender association to the presentation of Cryptosporidium spp. The prevalence by municipality was 60.0%, 80.0%, 64.0% and 64.0% in Curarrehue, Lonquimay, Padre las Casas and Teodoro Schmidt, respectively. The above shows no significant association between the sector and the presence of Cryptosporidium spp. The prevalence by age was 95.4% of horses tested positive for Cryptosporidium between birth and 6 years of age. About 27.3% of horses were tested positive in the age group between 7 and 10 years. There was no presence of Cryptosporidium spp. in the age group older than 11 years, showing a significant relationship between the age of the animal and the presence of Cryptosporidium (P Ë‚ 0.05). MAIN LIMITATIONS: The sample analysis did not specifically identify the type of Cryptosporidium, and it was not possible to evaluate the zoonotic risk in the Mapuche communities. CONCLUSIONS: Cryptosporidium spp. is present in working horses in Mapuche communities, with a 67.7% general prevalence, and there is a significant association between this parasite and the age of the horses, being higher in the age group between 0 and 6 years, with a prevalence of 95.4%. There may be a potential zoonotic risk in the Mapuche communities.

HISTORIAL: No hay estudios sobre enfermedades potencialmente zoonoticas en caballos de las comunidades Mapuche. OBJETIVOS: El objetivo de este estudio es determinar la prevalencia de Cryptosporidium en caballos de las comunidades Mapuche. DISEÑO DEL ESTUDIO: Estudio transversal. MÉTODOS: Muestras fecales de cien caballos (n = 100) seleccionados al azar, fueron recolectadas en comunidades Mapuche de cuatro municipalidades de la Región de la Araucanía. Estas muestras fueron procesadas con la técnica de tinción Ziehl-Neelzen modificada y agrupadas por sexo, edad y municipalidad. RESULTADOS: La prevalencia general de Cryptosporidium spp fue de 67.0% (n = 67). La prevalencia fue 51.0% (n = 51) en machos y 49.0% (n = 49) en hembras, y no hay asociación de genero con la presentación de Cryptosporidium spp. La prevalencia por municipalidad fue de 60.0%, 80.0%, 64.0%, 64.0% en Curarrehue, Lonquimay, Padre las Casas y Teodoro Schmidt respectivamente. Lo presentado arriba muestra ninguna asociación significativa entre el sector y la presencia de Cryptosporidium spp. La prevalencia por edad fue de 95.4% en caballos que resultaron ser positivos a Cryptosporidium entre nacimiento y seis años de edad. 27.3% resultaron ser positivos entre siete y diez años. No hubo presencia de Cryptosporidium spp en el grupo mas grande de caballos mayores a once años, mostrando una relación significativa entre la edad del animal y la presencia de Cryptosporidium (P < 0.05). LIMITACIONES PRINCIPALES: El análisis de muestras no fue especifico para poder identificar el tipo de Cryptosporidium, y no fue posible evaluar el riesgo zoonotico en la comunidades Mapuche. CONCLUSIONES: Los resultados de este estudio concluyeron que existe la presencia de Cryptosporidium spp en caballos de trabajo en las comunidades Mapuche; con un 67.0% de prevalencia, y que hay una asociación significativa entre este parasito y la edad de los caballos, siendo mayor en el grupo etario entre 0 y 6 años, con una prevalencia de 95.4%. Estos resultados pueden inferir que podría haber un riesgo zoonotico potencial en las comunidades Mapuche.

Cryptosporidium spp. in dogs - prevalence and genotype distribution

Background: Cryptosporidium spp. is a zoonotic protozoan parasite that affects the gastrointestinal tract of humans and animals. The disease can cause acute and chronic diarrhoea and even death in both humans and animals. In this study, it was aimed to determine the prevalence and genotype distribution of Cryptosporidiosis in shelter dogs in Diyarbakir province located in the Southeastern Anatolia Region of Turkey. Materials, Methods & Results: The animal material of the study consisted of 100 dogs of different breeds and sexes. Faecal samples were collected from the rectum with disposable latex gloves and placed in individual sample containers. All of the samples were examined for Cryptosporidium spp. by Kinyoun Acid Fast and Nested PCR methods. In the Kinyoun Acid Fast staining method, firstly, smear preparations were prepared from fresh faecal samples, fixed in pure methanol for 1 min and allowed to dry. The slides were kept in Kinyoun Carbol-Fuxin for 5 min, dipped in 50% ethyl alcohol, shaken, washed in tap water, kept in 1% sulphuric acid for 2 min and washed in tap water. The slides were kept in methylene blue for 1 min, washed in tap water and allowed to dry. After drying, immersion oil was dripped and examined under a microscope at 100 magnification. DNA extraction was performed from all samples using GeneMATRIX Stool DNA Purification Kit according to the manufacturer's protocol. After Nested PCR analysis was performed. In the PCR step, primers 5'-TTCTAGAGCTAATACATGCG-3' and 5'- CCCATTTCCTTCCTTCGAAACAGGA-3' were used to amplify the 1325 bp gene region. In the nested PCR step, primers 5'- GGAAGGGTTGTATTTATTTATTAGATAAAG-3' and 5'-AAGGAGTAAGGAACAACCTCCA-3' were used to amplify the 826-864 bp gene region. As a result of both methods, a prevalence of 3% was determined. The infection rate was higher in males (3.57%) than females (2.27%) and in younger than 1 year (5.56%) than in older than 1 year (1.56%). The DNA sequences obtained from the sequence analysis of 3 positive PCR samples were analysed in BioEdit software. A phylogenetic tree was constructed with the data set created by using the 18s rRNA gene sequences obtained from the NCBI genbank database and the DNA sequences obtained as a result of the study, and it was shown which Cryptosporidium species the study samples were related to. Today, many Cryptosporidium species have been identified and most of these species have host adaptation. Although C. canis is the most common species in dogs, C. muris, C. meleagridis, and C. parvum have also been detected. Among these species, C. parvum is recognized as a zoonotic species infecting a wide range of mammals. In this study, DNA sequencing of nested PCR positive samples revealed that 3 samples were zoonotic C. parvum. Discussion: This suggests that dogs may be a reservoir for zoonotic transmission of Cryptosporidium. Consequently, it is recommended that people should be informed about the potential for transmission of this protozoan to humans and animals and that control programmes should be implemented, including the prevention of free entry of stray dogs into public places and homes.

Cross-genera amplification and identification of Colpodella sp. with Cryptosporidium primers in fecal samples of zoo felids from northeast China / Amplificação cruzada de gêneros e identificação de Colpodella sp. com iniciadores de Cryptosporidium em amostras fecais de zoofilia do nordeste da China

Abstract The protozoans include many intracellular human pathogens. Accurate detection of these pathogens is necessary to treat the diseases. In clinical epidemiology, molecular identification of protozoan is considered a more reliable and rapid method for identification than microscopy. Among these protozoans, Cryptosporidium considered being one of the important water-borne zoonotic pathogens and a major cause of a diarrheal disease named cryptosporidiosis in humans, domestic animals, and wild animals. This study was aimed to identify Cryptosporidium in zoo felids (N= 56) belonging to different zoo of China, but accidentlly Colpodella was encountered in the zoo felids sample and phylogenetic data confirmed this unexpected amplification from fecal samples using two-step nested-PCR. Phylogenetic analysis revealed the fact about the specific primers used previously by many researchers and cross-genera amplification. We came to know that genetically sequenced amplicon gives more accurate identification of species. This study suggests more investigation on Colpodella which has been neglected previously but gains the attention of researchers after identified from humans and animals and has been known to correlate with neurological symptoms in patients.

Resumo Os protozoários incluem muitos patógenos humanos intracelulares. A detecção acurada desses patógenos é necessária para tratar as doenças. Na epidemiologia clínica, a identificação molecular de protozoários é considerada o método de identificação mais confiável e rápido do que a microscopia. Entre esses protozoários, o Cryptosporidium é considerado um dos importantes patógenos zoonóticos transmitidos pela água e uma das principais causas de uma doença diarreica denominada criptosporidiose em humanos, animais domésticos e selvagens. Este estudo teve como objetivo identificar Cryptosporidium em zoofelídeos (N = 56) pertencentes a diferentes zoológicos da China, mas acidentalmente Colpodella foi encontrada na amostra de zoofelídeos e os dados filogenéticos confirmaram essa amplificação inesperada de amostras fecais usando nested-PCR em duas etapas. A análise filogenética revelou o fato sobre os primers específicos usados ​​anteriormente por muitos pesquisadores e a amplificação entre gêneros. Ficamos sabendo que o amplicon sequenciado geneticamente fornece uma identificação mais acurada das espécies. Este estudo sugere mais investigação sobre Colpodella, que foi negligenciada anteriormente, mas ganha a atenção dos pesquisadores depois de identificada em humanos e animais e é conhecida por se correlacionar com sintomas neurológicos em pacientes.

Occurrence and risk factors of bovine cryptosporidiosis in Brazil ­ Systematic Review / Ocorrência de criptosporidiose bovina no Brasil ­ Revisão sistemática

Protozoa of the genus Cryptosporidium are parasites capable of causing animal rearing losses due to diarrhea outbreaks. This review aimed to determine the prevalence of and potential risk factors for bovine cryptosporidiosis in Brazil. The search was carried out in the databases Science Direct, Scopus, Web of Science, and SciELO. The search strategy used the descriptors "Cryptosporidium," "cryptosporidiosis," "cattle," "calves," and "Brazil." Nine studies conducted in different Brazilian states between 2008 and 2019 were selected. The age of the evaluated animals ranged from 0 to >36 months. The main risk factors were the low age of the animals, collective pens, presence of diarrhea, contact with other animals, and dairy breed. The average prevalence rate was 30.51%. The species identified were C. ryanae, C. parvum, and C. bovis. Optical microscopy was used as a diagnostic method for oocysts in fecal samples and to determine the species, and PCR was used in three studies. The average prevalence of Cryptosporidium spp. in cattle in Brazil was 30.51%, ranging from 10.2 to 62.5%. The main risk factors found were young animals, the presence of animals with diarrhea, contact with other animals, dairy breeds, and collective pens.(AU)

Protozoários do gênero Cryptosporidium são parasitas capazes de causar perdas durante a criação animal devido à surtos de diarreia. Esta revisão foi realizada para determinar a prevalência e os potenciais fatores de risco para a ocorrência de criptosporidiose bovina no Brasil. A busca foi conduzida nas bases de dados Science Direct, Scopus, Web of Science e Scielo. A estratégia de busca usou os descritores "Cryptosporidium", "cryptosporidiosis", "cattle", "calves" e "Brazil". Nove estudos conduzidos em diferentes estados do Brasil foram selecionados, com ocorrências entre 2008 e 2019. A idade dos animais avaliados variou de 0 acima de 36 meses. Os principais fatores de risco determinados foram animais jovens, baias coletivas, presença de animais com diarreia, contato com outros animais e raça leiteira. A taxa de prevalência média foi de 30,51%. As espécies identificadas foram C. ryanae, C parvum e C. bovis. Microscopia óptica foi usada como método de diagnóstico de oocistos nas amostras de fezes e, para determinação das espécies, a técnica PCR foi usada em três estudos. Concluiu-se que a prevalência média do protozoário Cryptosporidium spp. em bovinos no Brasil foi 30,51%, variando de 10,2 a 62,5%. Os principais fatores de risco foram animais jovens, presença de animais com diarreia, contato com outros animais, baias coletivas e raças leiteiras.(AU)

Prevalence of human cryptosporidiosis in the Americas: systematic review and meta-analysis.

Cryptosporidiosis is a disease caused by the Cryptosporidium spp parasite. As some species of Cryptosporidium have a wide host spectrum, the characterization of the pathogen at the species or genotype level is of great importance to define the sources of infection for humans and the potential for public health. This study investigated the diversity of the genus Cryptosporidium spp. in humans from all over the American continent and observed whether the method used to search for the parasite influenced the prevalence found in the Americas. This systematic review was carried out using the Pubmed, Science direct, Lilacs, Scielo, and Scopus databases with publications from January 1, 2010, to December 31, 2020. For data synthesis, the PRISMA flowchart was used and for the meta-analysis we used the MetaXL program. Of the selected publications, 57, 9 and 16 belonged to the region of South, Central and North America, respectively. The prevalence found for South, Central, and North America was 7%, 7%, and 8%, respectively, when analyzing publications that used only the microscopy method. When we analyzed the publications that used immunological and molecular methods, we found prevalences of 10%, 9%, and 21% for South, Central, and North America, respectively. The C. hominis subtype IbA10G2 was the most reported in the American continent, followed by subtype IeA11G3T3 and, for C. parvum, subtype IIaA15G2RI was the most reported. In conclusion, Cryptosporidium spp. is present throughout the American continent and its prevalence is higher when immunological and/or molecular methods are used, in addition to direct microscopic examination.

High occurrence of viable forms of Cryptosporidium and Giardia in domestic sewage from an agricultural region of Brazil.

Cryptosporidium and Giardia are the main etiologies of waterborne outbreaks caused by protozoa. These parasites are commonly detected in wastewater; however, there is little knowledge about the concentration of viable forms in treated sewage, mainly in small communities. To understand more about the presence of viable oocysts and cysts in domestic sewage, we monitored the affluent and effluent of a wastewater treatment plant (WWTP) in inner-city Brazil. Ten samplings and seven follow-ups were performed in 2020. Samples were concentrated by centrifugation, filtration and purified by fluctuation. Viability was accessed by propidium-monoazide (PMA) associated with nPCR and qPCR. Both viable protozoa were detected in all raw sewage samples (average: 438.5 viable oocysts/L). Regarding treated sewage, Cryptosporidium was detected in all of the samples (average: 92.8 viable oocysts/L) and Giardia was detected in 70% with viable cysts in 30%. Considering the follow-ups, 31.17% of Cryptosporidium viable oocysts remained in the effluent after the treatment. High amounts of Cryptosporidium and a high frequency of Giardia were detected, therefore both arrived at WWTP and were discharged into the river. These alert the presence of agro-industrial effluents into domestic sewage and demonstrated the effectiveness of the concentration technique for monitoring protozoa in wastewater.

Cryptosporidium spp. and Giardia spp. in wild rodents: using occupancy models to estimate drivers of occurrence and prevalence in native forest and exotic Pinus radiata plantations from Central Chile.

The conversion of native forest to forestry plantations is a worldwide practice, affecting biodiversity and host-parasite interactions. One of the most common timber plantations in the world are monocultures of Monterey pine (Pinus radiata). Using occupancy models, we analyzed the occurrence and prevalence of Cryptosporidium spp. oocysts and Giardia spp. cysts in fecal samples of wild rodents from a landscape dominated by extensive Monterey pine plantations in central Chile. We aimed to assess drivers of parasite infection such as habitat type, abundance of rodent hosts, species richness, and season. Small mammals were sampled seasonally for two years in three habitat types: native forests, adult pine plantations and young pine plantations. A total of 1091 fecal samples from seven small mammal species were analyzed by coprological analysis. Occurrence probability of Cryptosporidium spp. and Giardia spp. was similar for the most abundant rodent species (Abrothrix longipilis, A. olivacea, and Oligoryzomys longicaudatus) and for all habitat types. For Cryptosporidium spp., variation in prevalence was mostly explained by season with higher prevalence during winter season and lower during spring. For Giardia spp., the prevalence was significantly higher in young pine plantations, followed by adult pine plantations and native forests. In addition, higher prevalence of Giardia spp. was associated with lower host richness, suggesting a possible dilution effect. Our findings reveal that Monterey pine plantations increase Giardia spp. transmission among rodents, but has no clear effect on Cryptosporidium spp., providing evidence that the impact of land use on parasitism can be idiosyncratic. Since both parasites have zoonotic potential, our findings may be useful for land use planning and management considering health issues.

Assessment microbial risks for the presence of Cryptosporidium spp. and Giardia spp. based on the surveillance of the water supply systems in Colombia, 2014-2018.

Cryptosporidium spp. and Giardia spp. cause gastrointestinal diseases of zoonotic origin as well transmitted from person to person, being various reported outbreaks associated with water. The infecting (oo)cyst forms of these parasites are highly resistant to water treatments such as chlorine disinfection and fast filtration. The objective of this study was to assess the microbial risk of infection and symptomatic illness by the ingestion of Giardia spp. and Cryptosporidium spp. in water for human consumption in Colombia, based on the results of water quality surveillance. The detection method was according to the USEPA method 1623. Concentration data of the different points of distribution were grouped according to the pathogen and type of treatment (no treatment; chlorine treatment; chlorine treatment + coagulant). Annual microbial risks of infection and symptomatic diseases were estimated using the quantitative microbial risk assessment approach that included parasite concentrations, the dose-response model, the ingestion rates of water by children and adults, and the morbidity rate of the diseases. The mean annual microbial risk of infection for Giardia spp. was 29.8% for treated water and 50.4% for untreated water, while being 6.0% and 17.7%, respectively, for Cryptosporidium spp. Microbial risk of symptomatic illness for Giardia spp, was 8.2% for treated water and 13.9% for untreated water, while being 3.6% and 10.6%, respectively, for Cryptosporidium spp. The estimated annual microbial risks of infection exceeded the acceptable value of 10-4 (0.01%) recommended by USEPA. Results obtained in this study suggest the need to reduce the microbial risk of infection to protozoan parasites by improving the water treatment, by adopting better handling practices for livestock manure and treatment processes of human feces. PRACTITIONER POINTS: The presence of Cryptosporidium spp was identified in 28 (6.2%) samples and Giardia spp in 29 (6.4%) in water for human consumption in Colombia. The mean annual risk of symptomatic illness due to infection by Giardia spp or Cryptosporidium spp ranges from 33.6%, for treated water, to 58.1%, for untreated water. Annual risks ingestion of protozoa studying in water for human exceed of 10-4 (0.01%) recommended by USEPA.

Multilocus Sequence Typing helps understand the genetic diversity of Cryptosporidium hominis and Cryptosporidium parvum isolated from Colombian patients.

Multilocus Sequence Typing has become a useful tool for the study of the genetic diversity and population structure of different organisms. In this study, a MLST approach with seven loci (CP47, MS5, MS9, MSC6-7, TP14, and gp60) was used to analyze the genetic diversity of Cryptosporidium hominis and Cryptosporidium parvum isolated from 28 Colombian patients. Five Cryptosporidium species were identified: C. hominis, C. parvum, Cryptosporidium felis, Cryptosporidium meleagridis, and Cryptosporidium suis. Unilocus gp60 analysis identified four allelic families for C. hominis (Ia, Ib, Id, and Ie) and two for C. parvum (IIa and IIc). There was polymorphic behavior of all markers evaluated for both C. hominis and C. parvum, particularly with the CP47, MS5, and gp60 markers. Phylogenetic analysis with consensus sequences (CS) of the markers showed a taxonomic agreement with the results obtained with the 18S rRNA and gp60 gene. Additionally, two monophyletic clades that clustered the species C. hominis and C. parvum were detected, with a higher number of subclades within the monophyletic groups compared to those with the gp60 gene. Thirteen MLG were identified for C. hominis and eight for C. parvum. Haplotypic and nucleotide diversity were detected, but only the latter was affected by the gp60 exclusion from the CS analysis. The gene fixation index showed an evolutionary closeness between the C. hominis samples and a less evolutionary closeness and greater sequence divergence in the C. parvum samples. Data obtained in this work support the implementation of MLST analysis in the study of the genetic diversity of Cryptosporidium, considering the more detailed information that it provides, which may explain some genetic events that with an unilocus approach could not be established. This is the first multilocus analysis of the intra-specific variability of Cryptosporidium from humans in South America.

Evaluación de pruebas inmunológicas en el diagnóstico de Giardia duodenalis y Cryptosporidium spp, Honduras / Evaluation of immunologic tests in the diagnosis of Giardia duodenalis and Cryptosporidium spp., Honduras

Antecedentes: No conocemos datos sobre evaluación de pruebas inmunológicas para mejorar el diagnóstico de Giardia duodenalis y Cryptosporidium spp., agentes etiológicos de diarrea de importancia mundial, en Honduras. Objetivos: Comparar dos pruebas inmunológicas para el diagnóstico de Giardia y Cryptosporidium spp. con microscopía de rutina y determinar su aplicabilidad local. Métodos: Estudio descriptivo transversal. En 2013, 134 muestras de heces recibidas en el Servicio de Parasitología del Hospital Escuela (HE) y 67 muestras del Centro de Salud Alonso Suazo (CSAS) se analizaron con una Prueba Rápida Inmunocromatográfica (PDR). En 2019-2020, 60 muestras de heces del HE se analizaron con una prueba inmunoenzimática ELISA. El protocolo de rutina incluyó examen directo en solución salina y solución de Lugol, coloración tricrómica y coloración ácido resistente modificada (ARM) (HE) y examen directo en solución salina y solución de Lugol (CSAS). Resultados: Cada prueba inmunológica mostró mayor positividad que la microscopía: en 134 muestras del HE para Giardia (6.7% vs 4.5%) y Cryptosporidium (3.7% vs 0.7%), similar en 67 muestras del CSAS (14.9% vs 7.5% para Giardia; 0.7% para Cryptosporidium con la prueba inmunológica). De 60 muestras analizadas por ELISA en HE, 31.7% fue positiva por Giardia vs 18.3% en examen directo y 23.3% en coloración tricrómica; 6.7% positiva por Cryptosporidium spp. vs 3.3% por coloración ARM. Discusión: Pruebas inmunológicas aumentaron significativamente el diagnóstico de ambas parasitosis; sin embargo, publicaciones sobre pruebas similares ofrecieron resultados no concluyentes. Por costo elevado podrían reservarse para pacientes pediátricos, pacientes inmunocomprometidos en hospitales, complementando microscopía. Los laboratorios de salud deben fortalecer capacidad diagnóstica...(AU)

An Overview of Mucosa-Associated Protozoa: Challenges in Chemotherapy and Future Perspectives.

Parasitic infections caused by protozoans that infect the mucosal surfaces are widely neglected worldwide. Collectively, Entamoeba histolytica, Giardia lamblia, Cryptosporidium spp. and Trichomonas vaginalis infect more than a billion people in the world, being a public health problem mainly in developing countries. However, the exact incidence and prevalence data depend on the population examined. These parasites ultimately cause pathologies that culminate in liver abscesses, malabsorption syndrome, vaginitis, and urethritis, respectively. Despite this, the antimicrobial agents currently used to treat these diseases are limited and often associated with adverse side effects and refractory cases due to the development of resistant parasites. The paucity of drug treatments, absence of vaccines and increasing problems of drug resistance are major concerns for their control and eradication. Herein, potential candidates are reviewed with the overall aim of determining the knowledge gaps and suggest future perspectives for research. This review focuses on this public health problem and focuses on the progress of drug repositioning as a potential strategy for the treatment of mucosal parasites.