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.

Effect of the administration of Lactobacillus spp. strains on neonatal diarrhoea, immune parameters and pathogen abundance in pre-weaned calves.

Neonatal calf diarrhoea is one of the challenges faced by intensive farming, and probiotics are considered a promising approach to improve calves' health. The objective of this study was to evaluate the effect of potential probiotic lactobacilli on new-born dairy calves' growth, diarrhoea incidence, faecal score, cytokine expression in blood cells, immunoglobulin A (IgA) levels in plasma and faeces, and pathogen abundance in faeces. Two in vivo assays were conducted at the same farm in two annual calving seasons. Treated calves received one daily dose of the selected lactobacilli (Lactobacillus reuteri TP1.3B or Lactobacillus johnsonii TP1.6) for 10 consecutive days. A faecal score was recorded daily, average daily gain (ADG) was calculated, and blood and faeces samples were collected. Pathogen abundance was analysed by absolute qPCR in faeces using primers directed at Salmonella enterica, rotavirus, coronavirus, Cryptosporidium parvum and three Escherichia coli virulence genes (eae, clpG and Stx1). The faecal score was positively affected by the administration of both lactobacilli strains, and diarrhoea incidence was significantly lower in treated calves. No differences were found regarding ADG, cytokine expression, IgA levels and pathogen abundance. Our findings showed that oral administration of these strains could improve gastrointestinal health, but results could vary depending on the calving season, which may be related to pathogen seasonality and other environmental effects.

A new passive immune strategy based on IgY antibodies as a key element to control neonatal calf diarrhea in dairy farms.

BACKGROUND: Neonatal diarrhea remains one of the main causes of morbi-mortality in dairy calves under artificial rearing. It is often caused by infectious agents of viral, bacterial, or parasitic origin. Cows vaccination and colostrum intake by calves during the first 6 h of life are critical strategies to prevent severe diarrhea but these are still insufficient. Here we report the field evaluation of a product based on IgY antibodies against group A rotavirus (RVA), coronavirus (CoV), enterotoxigenic Escherichia coli, and Salmonella sp. This product, named IgY DNT, has been designed as a complementary passive immunization strategy to prevent neonatal calf diarrhea. The quality of the product depends on the titers of specific IgY antibodies to each antigen evaluated by ELISA. In the case of the viral antigens, ELISA antibody (Ab) titers are correlated with protection against infection in calves experimentally challenged with RVA and CoV (Bok M, et al., Passive immunity to control bovine coronavirus diarrhea in a dairy herd in Argentina, 2017), (Vega C, et al., Vet Immunol Immunopathol, 142:156-69, 2011), (Vega C, et al., Res Vet Sci, 103:1-10, 2015). To evaluate the efficiency in dairy farms, thirty newborn Holstein calves were randomly assigned to IgY DNT or control groups and treatment initiated after colostrum intake and gut closure. Calves in the IgY DNT group received 20 g of the oral passive treatment in 2 L of milk twice a day during the first 2 weeks of life. Animals were followed until 3 weeks of age and diarrhea due to natural exposure to infectious agents was recorded during all the experimental time. RESULTS: Results demonstrate that the oral administration of IgY DNT during the first 2 weeks of life to newborn calves caused a delay in diarrhea onset and significantly reduced its severity and duration compared with untreated calves. Animals treated with IgY DNT showed a trend towards a delay in RVA infection with significantly shorter duration and virus shedding compared to control calves. CONCLUSIONS: This indicates that IgY DNT is an effective product to complement current preventive strategies against neonatal calf diarrhea in dairy farms. Furthermore, to our knowledge, this is the only biological product available for the prevention of virus-associated neonatal calf diarrhea.

The Cryptosporidium parvum gp60 glycoprotein expressed in the ciliate Tetrahymena thermophila is immunoreactive with sera of calves infected with Cryptosporidium oocysts.

Cryptosporidium parvum is a protozoan parasite of the phylum Apicomplexa responsible for cryptosporidiosis in calves, a disease that causes significant diarrhea and impairs gain of body weight, generating important production losses. As to now, no effective drugs or vaccines are available for the treatment or prevention of bovine cryptosporidiosis. Several reports suggest that development of a vaccine to prevent cryptosporidiosis is feasible, but relatively few vaccine candidates have been characterized and tested. The most prominent C. parvum antigen is gp60, an O-glycosylated mucin-like protein tethered to the parasite membrane by a glycosylphosphatidylinositol (GPI) anchor. Gp60 has been shown to be involved in essential mechanisms for the survival of C. parvum, such as recognition, adhesion to, and invasion of host cells. This work was aimed at expressing gp60 in Tetrahymena thermophila, a ciliated protozoon with numerous advantages for the heterologous expression of eukaryotic proteins, as a first approach for the development of a recombinant vaccine for bovine cryptosporidiosis. T. thermophila-expressed gp60 localized to the protozoon cell surface and oral apparatus, and partitioned into the Triton X-114 detergent phase. This indicates that the protein entered the reticuloendothelial system of the ciliate, and suggests it contains a GPI-anchor. Homogenates of gp60-expressing T. thermophila cells were recognized by sera from calves naturally infected with C. parvum demonstrating their immunoreactivity. In summary, the heterologous expression of gp60, a C. parvum-encoded GPI-anchored protein, has been successfully demonstrated in the ciliate T. thermophila.

A highly sensitive method for detecting Cryptosporidium parvum oocysts recovered from source and finished water using RT-PCR directed to Cryspovirus RNA.

Sensitive detection of Cryptosporidium oocysts is important because the protozoan can cause clinical infection in humans at extremely low numbers. In the present study, 1.5 × 102, 1.0 × 103, or 1.0 × 104C. parvum oocysts were spiked into 10 l of source or finished water in triplicate followed by recovery using Envirochek HV sampling capsules. One subsample of the recovered oocysts was analyzed by commercial immunofluorescence assay (IFA), while a second subsample was subjected to DNA-RNA extraction, followed by RT-PCR using primers directed to the gene encoding Cryspovirus capsid. IFA analysis of Envirochek filter eluates of finished water detected oocysts at all 3 C. parvum oocyst doses, but only at the 1.0 × 103 and 1.0 × 104 doses in source water. Cryspovirus RT-PCR appeared to offer greater sensitivity than IFA because C. parvum oocysts were detected using this molecular technique in both source and finished water concentrates at all 3 spiking levels. A linear relationship was observed between log oocysts spiking dose and the relative intensity of the Cryspovirus RT-PCR signal for finished water, but not for source water. These data indicate that Cryspovirus RT-PCR is a sensitive method for detecting C. parvum oocysts in source and finished water.

Identification of novel vaccine candidates against cryptosporidiosis of neonatal bovines by reverse vaccinology.

The apicomplexan protozoan Cryptosporidium parvum is an important causative agent of diarrhea of neonatal bovines. Vaccination has been proposed as an advantageous strategy against cryptosporidiosis of calves since besides protection against disease it has also the potential to prevent dissemination of infective oocysts into the environment. Antigens anchored to the parasite surface via glycosylphosphatidylinositol (GPI) are implicated in host cell attachment and invasion and represent promising vaccine candidates. A reverse vaccinology approach was employed to (i) identify the GPI-anchored proteome of C. parvum using available web-based bioinformatic tools and (ii) characterize previously unrecognized novel vaccine antigens. Altogether, 14 putative GPI-anchored proteins could be determined of which CpH1 and CpSUB2 as well as GP60 were further characterized. Sequencing and comparison of GP60, CpH1, and CpSUB1 alleles amplified from different geographic isolates showed a high degree of conservation. All three antigens were recombinant expressed and immunoblotted using sera of 12 Cryptosporidium-infected calves sampled at age periods 1-11 and 12-28 days after birth. Specific antibody reactions against the studied antigens were detected in all analyzed calves, demonstrating their immunreactivity and expression, and recognition in vivo at an early stage of host infection. Besides the acknowledged GP60 vaccinogen, the presented reverse vaccinology approach reveals the additional vaccine candidates CpH1 and CpSUB1 for inclusion into a subunit vaccine formulation.

[Cryptosporidiosis and "One Health"]. / Criptosporidiosis y «Una Salud¼.

Cryptosporidiosis is a diarrheal disease (zoonotic) caused by the parasite Cryptosporidium, which infects the intestine. It occurs in humans and in different pets, farm and wild animals. Its role is underestimated and little is known about its epidemiology, due to the low interest it draws and the limited availability of diagnostic methods. The population health, both human and animal, is related to a close interaction of three factors: the environment, pathogens, and population, which determines the state of health and disease; some of the diseases considered as emerging, that appeared in the last years. The approach, known as "One Health", is a growing global strategy being adopted by a diversity of organizations and policy makers in response to the need for integrated approaches. This approach may be relevant to a wide range of global development goals. There is more and more talk about holistic health. It seems that we are more aware that health is not reduced to something purely biological, but affects the whole person. It is not only seeing the patient globally, it is starting from the complexity of the human being and the whole world crossed by vulnerability and interacting with the totality of the subjects.

Criptosporidiosis es una enfermedad diarreica (zoonótica) causada por el parásito Cryptosporidium, el cual infecta al intestino. Se producen tanto en el ser humano como en diferentes animales de granja, mascotas y animales silvestres. Se subestima su papel y poco se conoce sobre su epidemiología, debido al escaso interés que suscita y la limitada disponibilidad de métodos de diagnóstico. La salud de las poblaciones tanto humana como animal, están relacionadas con una estrecha interacción de tres factores: el ambiente, los agentes patógenos, y las poblaciones, lo cual determina el estado de salud y enfermedad. El enfoque, conocido como «Una Salud¼, es una estrategia global en crecimiento que se está adoptando por una diversidad de organizaciones y responsables de las políticas en respuesta a la necesidad de enfoques integrados. Este enfoque puede ser relevante para una amplia gama de objetivos mundiales de desarrollo. Cada vez se habla más de salud integral, de salud holística. Parece que somos más conscientes de que la salud no se reduce a algo puramente biológico, sino que afecta a toda la persona. No es sólo ver al enfermo globalmente, es partir de la complejidad del ser humano y del mundo entero atravesado por la vulnerabilidad e interaccionar con la totalidad de los sujetos.

Criptosporidiosis y una salud / Cryptosporidiosis and one health

RESUMEN Criptosporidiosis es una enfermedad diarreica (zoonótica) causada por el parásito Cryptosporidium, el cual infecta al intestino. Se producen tanto en el ser humano como en diferentes animales de granja, mascotas y animales silvestres. Se subestima su papel y poco se conoce sobre su epidemiología, debido al escaso interés que suscita y la limitada disponibilidad de métodos de diagnóstico. La salud de las poblaciones tanto humana como animal, están relacionadas con una estrecha interacción de tres factores: el ambiente, los agentes patógenos, y las poblaciones, lo cual determina el estado de salud y enfermedad. El enfoque, conocido como «Una Salud¼, es una estrategia global en crecimiento que se está adoptando por una diversidad de organizaciones y responsables de las políticas en respuesta a la necesidad de enfoques integrados. Este enfoque puede ser relevante para una amplia gama de objetivos mundiales de desarrollo. Cada vez se habla más de salud integral, de salud holística. Parece que somos más conscientes de que la salud no se reduce a algo puramente biológico, sino que afecta a toda la persona. No es sólo ver al enfermo globalmente, es partir de la complejidad del ser humano y del mundo entero atravesado por la vulnerabilidad e interaccionar con la totalidad de los sujetos.(AU)

ABSTRACT Cryptosporidiosis is a diarrheal disease (zoonotic) caused by the parasite Cryptosporidium, which infects the intestine. It occurs in humans and in different pets, farm and wild animals. Its role is underestimated and little is known about its epidemiology, due to the low interest it draws and the limited availability of diagnostic methods. The population health, both human and animal, is related to a close interaction of three factors: the environment, pathogens, and population, which determines the state of health and disease; some of the diseases considered as emerging, that appeared in the last years. The approach, known as "One Health", is a growing global strategy being adopted by a diversity of organizations and policy makers in response to the need for integrated approaches. This approach may be relevant to a wide range of global development goals. There is more and more talk about holistic health. It seems that we are more aware that health is not reduced to something purely biological, but affects the whole person. It is not only seeing the patient globally, it is starting from the complexity of the human being and the whole world crossed by vulnerability and interacting with the totality of the subjects.(AU)

Cryptosporidium spp. em idosos residentes em instituições de longa permanência no sul do Brasil / Cryptosporidium spp. in older adults living in long-term care facilities in southern Brazil

Introdução: A criptosporidiose é uma doença mundial, levando a quadros desde assintomáticos à diarreia grave. O presente estudo teve como objetivo detectar oocistos de Cryptosporidium spp. em idosos residentes em instituições de longa permanência na cidade de Lages, estado de Santa Catarina, Brasil, buscando avaliar a situação da parasitose, relativamente desconhecida na população idosa, foi avaliada por meio de um método diagnóstico fácil e de baixo custo, porém nem sempre realizado na rotina laboratorial. Métodos: Foram coletadas amostras dos exames Parasitológicos de fezes de 93 idosos residentes em instituições de longa permanência no período de setembro a novembro de 2015. Resultados: Os oocistos do protozoário foram detectados pela técnica de Ziehl-Neelsen modificada. A positividade para oocistos de Cryptosporidium spp. foi de 6,45% (6/93). Em relação ao sexo, a positividade foi de 8,9% (4/45) para mulheres e 2,1% (2/48) dos homens. Todos os pacientes tinham idade entre 60 e 70 anos. Em relação à consistência fecal, todas as amostras foram descritas como diarreia. Conclusão: A criptosporidiose é uma infecção parasitária importante e debilitante que afeta os idosos e precisa ser tratada. Os cuidadores devem estar cientes disso, para que tenham cuidados adequados, especialmente de higiene individual e uso de utensílios que podem transmitir a infecção aos demais idosos que residem nessas instituições. (AU)

Introduction: Cryptosporidiosis is a worldwide disease whose characteristics range from absence of symptoms to severe diarrhea. The present study aimed to detect Cryptosporidium spp. oocysts in older adults living in long-term care facilities in the city of Lages, state of Santa Catarina, Brazil. The situation of this parasitosis, which is relatively unknown in the older population, was evaluated using an easy and low-cost diagnostic method, although not always performed in laboratory routine. Methods: Stool samples were obtained from 93 older adults living in long-term care facilities from September to November 2015. Results: Protozoan oocysts were detected by the modified Ziehl-Neelsen technique. Positivity for Cryptosporidium spp. oocysts was 6.45% (6/93). Regarding sex, 8.9% (4/45) of women and 2.1% (2/48) of men were positive. All patients were aged between 60 and 70 years. Regarding fecal consistency, all samples were described as diarrhea. Conclusion: Cryptosporidiosis is an important, debilitating parasitic infection that affects older age and needs to be addressed. Caregivers must be aware of it in order to provide appropriate care, especially in terms of individual hygiene and use of tools that may transmit infection to other older adults living in these facilities. (AU)

Evaluation of recombinant Cryptosporidium hominis GP60 protein and anti-GP60 chicken polyclonal IgY for research and diagnostic purposes / Avaliação da proteína recombinante GP60 de Cryptosporidium hominis e da IgY policlonal anti-GP60 recombinante para uso em pesquisa e diagnóstico

In this study, a method for expressing Cryptosporidium hominis GP60 glycoprotein in Escherichia coli for production of polyclonal anti-GP60 IgY in chickens was developed aiming future studies concerning the diagnosis, prevention and treatment of cryptosporidiosis. The full-length nucleotide sequence of the C. hominis gp60 gene was codon-optimized for expression in E. coli and was synthesized in pET28-a vector. Subcloning was performed on several different strains of BL21 E. coli. Temperature, time and inducer IPTG concentration assays were also performed and analyzed using SDS-PAGE. The optimal conditions were observed at a temperature of 37 °C, with overnight incubation and 1 mM of IPTG. Purification was performed by means of affinity chromatography using the AKTA Pure chromatography system and the Hi-Trap HP column (GE Healthcare). The recombinant protein GP60 (rGP60) thus generated was used to immunize laying hens owing the production of polyclonal IgY. Western blot and indirect immunofluorescence showed that the polyclonal antibody was capable of binding to rGP60 and to Cryptosporidium parvum sporozoites, respectively. The rGP60 and the IgY anti-rGP60 generated in this study may be used as templates for research and for the development of diagnostic methods for cryptosporidiosis.(AU)

Neste trabalho, foi desenvolvido um método de expressão da glicoproteína GP60 de Cryptosporidium hominis em Escherichia coli visando produzir anticorpos IgY anti-GP60 em galinhas para utilização em estudos futuros com os objetivos de diagnóstico, prevenção e tratamento da criptosporidiose. A sequência completa de nucleotídeos do gene gp60 de C. hominis foi códon-otimizada para expressão em E. coli e sintetizada no vetor pET28-a. A subclonagem foi realizada em várias estirpes diferentes de E. coli BL21. Os ensaios de concentração do indutor IPTG, temperatura e tempo foram realizados e analisados por SDS-PAGE. As condições ótimas de expressão foram observadas em temperatura de 37 °C, incubação durante a noite e 1 mM de IPTG. A purificação da proteína foi realizada por cromatografia de afinidade utilizando o sistema de cromatografia AKTA Pure e a coluna Hi-Trap HP (GE Healthcare). A proteína recombinante GP60 (rGP60) foi utilizada para imunizar galinhas poedeiras para produzir IgY policlonal anti-rGP60. Verificou-se por Western blot e por imunofluorescência indireta que o anticorpo policlonal apresentou reatividade com a rGP60 e com esporozoítos de Cryptosporidium parvum, respectivamente. A rGP60 e a IgY anti-rGP60 geradas neste estudo podem ser utilizadas como modelos para o desenvolvimento de ensaios para pesquisa e diagnóstico da criptosporidiose.(AU)

Measurement of Elastic Modulus and Vickers Hardness of Surround Bone Implant Using Dynamic Microindentation - Parameters Definition

The clinical performance of dental implants is strongly defined by biomechanical principles. The aim of this study was to quantify the Vicker's hardness (VHN) and elastic modulus (E) surround bone to dental implant in different regions, and to discuss the parameters of dynamic microindantion test. Ten cylindrical implants with morse taper interface (Titamax CM, Neodent; 3.5 mm diameter and 7 mm a height) were inserted in rabbit tibia. The mechanical properties were analyzed using microhardness dynamic indenter with 200 mN load and 15 s penetration time. Seven continuous indentations were made distancing 0.08 mm between each other perpendicularly to the implant-bone interface towards the external surface, at the limit of low (Lp) and high implant profile (Hp). Data were analyzed by Student's t-test (a=0.05) to compare the E and VHN values obtained on both regions. Mean and standard deviation of E (GPa) were: Lp. 16.6 ± 1.7, Hp. 17.0 ± 2.5 and VHN (N/mm2): Lp. 12.6 ± 40.8, Hp. 120.1 ± 43.7. No statistical difference was found between bone mechanical properties of high and low profile of the surround bone to implant, demonstrating that the bone characterization homogeneously is pertinent. Dynamic microindantion method proved to be highly useful in the characterization of the individual peri-implant bone tissue.

O desempenho clínico de implantes dentais é fortemente definido por princípios biomecânicos. Este trabalho objetivou quantificar a Dureza Vickers (VHN) e módulo de elasticidade (E) do osso periimplantar e discutir parâmetros metodológicos de ensaio dinâmico de indentação. Foram utilizados 10 implantes de corpo cilíndrico com interface cone morse, (Titamax CM; Neodent, Curitiba, PR, Brasil), diâmetro de 3.5 mm e altura de 7 mm inseridos em tíbia de coelho recém obtidas após abate dos animais. As propriedades mecânicas foram analisadas usando penetrador dinâmico de microdureza Vickers (CSM Micro-Hardness Tester; CSM Instruments, Peseux, Switzerland) com carga de 200 mN e tempo de penetração de 15s. Foram feitas 7 indentações no osso cortical na base da rosca (Br) e na ponta da rosca (Pr) na direção perpendicular ao implante, com distância entre elas de 0,08 mm perpendicular a interface osso implante em direção a superfície esterna. Os dados foram analisados por meio de teste t-Student (P<0,05). O valores médios e desvio padrão de E (GPa) foram: Br. 16,6 ± 1,7A; Pr. 17,0 ± 2,5A e VHN (N/mm2): Br. 125,6 ± 40,8A; Pr. 120,1 ± 43,7A. Não houve diferença significativa entre as propriedades mecânicas avaliadas no osso na base e na ponta da rosca do implante, demonstrando que a caracterização desta estrutura de forma homogênea em análises computacionais é pertinente. O método de indentação dinâmica mostrou ser altamente útil na caracterização individualizada do tecido ósseo periimplantar.

Perception of tutors of dogs and cats of Recife city, Pernambuco State, Brazil, as to the risk factors infection Cryptosporidium spp / Percepção de tutores de cães e gatos da cidade do recife, estado de Pernambuco, Brasil, quanto aos fatores de risco à infecção por Cryptosporidium spp

Parasitic zoonoses are a public health problem and given the proximity between animals and humans, aimed to evaluate the perception of tutors of dogs and cats living to the city of Recife, state of Pernambuco, Brazil, as to infection by Cryptosporidium spp. and their risk factors. To this end, a questionnaire was applied, consisting of closed questions, the 71 tutors. The results were tabulated in Microsoft Excel® 2007 spreadsheet version to obtain the absolute and relative frequencies. Among the participants tutors, 16.9% (n = 12) were male and 83.1% (n = 59) were female. Their ages ranged from 18 to 75 years. Regarding cryptosporidiosis, only 11.3 % (n = 8) of the tutors reported having knowledge about the subject and 88.7% (n = 63) cited total nescience. A percentage of 100% (n = 71) was also achieved when questioned about the use of anticoccidial medications, responding not use them. On the possibility of the animals transmitting diseases to humans, they claimed to have that knowledge. Among the participants, 73.2% (n = 52) stated collect the feces of their pets. Tutors 80.2% (n = 57) reported that their animals had monitoring by a veterinarian. The concept of zoonosis was correctly answered by only 21.1% (n = 15) of them. The results showed a low level of knowledge about risk factors related to infection by Cryptosporidium spp., which exposes both the tutors and their animals as the collective health of the studied region to cryptosporidiosis.

As zoonoses parasitárias são um problema de saúde pública e diante da proximidade entre animais e humanos, objetivou-se avaliar a percepção de tutores de cães e gatos residentes à cidade do Recife, estado de Pernambuco, Brasil, quanto à infecção por Cryptosporidium spp. e seus fatores de risco. Para tanto, foi aplicado um questionário, composto por perguntas fechadas, a 71 tutores. Os resultados foram tabulados em planilhas do Microsoft Excel® versão 2007 para obtenção das frequências absolutas e relativas. Entre os tutores participantes, 16,9% (n = 12) eram do sexo masculino e 83,1% (n = 59) do sexo feminino. Suas respectivas idades variaram de 18 a 75 anos. Em relação à criptosporidiose, apenas 11,3% (n = 8) dos tutores responderam ter conhecimento sobre o assunto e 88,7% (n = 63) citaram uma total insciência. Um percentual de 100% (n = 71) foi igualmente alcançado ao serem questionados sobre o uso de medicações anticoccidianas, respondendo não utilizá-las. Sobre a possibilidade de os animais transmitirem doenças para o ser humano, afirmaram ter esse conhecimento. Dentre os participantes, 73,2% (n = 52) afirmaram recolher as fezes de seus animais. Dos tutores 80,2% (n = 57) relataram que seus animais possuíam acompanhamento por médico veterinário. O conceito de zoonose foi corretamente respondido por apenas 21,1% (n = 15) deles. Os resultados obtidos evidenciaram um reduzido nível de conhecimento sobre fatores de risco relacionados à infecção por Cryptosporidium spp., o que expõe tanto os tutores e seus animais quanto a saúde coletiva da região estudada à criptosporidiose.

Perception of tutors of dogs and cats of Recife city, Pernambuco State, Brazil, as to the risk factors infection Cryptosporidium spp / Percepção de tutores de cães e gatos da cidade do recife, estado de Pernambuco, Brasil, quanto aos fatores de risco à infecção por Cryptosporidium spp

Parasitic zoonoses are a public health problem and given the proximity between animals and humans, aimed to evaluate the perception of tutors of dogs and cats living to the city of Recife, state of Pernambuco, Brazil, as to infection by Cryptosporidium spp. and their risk factors. To this end, a questionnaire was applied, consisting of closed questions, the 71 tutors. The results were tabulated in Microsoft Excel® 2007 spreadsheet version to obtain the absolute and relative frequencies. Among the participants tutors, 16.9% (n = 12) were male and 83.1% (n = 59) were female. Their ages ranged from 18 to 75 years. Regarding cryptosporidiosis, only 11.3 % (n = 8) of the tutors reported having knowledge about the subject and 88.7% (n = 63) cited total nescience. A percentage of 100% (n = 71) was also achieved when questioned about the use of anticoccidial medications, responding not use them. On the possibility of the animals transmitting diseases to humans, they claimed to have that knowledge. Among the participants, 73.2% (n = 52) stated collect the feces of their pets. Tutors 80.2% (n = 57) reported that their animals had monitoring by a veterinarian. The concept of zoonosis was correctly answered by only 21.1% (n = 15) of them. The results showed a low level of knowledge about risk factors related to infection by Cryptosporidium spp., which exposes both the tutors and their animals as the collective health of the studied region to cryptosporidiosis.(AU)

As zoonoses parasitárias são um problema de saúde pública e diante da proximidade entre animais e humanos, objetivou-se avaliar a percepção de tutores de cães e gatos residentes à cidade do Recife, estado de Pernambuco, Brasil, quanto à infecção por Cryptosporidium spp. e seus fatores de risco. Para tanto, foi aplicado um questionário, composto por perguntas fechadas, a 71 tutores. Os resultados foram tabulados em planilhas do Microsoft Excel® versão 2007 para obtenção das frequências absolutas e relativas. Entre os tutores participantes, 16,9% (n = 12) eram do sexo masculino e 83,1% (n = 59) do sexo feminino. Suas respectivas idades variaram de 18 a 75 anos. Em relação à criptosporidiose, apenas 11,3% (n = 8) dos tutores responderam ter conhecimento sobre o assunto e 88,7% (n = 63) citaram uma total insciência. Um percentual de 100% (n = 71) foi igualmente alcançado ao serem questionados sobre o uso de medicações anticoccidianas, respondendo não utilizá-las. Sobre a possibilidade de os animais transmitirem doenças para o ser humano, afirmaram ter esse conhecimento. Dentre os participantes, 73,2% (n = 52) afirmaram recolher as fezes de seus animais. Dos tutores 80,2% (n = 57) relataram que seus animais possuíam acompanhamento por médico veterinário. O conceito de zoonose foi corretamente respondido por apenas 21,1% (n = 15) deles. Os resultados obtidos evidenciaram um reduzido nível de conhecimento sobre fatores de risco relacionados à infecção por Cryptosporidium spp., o que expõe tanto os tutores e seus animais quanto a saúde coletiva da região estudada à criptosporidiose.(AU)

Assessing the infection risk of Giardia and Cryptosporidium in public drinking water delivered by surface water systems in Sao Paulo State, Brazil.

A survey of Giardia and Cryptosporidium was conducted in surface water used as drinking water sources by public water systems in four densely urbanized regions of Sao Paulo State, Brazil. A Quantitative Microbial Risk Assessment, based on protozoa concentrations, was performed to estimate the probability of protozoa infection associated with drinking water ingestion. A total of 206 source water samples were analyzed over a 24 month period using the USEPA Method 1623. The risk of infection was estimated using an exponential dose response model, children and adults exposure and a gamma distribution for (oo)cyst concentrations with three scenarios for treating censored data. Giardia was detected in 102 of the samples, and 19 of them were also positive for Cryptosporidium, with maximum concentrations of 97.0 cysts/L and 6.0 oocysts/L, respectively. Risk distributions were similar for the three scenarios. In the four regions, the estimated risk of Giardia infection per year, for adults and children, ranged from 0.29% to 2.47% and from 0.08% to 0.70%, respectively. Cryptosporidium risk infection varied from 0.15% to 0.29% for adults and from 0.04% to 0.08% for children. In both cases, the calculated risk surpassed the risk of infection of 10(-4) (1:10,000) defined as tolerable by USEPA for a yearly exposure. The probability of Giardia infection was very close to the rates of acute diarrheic disease for adults (1% to 3%) but lower for children (2% to 7%). The daily consumption of drinking water was an important contributing factor for these differences. The Microbiological Risk Assessment carried out in this study provides an indication of infection risks by Giardia and Cryptosporidium in the population served by these source waters. Strategies for source water protection and performance targets for the water treatment should be established to achieve the required level of public health risk.

[Cryptosporidiosis: an emerging zoonosis]. / Criptosporidiosis: una zoonosis emergente.

The genus Cryptosporidium, responsible for producing cryptosporidiosis, includes several species. Humans and livestock are the main sources of infection. Waterborne cryptosporidiosis outbreaks are associated with drinking water. The infective parasite stage is the oocyst, which is resistant to conventional potabilization treatments. In immunocompetent hosts it produces acute, self-limiting diarrhoea. In immunocompromised people, it could develop severe, life-threatening pattern forms of the infection. People with AIDS are especially susceptible to these clinical forms. Cryptosporidium infections are also considered a major cause of morbimortality in calves, which leads to important economic losses. In the last years, there has been an increase of patients suffering from different causes of immunosuppression, and the need to find an effective therapy against Cryptosporidium has become greater. In spite of the many attempts of the pharmaceutical industry to develop an effective antiparasitic agent to treat cryptosporidiosis, this infection and its clinical consequences still constitute a major public health problem. This article analizes the taxonomy, morphology, biology and life cycle of Cryptosporidium. Clinical, immunological, epidemiological features and diagnosis of cryptosporidiosis are also included. Treatment and prevention of the infection are discussed, and future tendencies are suggested for this emerging parasitic infection.

Sheep management and cryptosporidiosis in central Mexico.

To establish the association between sheep farm management practices and cryptosporidiosis in Central Mexico, 37 farms were invited and questionnaires answered by owners. Faeces samples were taken from lambs under 3 months old and ewes in the farms. Parasites were identified by modified Ziehl-Neelsen stain. Questionnaire included: general farm information, facilities characteristics, facilities cleaning measures, water use and farm and animal management practices. Farms were grouped in a low ( Pound 3,37%), medium (3.38-49.9%) and high basis ((3)50%) according to prevalence. Associations (OR) were calculated according to positivity, compared by chi-square test. A general prevalence of 34,33% was found. Five variables showed statistical significance (p<0.05). Associations found were: use of bed, grazing place, and bottle cleaning. The only association found between management practices and cryptosporidiosis that could represent a risk was the grazing place. This report is the first one in the country regarding predisposing conditions for cryptosporidiosis.

[The action of different coagulants to remove Cryptosporidium during the process of water treatment]. / Acción de distintos coagulantes para la eliminación de Cryptosporidium spp. en el proceso de potabilización del agua.

Cryptosporidium is one of the microorganisms of main concern from the point of view of Public Health, being a priority problem for water treatment plants and water regulatory institutions. Due to its small size and resistance to chlorination, Cryptosporidium removal during the process of drinking water treatment is a hard task. The effectiveness of different coagulants commonly used in the process of removal of oocysts was analyzed. The technique used was the Jar Test. It was found that: 1) coagulants with the addition of polimeric coadjuvants produce over 2 logs of oocyst removal; 2) a low value in turbidity does not necessarily mean optimal parasite removal, and 3) the addition of polyelectrolites to ferric chloride diminishes variability, both in final turbidity and Cryptosporidium removal.

Acción de distintos coagulantes para la eliminación de Cryptosporidium spp. en el proceso de potabilización del agua / The action of different coagulants to remove Cryptosporidium during the process of water treatment

Cryptosporidium es uno de los microorganismos de mayor interés desde el punto de vista de la Salud Pública y constituye un problema prioritario para las plantas y organismos reguladores de agua. Debido a su pequeño tamaño y a su resistencia a la cloración, la eliminación por el proceso de potabilización es una tarea compleja. En este trabajo se analizó la efectividad de distintos coagulantes utilizados comúnmente en tal proceso para lograr la remoción de los ooquistes. Se trabajó con la prueba de jarras (Jar Test). Se halló que: 1) Los coagulantes con agregado de polímeros coadyuvantes producen remociones de ooquistes superiores a 2 log. 2) Un valor bajo de turbiedad no asegura una remoción óptima de los parásitos. 3) La adición de polielectrolitos al cloruro férrico disminuye la variabilidad tanto en la turbiedad final como en la eliminación de Cryptosporidium.

Cryptosporidium is one of the microorganisms of main concern from the point of view of Public Health, being a priority problem for water treatment plants and water regulatory institutions. Due to its small size and resistance to chlorination, Cryptosporidium removal during the process of drinking water treatmentis a hard task. The effectiveness of different coagulants commonly used in the process of removal of oocysts was analyzed. Thetechnique used was the Jar Test. It was found that: 1) coagulants with the addition of polimeric coadjuvants produce over 2 logs of oocyst removal; 2) a low value in turbidity does not necessarily mean optimal parasite removal, and 3) the addition of polyelectrolites to ferric chloride diminishes variability, both in final turbidity and Cryptosporidium removal.

III Taller Internacional de Infecciones por Clamidias, Brucelas y Micobacterias en Humanos y Animales: Programa y resúmenes de trabajo

Resúmenes de los trabajos presentados en el taller, y organizados en los siguientes módulos: Brucelosis, Micobacteriosis, Clamidiosis, y Criptosporidiosis; que tratan aspectos básicos y epidemiológicos de cada infección