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.

Cryptosporidium parvum in captive primates of Parque Municipal Danilo Galafassi, Paraná, Brazil / Cryptosporidium parvum em primatas cativos do Parque Municipal Danilo Galafassi, Paraná, Brasil

A wide variety of terrestrial and aquatic animal species have been identified as hosts of species and genotypes of Cryptosporidium spp., which are important pathogens, however, little is known about their distribution in wild populations. Recent studies associating parasitological findings and molecular techniques have provided a new insight into host specificity and its potential transmission to humans.The objective of this study was to investigate the presence of Cryptosporidium spp. in feces of Callithrixsp. and Ateles paniscus, identify the species, and evaluate their phylogenetic relationships with other representatives of the genus. Four samples of feces were collected from an enclosure where three Callithrix jacchus and one Callithrix penicillate live; in addition, five samples were collected from an enclosure of an Ateles paniscus from Parque Municipal Danilo Galafassi, located in the city of Cascavel-PR. These samples were sent to the UFPR Biotechnology Laboratory, where the modified Ziehl-Neelsen staining technique was performed on microscope slides with fecal smear. Positive samples were submitted to DNA purification, extraction, PCR, and sequencing of the nuclear SSU rRNA region. Phylogenetic analysis based on Maximum Parcimony and Bayesian Inference were performed. Fifty percent (2: 4) of the feces samples from the enclosure of the Callithrix spp. and 60 % (3: 5) of samples from the Ateles paniscus enclosure were positive for Cryptosporidium spp. The phylogenetic analysis showed that the parasite found in both species of primates was recovered nested with others genotypes of C. parvum, and the genotype found in Callithrix spp. has high similarity with that one founded in several domestic animals. This is the first report of C. parvum in A. paniscus. Because it is an important zoonosis which does not have treatment, preventive measures must be adopted to avoid the spread of the disease.(AU)

Uma grande variedade de espécies animais terrestres e aquáticas tem sido identificada como hospedeiros de espécies e genótipos de Cryptosporidium spp., que são importantes agentes patogênicos, mas pouco se conhece sobre a sua distribuição nas populações silvestres. Estudos recentes associando achados parasitológicos e técnicas moleculares têm proporcionado uma nova visão em relação à especificidade do hospedeiro e seu potencial de transmissão para o homem. O objetivo desse estudo foi pesquisar a presença de Cryptosporidium spp. em fezes de Callithrix spp. e Ateles paniscus, identificar a espécie e avaliar o seu relacionamento filogenético com outros representantes do gênero. Foram coletadas quatro amostras de fezes de um recinto onde convivem três Callithrix jacchus e um Callithrix penicillata e cinco amostras de um recinto onde vive um Ateles paniscus do Parque Municipal Danilo Galafassi localizado na cidade de Cascavel-PR. As amostras foram enviadas ao Laboratório de Biotecnologia da UFPR onde foi realizada a técnica de coloração de Ziehl-Neelsen modificado em lâminas de esfregaço de fezes. As amostras positivas foram submetidas à purificação, extração de DNA, PCR, e sequenciamento da região nuclear SSU rRNA. Foram realizadas análises filogenéticas baseadas em Máxima Parcimônia e Inferência Bayesiana. Cinquenta por cento (2:4) das amostras de fezes do recinto dos Callithrix...(AU)

Comparison of the effectiveness of halofuginone lactate and paromomycin in the treatment of calves naturally infected with Cryptosporidium parvum

Background: Cryptosporidiosis is one of the most important problems among neonatal ruminants. Cryptosporidium parvum is the agent causing the disease. Cryptosporidium infection, responsible for diarrhea, dehydration, weight loss and death in neonatal ruminants, leads to significant economic losses for producers. In calves naturally or experimentally infected with cryptosporidiosis, many agents have been reported to have therapeutic and protective effects. The objective of this study was to compare the effectiveness of halofuginone lactate and paromomycin in the treatment of calves naturally infected with Cryptosporidium parvum.Materials, Methods & Results: Twenty calves between 7 and 20 days of age and naturally infected were included in the study. There were two different treatment groups in the study. The first group of calves were administered 100 µg/kg/day halofuginone lactate for 7 days and the second group of calves were administered 100 mg/kg/day paromomycin for 7 days. In addition, antibiotic and vitamin C were applied to all calves. Jugular venous blood samples were obtained pre-treatment and on the 7th day after the treatment. Routine clinical examinations of the calves were performed on days 0, 1, 3, 5 and 7 and rectal stool samples were collected for the detection of Cryptosporidium oocysts. Severe diarrhea, dehydration, depression and weight loss were observed in calves in both treatment groups. A significant decrease (P < 0.05) was observed for both groups in oocyst count on days 3, 5 and 7 compared to days 0 and 1. Improvements in blood parameters, stool characters and clinical scoring of both groups were observed in post-treatment. Discussion: Many pharmaceuticals or compounds have been tested for animal with cryptosporidiosis and only very few of them have shown a partial protective activity in ruminants when used prophylactically. Halofuginone lactate and paromomycin are commonly recommended as both therapeutic and protective agents.[...]

Comparison of the effectiveness of halofuginone lactate and paromomycin in the treatment of calves naturally infected with Cryptosporidium parvum

Background: Cryptosporidiosis is one of the most important problems among neonatal ruminants. Cryptosporidium parvum is the agent causing the disease. Cryptosporidium infection, responsible for diarrhea, dehydration, weight loss and death in neonatal ruminants, leads to significant economic losses for producers. In calves naturally or experimentally infected with cryptosporidiosis, many agents have been reported to have therapeutic and protective effects. The objective of this study was to compare the effectiveness of halofuginone lactate and paromomycin in the treatment of calves naturally infected with Cryptosporidium parvum.Materials, Methods & Results: Twenty calves between 7 and 20 days of age and naturally infected were included in the study. There were two different treatment groups in the study. The first group of calves were administered 100 µg/kg/day halofuginone lactate for 7 days and the second group of calves were administered 100 mg/kg/day paromomycin for 7 days. In addition, antibiotic and vitamin C were applied to all calves. Jugular venous blood samples were obtained pre-treatment and on the 7th day after the treatment. Routine clinical examinations of the calves were performed on days 0, 1, 3, 5 and 7 and rectal stool samples were collected for the detection of Cryptosporidium oocysts. Severe diarrhea, dehydration, depression and weight loss were observed in calves in both treatment groups. A significant decrease (P < 0.05) was observed for both groups in oocyst count on days 3, 5 and 7 compared to days 0 and 1. Improvements in blood parameters, stool characters and clinical scoring of both groups were observed in post-treatment. Discussion: Many pharmaceuticals or compounds have been tested for animal with cryptosporidiosis and only very few of them have shown a partial protective activity in ruminants when used prophylactically. Halofuginone lactate and paromomycin are commonly recommended as both therapeutic and protective agents.[...](AU)

Serum concentrations of acute phase proteins and immunoglobulins of calves with rotavirus diarrhea / Concentração sérica de proteínas de fase aguda e imunoglobulinas em bezerros acometidos por diarreia por rotavírus

The aim of the present study was to characterize changes in acute phase protein levels according to the occurrence of rotavirus diarrhea in calves in the first month of life. Blood and fecal samples were taken before colostrum intake and at 1, 2, 7, 15, 21 and 30 days of age from 24 Holstein calves allotted in three experimental groups: calves that did not present diarrhea (group A), calves that presented diarrhea, but tested negative for rotavirus in feces (group B), and calves that presented diarrhea and tested positive for rotavirus in feces (group C) (experiment 1). When the animals presented episodes of diarrhea, blood and fecal samples were taken at 24-hour intervals until the end of clinical signs (experiment 2). Serum proteins were separated by SDS-PAGE technique and rotavirus in feces was detected by PAGE. Data of experiment 1 were analyzed by ANOVA and Tukey's test, considered significant at P<0.05. Data of experiment 2 were subjected to the HSD test. Total protein, globulins, and IgG concentrations were lower in group C than in groups A and B. Ceruloplasmin and transferrin levels were higher in group C than in groups A and B. Serum concentrations of haptoglobin and α1-acid glycoprotein did not differ significantly between groups throughout the experimental period. Calves presented diarrhea between 10.4 and 14.6 days of age in group B, and between 10.3 and 14.6 days of age in group C. In the moments of diarrhea manifestation, least square means of IgA, haptoglobin and α1-acid glycoprotein concentrations did not differ significantly between groups B and C, but ceruloplasmin and transferrin concentrations were higher in group C than in group B, as opposed to what occurred with IgG levels. These findings show that optimizing passive immunity transfer of immunoglobulins decrease the likelihood of calves developing diarrhea caused by rotavirus. In addition, ceruloplasmin presents characteristics of a biomarker of rotavirus infection in calves.(AU)

O objetivo do presente estudo foi avaliar alterações nos teores de proteínas de fase aguda de acordo com a ocorrência de diarreia por rotavírus em bezerros no decorrer do primeiro mês de vida. Amostras de sangue e fezes de 24 bezerros da raça Holandesa foram coletadas antes da ingestão de colostro e com um, dois, sete, quinze, vinte um e trinta dias de idade, sendo os bezerros alocados em três grupos: bezerros que não apresentaram diarreia (grupo A), bezerros que apresentaram diarreia, mas foram negativos para a detecção de rotavírus nas fezes (grupo B) e bezerros que apresentaram diarreia e foram positivos para detecção de rotavírus nas fezes (grupo C) (experimento 1). Sempre que os animais apresentavam episódio de diarreia, amostras de sangue e fezes eram coletadas em intervalos de 24 horas até o término dos sinais clínicos (experimento 2). As proteínas séricas foram separadas por meio da técnica de SDS-PAGE e a pesquisa de rotavírus nas fezes foi realizada por meio da técnica de PAGE. Os resultados do experimento 1 foram analisados por meio de ANOVA e do teste de Tukey, considerado significativo quando P<0,05. Os dados do experimento 2 foram submetidos ao teste HSD. Os teores de proteína total, globulinas e IgG foram menores no grupo C que nos grupos A e B, os teores de ceruloplasmina e transferrina foram maiores no grupo C que nos grupos A e B e as concentrações séricas de haptoglobina e α1-glicoproteína ácida não diferiram significativamente entre grupos. Os bezerros manifestaram diarreia, em média, com 10,4 a 14,6 dias de idade no grupo B e com 10,3 a 14,6 dias de idade no grupo C. Nos momentos de manifestação de diarreia, os teores de IgA, haptoglobina e α1-glicoproteína ácida não diferiram significativamente entre os grupos B e C, mas os teores de ceruloplasmina e transferrina foram maiores no grupo C que no grupo B, oposto ao verificado para o teor de IgG. Esses achados mostram que a otimização da transferência de imunidade passiva de imunoglobulinas reduz a probabilidade de os animais apresentarem diarreia por rotavírus. Adicionalmente, a ceruloplasmina apresenta características de um biomarcador da infecção por rotavírus em bezerros.(AU)

Serum concentrations of acute phase proteins and immunoglobulins of calves with rotavirus diarrhea / Concentração sérica de proteínas de fase aguda e imunoglobulinas em bezerros acometidos por diarreia por rotavírus

The aim of the present study was to characterize changes in acute phase protein levels according to the occurrence of rotavirus diarrhea in calves in the first month of life. Blood and fecal samples were taken before colostrum intake and at 1, 2, 7, 15, 21 and 30 days of age from 24 Holstein calves allotted in three experimental groups: calves that did not present diarrhea (group A), calves that presented diarrhea, but tested negative for rotavirus in feces (group B), and calves that presented diarrhea and tested positive for rotavirus in feces (group C) (experiment 1). When the animals presented episodes of diarrhea, blood and fecal samples were taken at 24-hour intervals until the end of clinical signs (experiment 2). Serum proteins were separated by SDS-PAGE technique and rotavirus in feces was detected by PAGE. Data of experiment 1 were analyzed by ANOVA and Tukey's test, considered significant at P 0.05. Data of experiment 2 were subjected to the HSD test. Total protein, globulins, and IgG concentrations were lower in group C than in groups A and B. Ceruloplasmin and transferrin levels were higher in group C than in groups A and B. Serum concentrations of haptoglobin and 1-acid glycoprotein did not differ significantly between groups throughout the experimental period. Calves presented diarrhea between 10.4 and 14.6 days of age in group B, and between 10.3 and 14.6 days of age in group C. In the moments of diarrhea manifestation, least square means of IgA, haptoglobin and 1-acid glycoprotein concentrations did not differ significantly between groups B and C, but ceruloplasmin and transferrin concentrations were higher in group C than in group B, as opposed to what occurred with IgG levels. These findings show that optimizing passive immunity transfer of immunoglobulins decrease the likelihood of calves developing diarrhea caused by rotavirus. In addition, ceruloplasmin presents characteristics of a biomarker of rotavirus infection in calves.(AU)

O objetivo do presente estudo foi avaliar alterações nos teores de proteínas de fase aguda de acordo com a ocorrência de diarreia por rotavírus em bezerros no decorrer do primeiro mês de vida. Amostras de sangue e fezes de 24 bezerros da raça Holandesa foram coletadas antes da ingestão de colostro e com um, dois, sete, quinze, vinte um e trinta dias de idade, sendo os bezerros alocados em três grupos: bezerros que não apresentaram diarreia (grupo A), bezerros que apresentaram diarreia, mas foram negativos para a detecção de rotavírus nas fezes (grupo B) e bezerros que apresentaram diarreia e foram positivos para detecção de rotavírus nas fezes (grupo C) (experimento 1). Sempre que os animais apresentavam episódio de diarreia, amostras de sangue e fezes eram coletadas em intervalos de 24 horas até o término dos sinais clínicos (experimento 2). As proteínas séricas foram separadas por meio da técnica de SDS-PAGE e a pesquisa de rotavírus nas fezes foi realizada por meio da técnica de PAGE. Os resultados do experimento 1 foram analisados por meio de ANOVA e do teste de Tukey, considerado significativo quando P 0,05. Os dados do experimento 2 foram submetidos ao teste HSD. Os teores de proteína total, globulinas e IgG foram menores no grupo C que nos grupos A e B, os teores de ceruloplasmina e transferrina foram maiores no grupo C que nos grupos A e B e as concentrações séricas de haptoglobina e 1-glicoproteína ácida não diferiram significativamente entre grupos. Os bezerros manifestaram diarreia, em média, com 10,4 a 14,6 dias de idade no grupo B e com 10,3 a 14,6 dias de idade no grupo C. Nos momentos de manifestação de diarreia, os teores de IgA, haptoglobina e 1-glicoproteína ácida não diferiram significativamente entre os grupos B e C, mas os teores de ceruloplasmina e transferrina foram maiores no grupo C que no grupo B, oposto ao verificado para o teor de IgG. Esses achados mostram que a otimização da transferência de imunidade passiva de imunoglobulinas reduz a probabilidade de os animais apresentarem diarreia por rotavírus. Adicionalmente, a ceruloplasmina apresenta características de um biomarcador da infecção por rotavírus em bezerros.(AU)

Parasitological and molecular detection of cryptosporidium parvum in rheas (Rhea americana)

Background: The rhea (Rhea americana) is a Brazilian wild bird that produce meat, leather and feathers of excellent quality. Rhea production has been increasing every day in Brazil due to many favorable conditions for breeding and there are also large native populations in various regions of the country. The incidence of parasites is a limiting factor when raising many animals, and rheas are not an exception. The occurrence of Cryptosporidium spp in captive rheas in a Brazilian zoo and Spain was described. However, little is known about cryptosporidiosis in rhea, which makes the need for further studies. Thus, this study aimed to detect Cryptosporidium parvum in rheas from the South of Brazil. Case: This study evaluated two properties located in Southern Brazil. Property A was located in Rio Rufi no, Santa Catarina State, Brazil and it had 40 rheas for commercial purposes. Property B was located in Santa Maria, a small town in Rio Grande do Sul State, Brazil and it had 10 rheas. Fresh fecal samples were collected and kept refrigerated from adult birds (n = 4) and chicks (n = 2) from property A, and chicks (n = 3) of three months of age from property B. Samples were analyzed by the method of direct examination, followed by centrifugal flotation with zinc sulfate. Only the centrifugal flotation technique allowed the observation of Cryptosporidium spp. oocysts in three adults and one chick. Fecal samples were stored in ethanol and analyzed by PCR for C. parvum, all being positive for this protozoan. Feces contaminated by C. parvum oocysts from one young rhea was used to inoculate two mice orally (BALB/c), previously confirmed protozoan free by faecal examination and PCR. Feces from inoculated mice were collected on days 1, 3 e 5 post-inoculation for analysis by the centrifugal flotation technique. After five days of inoculation all mice presented diarrhea and high numbers of oocysts of protozoan in their feces. Discussion: Cryptosporidiosis can evolve into severe diarrhea, followed by abdominal cramps, anorexia, vomiting, dehydration, nausea and fever in different animals. However the incubation period of this disease in rheas is unknown because this is only the third report of cryptosporidiosis in this wild bird. The Cryptosporidium spp. is an obligate parasite of vertebrates, and its colonization occurs at the periphery of the intestinal cells of the host and it may cause atrophy of these structures and enteritis, but these findings have not been described in rheas yet. The species of Cryptosporidium that are mainly reported in birds are Cryptosporidium meleagridis, Cryptosporidium baileyi and Cryptosporidium galli. In our study, the molecular analysis was performed in order to identify the protozoan, being detected the C. parvum, a zoonotic agent reported in several mammals. The rhea is wild birds with habits similar to ostriches. Already, in studies have reported the occurrence of Cryptosporidium in ostriches; however the species identified were different from that described in this study. Based on these results, we conclude that rhea may be parasitized by C. parvum, an important zoonotic parasite. Prevalence studies should be conducted in this area to estimate the role and impact of rhea as reservoirs and disseminators of this zoonotic parasite.

Detection of Cryptosporidium parvum oocysts in calf fecal samples by direct immunofluorescence assay / Detecção de oocistos de Cryptosporidium parvum em amostras fecais de bezerros pela reação de imunofluorescência direta

The aim of this study was to produce a conjugate containing anti-Cryptosporidium parvum polyclonal antibodies and standardize a Direct Immunofluorescence Assay (DIF) for detecting C. parvum oocysts in fecal samples from calves. In order to obtain anti-C. parvum polyclonal antibodies, two New Zealand rabbits were immunized with a purified solution of C. parvum oocysts and Freund's adjuvant. Purification of the immunoglobulin G (IgG) fraction was performed by means of precipitation in ammonium sulfate and chromatography using a DEAE-cellulose column. The anti-C. parvum polyclonal antibody titer was determined by means of the enzyme-linked immunosorbent assay (ELISA). The rabbit anti-C. parvum IgG fraction was conjugated with fluorescein isothiocyanate and standardization of the DIF was performed using various dilutions of conjugate on slides positive for C. parvum oocysts. The cross-reactivity of the anti-C. parvum conjugate was tested using oocysts of Cryptosporidium serpentis, Cryptosporidium andersoni, Escherichia coli, Eimeria sp., and Candida sp. An anti-C. parvum conjugate was successfully produced, thus allowing standardization of DIF for detection of Cryptosporidium oocysts in fecal samples. Cross-reactivity of anti-C. parvum polyclonal antibodies with C. andersoni and C. serpentis was also observed.(AU)

O objetivo deste estudo foi produzir um conjugado contendo anticorpos policlonais anti-Cryptosporidium parvum e padronizar a Reação de Imunofluorescência Direta (RID), para detecção de oocistos de C. parvum em amostras fecais de bezerros. Para produção de anticorpos policlonais anti-C. parvum, dois coelhos da raça Nova Zelândia foram imunizados com uma solução purificada de oocistos de C. parvum e adjuvante de Freund. A purificação da fração de imunoglobulina G (IgG) foi realizada por meio de precipitação em sulfato de amônio e cromatografia em coluna de DEAE celulose. A titulação dos anticorpos policlonais anti-C. parvum foi determinada por meio de ensaio imunoenzimático (ELISA). A fração IgG de coelho anti-C. parvum foi conjugada com isotiocianato de fluoresceína, e a padronização da RID foi feita utilizando-se várias diluições do conjugado, em lâminas positivas para C. parvum. Foi pesquisada também a presença de reatividade cruzada do conjugado anti-C. parvum com C. serpentis, C. andersoni, Escherichia coli, Eimeria sp. e Candida sp.. A produção do conjugado anti-C. parvum foi bem sucedida, sendo possível a padronização da RID para detecção de oocistos em fezes. Foi também observada reatividade cruzada dos anticorpos policlonais anti-C. parvum, com C. andersoni e C. serpentis.(AU)

Risk factors associated with the occurrence of Cryptosporidium parvum infection in calves / Fatores de risco associados à ocorrência de infecção por Cryptosporidium parvum em bezerros

Cryptosporidium parvum oocysts were detected in feces of dairy calves raised in Rio de Janeiro State and the risk factors involved in the infection were determined. A hundred calves aging up to 12-month-old from 13 dairy farms were sampled. Polymerase chain reaction was used to detect the presence of oocysts. The zoonotic C. parvum species was detected in 45 percent animals. Statistical risk factors analyses revealed an association between infection and animals raised in technical systems such as the use of milking equipment, milking cooler, and water trough(P<0.05).(AU)

Detectaram-se oocistos de Cryptosporidium parvum em fezes de bezerros leiteiros no estado do Rio de Janeiro e analisaram-se os fatores de risco envolvidos na infecção dos animais. Cem bezerros com idades de 0 a 12 meses, provenientes de 13 propriedades rurais, foram amostrados, e suas fezes examinadas pela reação em cadeia da polimerase para a detecção dos oocistos. A espécie zoonótica C. parvum foi detectada em 45 por cento dos animais. As análises estatísticas dos fatores de risco revelaram haver associação entre infecção e animais criados em propriedades tecnificadas, que usam ordenha mecanizada, resfriamento de leite e fazendas que continham reservatórios de água à disposição dos animais (P<0,05).(AU)

Oocistos deCryptosporidium parvum em fezes de bezerro infectado experimentalmente / Cryptosporidium parvum oocysts in the feces of experimentally infected calf

A dose of 5.0x106 Cryptosporidium parvum oocysts was inoculated in a newborn calf. After the inoculation, the feces were daily collected and the presence of oocysts was examined on slides using 0.17% green malachite dye. The total yield reached 1.5x1010 oocysts, with a peak production on the 7th day, confirming the infectious process and the role of calf infection in the potential risk for environmental contamination. (AU)

Patterns of shedding of cryptosporidial oocysts by ewes and lambs kept indoors / Eliminação de oocistos de Cryptosporidium parvum por ovelhas e cordeiros confinados

Este experimento foi realizado com o objetivo de determinar a variação na eliminação de oocistos de Cryptosporidium parvum nas fezes de cordeiros e ovelhas, mantidos confinados do nascimento a desmama, em uma criação localizada em Botucatu -SP. Um grupo de 20 ovelhas da raça Ile de France em final de gestação foi confinado em instalações com piso de concreto. O piso era lavado três vezes por semana e as fezes eram removidas diariamente. O nascimento dos cordeiros ocorreu em Agosto e Setembro / 2001. Amostras de fezes foram colhidas das ovelhas e dos cordeiros no dia do nascimento, 4, 8, 16, 32 e 64 dias pós-parto. As amostras foram processadas pela técnica de centrífugosedimentação em éter. Esfregaços foram confeccionados e corados com auramina O e pela técnica de ZiehI-Neelsen modificada. Do total de amostras de cordeiros e ovelhas, 26,7% e 31,9%, respectivamente (P>O,O5) apresentaram oocistos de Crypostosporidium. Quatro dos 20 cordeiros e duas das 20 ovelhas não apresentaram oocistos em nenhuma das amostras examinadas. O percentual mais alto de amostras positivas ocorreu nas amostras dos cordeiros com quatro dias de vida. Nas ovelhas o maior percentual de amostras positivas foi registrado quatro dias após o parto. Apesar da taxa relativamente elevada de animais que eliminaram oocistos nas fezes, a infecção por C. parvum foi subclínica nas ovelhas e nos cordeiros.(AU)

Cryptosporidium parvum in captive primates of Parque Municipal Danilo Galafassi, Paraná, Brazil / Cryptosporidium parvum em primatas cativos do Parque Municipal Danilo Galafassi, Paraná, Brasil

A wide variety of terrestrial and aquatic animal species have been identified as hosts of species and genotypes of Cryptosporidium spp., which are important pathogens, however, little is known about their distribution in wild populations. Recent studies associating parasitological findings and molecular techniques have provided a new insight into host specificity and its potential transmission to humans.The objective of this study was to investigate the presence of Cryptosporidium spp. in feces of Callithrixsp. and Ateles paniscus, identify the species, and evaluate their phylogenetic relationships with other representatives of the genus. Four samples of feces were collected from an enclosure where three Callithrix jacchus and one Callithrix penicillate live; in addition, five samples were collected from an enclosure of an Ateles paniscus from Parque Municipal Danilo Galafassi, located in the city of Cascavel-PR. These samples were sent to the UFPR Biotechnology Laboratory, where the modified Ziehl-Neelsen staining technique was performed on microscope slides with fecal smear. Positive samples were submitted to DNA purification, extraction, PCR, and sequencing of the nuclear SSU rRNA region. Phylogenetic analysis based on Maximum Parcimony and Bayesian Inference were performed. Fifty percent (2: 4) of the feces samples from the enclosure of the Callithrix spp. and 60 % (3: 5) of samples from the Ateles paniscus enclosure were positive for Cryptosporidium spp. The phylogenetic analysis showed that the parasite found in both species of primates was recovered nested with others genotypes of C. parvum, and the genotype found in Callithrix spp. has high similarity with that one founded in several domestic animals. This is the first report of C. parvum in A. paniscus. Because it is an important zoonosis which does not have treatment, preventive measures must be adopted to avoid the spread of the disease.

Uma grande variedade de espécies animais terrestres e aquáticas tem sido identificada como hospedeiros de espécies e genótipos de Cryptosporidium spp., que são importantes agentes patogênicos, mas pouco se conhece sobre a sua distribuição nas populações silvestres. Estudos recentes associando achados parasitológicos e técnicas moleculares têm proporcionado uma nova visão em relação à especificidade do hospedeiro e seu potencial de transmissão para o homem. O objetivo desse estudo foi pesquisar a presença de Cryptosporidium spp. em fezes de Callithrix spp. e Ateles paniscus, identificar a espécie e avaliar o seu relacionamento filogenético com outros representantes do gênero. Foram coletadas quatro amostras de fezes de um recinto onde convivem três Callithrix jacchus e um Callithrix penicillata e cinco amostras de um recinto onde vive um Ateles paniscus do Parque Municipal Danilo Galafassi localizado na cidade de Cascavel-PR. As amostras foram enviadas ao Laboratório de Biotecnologia da UFPR onde foi realizada a técnica de coloração de Ziehl-Neelsen modificado em lâminas de esfregaço de fezes. As amostras positivas foram submetidas à purificação, extração de DNA, PCR, e sequenciamento da região nuclear SSU rRNA. Foram realizadas análises filogenéticas baseadas em Máxima Parcimônia e Inferência Bayesiana. Cinquenta por cento (2:4) das amostras de fezes do recinto dos Callithrix...

Avaliação da eficácia do colostro bovino hiperimune na infecção experimental de roedores com Cryptosporidium parvum / Evaluation of the efficacy of hyperimmune bovine colostrum on Cryptosporidium parvum experimental infection of rodents

O colostro bovino hiperimune contém alta concentração de anticorpos anti- Cryptosporidium parvum. Este produto é considerado como uma das estratégias terapêuticas promissoras no controle da criptosporidiose intestinal em humanos. Com o objetivo de avaliar a eficácia do colostro bovino hiperimune na redução do parasitismo e na prevenção de alterações de mucosa intestinal, foram infectados, experimentalmente, com oócistos de Cryptosporidium parvum, vários ratos (F344) e camundongos (C57BL/6). Os resultados mostraram que o colostro bovino hiperimune apresentou altos títulos de anticorpos. Os ratos F344, tratados com colostro bovino hiperimune ou com colostro normal, tiveram redução no parasitismo intestinal e apresentaram menor comprometimento de sua mucosa. Os camundongos C57BL/6, quando tratados com colostro hiperimune ou normal, apresentaram ligeira redução do parasitismo intestinal e não evidenciaram diferenças estatísticas significantes nas alterações histopatológicas da mucosa. Conclui-se que o uso do colostro bovino hiperimune tenha um benefício limitado na infecção causada pelo Cryptosporidium parvum.(AU)

The hyperimmune bovine colostrum has a high concentration of antibodies anti-Cryptosporidium parvum. This product is considered one of the promising therapeutic strategies in the control of intestinal cryptosporidiosis in humans. With the purpose to evaluate the hyperimmune bovine colostrum efficacy in reducing the parasitism and preventing intestinal mucosa alterations, several strains of rats F344 and mice were experimentally infected with Cryptosporidium parvum oocysts. The results showed that the hyperimmune bovine colostrum had high levels of antibodies. The rats F344, treated either with hyperimmune or normal bovine colostrum, had reduction of the intestinal parasitism and presented little mucosa compromise. The mice C57BL/6 when treated either with hyperimmune or normal colostrum had slight reduction of the intestinal parasitism and evidenced no statistical significant differences in the histopathological mucosa changes. In conclusion, the use of hyperimmune bovine colostrum has a limited benefit in Cryptosporidium parvum infection.(AU)

Cryptosporidium spp. em alimentos / Cryptosporidium spp. in food

O Cryptosporidium parvum foi descrito pela primeira vez em 1907 por Tyzzer, porém reconhecido como patógeno humano apenas em 1976. A Cryptosporidiose, causada principalmente pelo Cryptosporidium parvum, é mais grave em crianças, idosos e indivíduos imunodeficientes ou imunossuprimidos. Em contrapartida, em imunocompetentes, a doença é auto-limitante, caracterizada por uma diarréia aquosa que pode durar de 3 a 14 dias sem maiores complicações. O Cryptosporidium parvum é transmitido por oocistos, por meio de contato direto com as fezes de pessoas e animais infectados ou indiretamente durante a ingestão de água e alimentos contaminados. Os oocistos do Cryptosporidium spp. São resistentes aos desinfetantes químicos usados no tratamento da água de bebida, são facilmente destruídos pela fervura, mas não são retidos pelos filtros domésticos convencionais; porém, os filtros com poros com menos de 1 micrômetro de diâmetro e de osmose reversa são eficazes na retenção dos oocistos presentes na água. Os esgotos lançados (in natura) poluem os recursos hídricos, principalmente na falta de sistemas de captação e tratamento adequados. Estudos recentes demonstram que nos USA, 65-97 por cento das águas de superfície (rios, lagos, etc) estão contaminados com oocistos de Cryptosporidium spp, o que representa um sério risco ao aparecimento de surto de cryptosporidiose por meio da contaminação do sistema de abastecimento de água potável e da contaminação de frutas e verduras que são consumidas sem receber tratamento adequado. O surto de cryptosporidiose humana com maior repercussão, ocorreu na cidade de Milwaukee - Wisconsin - USA, envolvendo cerca de 300 mil pessoas, com um custo total de 90,2 milhões de dolares gastos no tratamento e controle da doença. O presente trabalho de revisão tem como objetivo principal alertar as autoridades e órgãos competentes para um maior controle microbiológico da água de abastecimento público e dos alimentos.(AU)

Cryptosporidium parvum was first described in 1907 by Tyzzer, however only recognized as a human pathogen in 1976. Cryptosporidiosis, caused primarily by Cryptosporidium parvum, is more severe in children, elderly and immune deficient or immune suppressed individuals. However, in immune competent individuals the disease is auto limited, characterized by liquid diarrhea that can last from 3 to 14 days without major complications. The Cryptosporidium parvum is transmitted by oocysts, either directly by contact with infected human or animal feces or indirectly by the ingestion of contaminated water or food. The Cryptosporidium spp. oocysts are resistant to chemical disinfectants used to treat municipal water supplies. They are easily destroyed by boiling water. C. parvum oocysts are not retained by conventional filtration systems. Special systems with pore size of less than 1 micrometer in diameter and with reverse osmosis systems are required to remove the infectious oocysts. Inadequate investment in sewage treatment systems leads to raw sewage polluting surface water supplies.(...)(AU)