Ocorrência da infecção por Cryptosporidium spp. em cabritos (Capra hircus) / Occurrence of infection by Cryptosporidium spp. in goat kids (Capra hircus)

O presente estudo teve como objetivo determinar a ocorrência da infecção por Cryptosporidium spp. em cabritos de Quixadá, Ceará, Brasil. Participaram do estudo 400 cabritos, com idade entre três e 360 dias, de ambos os sexos, com e sem padrão racial definido, procedentes de 25 estabelecimentos rurais distribuídos em três circuitos. As fezes foram cadastradas de acordo com o aspecto e cor, distribuídas em tubos tipo "eppendorf®" e congeladas in natura a -20°C, até o momento das extrações de DNA genômico do parasito com auxílio de kit comercial. Para amplificação de fragmentos da subunidade 18S do RNA ribossômico (rRNA) foi utilizada a "Nested"-PCR. A ocorrência de Cryptosporidium spp em cabritos de Quixadá foi de 7,50% (30/400). A frequência no período seco e no chuvoso foi de 9,55% (19/199) e 5,47% (11/201), respectivamente (χ²=2,39 e P>0,05). Amostras positivas foram identificadas em 64,00% (16/25) das propriedades estudadas e dessas amostras 50,00% (15/30) e 70,00% (21/30) tinham as fezes com aspecto e cor normais, respectivamente, sugerindo que cabritos assintomáticos estão eliminando oocistos. Não foi observada positividade para Cryptosporidium spp. em animais com 301 a 360 dias, demonstrando que animais mais velhos apresentam menos possibilidade de se infectarem com o parasito...

The present study aimed to determine the occurrence of infection by Cryptosporidium spp. in goat kids from Quixadá, Ceará, Brazil. The study included 400 goat kids of both sexes, 3 to 360 days old, with or without defined breed, originating from 25 farms distributed in three circuits. Feces were registered in accordance with the appearance and color, distributed into tubes Eppendorf tubes and frozen in natura at-20°C until the moment of extraction of genomic DNA from the parasite with the aid of a commercial kit. For amplification of fragments of the 18S subunit of ribosomal RNA (rRNA) was used to Nested PCR. The occurrence of Cryptosporidium spp. in goats kids of the Quixadá was 7.50% (30/400). The frequency in the dry period and rainy was 9.55% (19/199) and 5.47% (11/201) respectively (χ²=2.39 and P>0.05). Positive samples were identified in 64.00% (16/25) of the studied farms, and from these samples 50.00% (15/30) and 70.00% (21/30) had feces with normal appearance and color respectively, suggesting that the asymptomatic goats were eliminating oocysts. No positivity for Cryptosporidium spp. was observed in 301 to 360-day-old goats, demonstrating that older animals have less chance to become infected with the parasite...

Surto de criptosporidiose em bezerros no Sul do Rio Grande do Sul / Outbreak of cryptosporidiosis in calves in Southern Brazil

Descrevem-se os aspectos epidemiológicos, sinais clínicos e a patologia de um surto de criptosporidiose em bezerros na região Sul do Rio Grande do Sul. De um lote de 400 bezerros de 30-45 dias de idade, 35 adoeceram e 16 morreram. Os bezerros nasciam fracos e logo após o nascimento apresentavam diarreia amarela, emagrecimento progressivo, desidratação, depressão e morte entre 10 e 15 dias após o início dos sinais clínicos. Na necropsia havia congestão dos vasos sanguíneos intestinais e mesentéricos. Havia distensão intestinal por gás e dilatação de vasos linfáticos. Microscopicamente havia achatamento das vilosidades intestinais, com necrose e atrofia. Aderidas à superfície das células epiteliais das vilosidades, havia estruturas puntiformes basofílicas de 2-5µm de diâmetro compatíveis com Cryptosporidium spp. A microscopia eletrônica revelou a presença de diferentes estágios do agente aderidos às microvilosidades de enterócitos. Alerta-se para a importância da criptosporidiose como agente primário de diarreia em bezerros. São necessárias medidas preventivas no que se refere ao manejo para diminuir as perdas econômicas e a contaminação ambiental, e, ainda, diminuir o risco para a saúde pública...

This paper describes the epidemiology, clinical signs and pathology of an outbreak of cryptosporidiosis in calves in Southern Brazil. Thirty-five out of 400 calves with 30-45 days of age were affected and 16 died. The calves were born weak and just after birth they had yellow diarrhea, weight loss, dehydration, depression, and death between 10 and 15 days after onset of clinical signs. Congestion of the bowel and mesenteric blood vessels were observed at necropsy. Intestinal distension by gas and dilation of lymphatic vessels were also observed. Microscopically, the intestine showed flattening of the villi with necrosis and atrophy. Adhered to the surface of the villus epithelial cells there were round basophilic structures of 2- to 5-μm-diameter compatible with Cryptosporidium spp. Electron microscopy revealed the presence of different stages of the agent adhered to the microvilli of enterocytes. We alert the importance of cryptosporidiosis as a primary agent of diarrhea in calves. Preventive measures to reduce economic losses, environmental contamination, and also decrease of risk for public health are necessary...

Ocorrência da infecção por Cryptosporidium spp. em cabritos (Capra hircus) / Occurrence of infection by Cryptosporidium spp. in goat kids (Capra hircus)

O presente estudo teve como objetivo determinar a ocorrência da infecção por Cryptosporidium spp. em cabritos de Quixadá, Ceará, Brasil. Participaram do estudo 400 cabritos, com idade entre três e 360 dias, de ambos os sexos, com e sem padrão racial definido, procedentes de 25 estabelecimentos rurais distribuídos em três circuitos. As fezes foram cadastradas de acordo com o aspecto e cor, distribuídas em tubos tipo "eppendorf®" e congeladas in natura a -20°C, até o momento das extrações de DNA genômico do parasito com auxílio de kit comercial. Para amplificação de fragmentos da subunidade 18S do RNA ribossômico (rRNA) foi utilizada a "Nested"-PCR. A ocorrência de Cryptosporidium spp em cabritos de Quixadá foi de 7,50% (30/400). A frequência no período seco e no chuvoso foi de 9,55% (19/199) e 5,47% (11/201), respectivamente (χ²=2,39 e P>0,05). Amostras positivas foram identificadas em 64,00% (16/25) das propriedades estudadas e dessas amostras 50,00% (15/30) e 70,00% (21/30) tinham as fezes com aspecto e cor normais, respectivamente, sugerindo que cabritos assintomáticos estão eliminando oocistos. Não foi observada positividade para Cryptosporidium spp. em animais com 301 a 360 dias, demonstrando que animais mais velhos apresentam menos possibilidade de se infectarem com o parasito.(AU)

The present study aimed to determine the occurrence of infection by Cryptosporidium spp. in goat kids from Quixadá, Ceará, Brazil. The study included 400 goat kids of both sexes, 3 to 360 days old, with or without defined breed, originating from 25 farms distributed in three circuits. Feces were registered in accordance with the appearance and color, distributed into tubes Eppendorf tubes and frozen in natura at-20°C until the moment of extraction of genomic DNA from the parasite with the aid of a commercial kit. For amplification of fragments of the 18S subunit of ribosomal RNA (rRNA) was used to Nested PCR. The occurrence of Cryptosporidium spp. in goats kids of the Quixadá was 7.50% (30/400). The frequency in the dry period and rainy was 9.55% (19/199) and 5.47% (11/201) respectively (χ²=2.39 and P>0.05). Positive samples were identified in 64.00% (16/25) of the studied farms, and from these samples 50.00% (15/30) and 70.00% (21/30) had feces with normal appearance and color respectively, suggesting that the asymptomatic goats were eliminating oocysts. No positivity for Cryptosporidium spp. was observed in 301 to 360-day-old goats, demonstrating that older animals have less chance to become infected with the parasite.(AU)

Surto de criptosporidiose em bezerros no Sul do Rio Grande do Sul / Outbreak of cryptosporidiosis in calves in Southern Brazil

Descrevem-se os aspectos epidemiológicos, sinais clínicos e a patologia de um surto de criptosporidiose em bezerros na região Sul do Rio Grande do Sul. De um lote de 400 bezerros de 30-45 dias de idade, 35 adoeceram e 16 morreram. Os bezerros nasciam fracos e logo após o nascimento apresentavam diarreia amarela, emagrecimento progressivo, desidratação, depressão e morte entre 10 e 15 dias após o início dos sinais clínicos. Na necropsia havia congestão dos vasos sanguíneos intestinais e mesentéricos. Havia distensão intestinal por gás e dilatação de vasos linfáticos. Microscopicamente havia achatamento das vilosidades intestinais, com necrose e atrofia. Aderidas à superfície das células epiteliais das vilosidades, havia estruturas puntiformes basofílicas de 2-5µm de diâmetro compatíveis com Cryptosporidium spp. A microscopia eletrônica revelou a presença de diferentes estágios do agente aderidos às microvilosidades de enterócitos. Alerta-se para a importância da criptosporidiose como agente primário de diarreia em bezerros. São necessárias medidas preventivas no que se refere ao manejo para diminuir as perdas econômicas e a contaminação ambiental, e, ainda, diminuir o risco para a saúde pública.(AU)

This paper describes the epidemiology, clinical signs and pathology of an outbreak of cryptosporidiosis in calves in Southern Brazil. Thirty-five out of 400 calves with 30-45 days of age were affected and 16 died. The calves were born weak and just after birth they had yellow diarrhea, weight loss, dehydration, depression, and death between 10 and 15 days after onset of clinical signs. Congestion of the bowel and mesenteric blood vessels were observed at necropsy. Intestinal distension by gas and dilation of lymphatic vessels were also observed. Microscopically, the intestine showed flattening of the villi with necrosis and atrophy. Adhered to the surface of the villus epithelial cells there were round basophilic structures of 2- to 5-μm-diameter compatible with Cryptosporidium spp. Electron microscopy revealed the presence of different stages of the agent adhered to the microvilli of enterocytes. We alert the importance of cryptosporidiosis as a primary agent of diarrhea in calves. Preventive measures to reduce economic losses, environmental contamination, and also decrease of risk for public health are necessary.(AU)

High occurrence of Cryptosporidium ubiquitum and Giardia duodenalis genotype E in sheep from Brazil.

In this paper we report the occurrence of zoonotic species of Cryptosporidium (C. ubiquitum and C. parvum subtype IIaA15G2R1) and the host-specific Giardia duodenalis genotype E in a sheep farm from Brazil. Fecal specimens were collected from 100 animals and screened for the presence of G. duodenalis and Cryptosporidium spp. by enzyme immunoassay (EIA) and polymerase chain reaction (PCR). Restriction fragment length polymorphism (RFLP) and sequence analysis of selected samples was used to confirm the species and to characterize the isolates. Of 100 specimens, 34 were positive for Giardia (34%) and 25 were positive for Cryptosporidium (25%). The prevalence of Giardia and Cryptosporidium was higher in lambs than in ewes. Co-infection was observed in 14% of animals. All G. duodenalis isolates belonged to genotype E. The genetic characterization performed in 22 Cryptosporidium-positive samples revealed the presence of zoonotic C. ubiquitum and C. parvum in 76% (19/25) and 12% (3/25) of infected animals, respectively. The presence of zoonotic Cryptosporidium subtypes in lambs was of epidemiological and public health relevance.

Molecular characterization of Cryptosporidium isolates from calves in Argentina.

Cryptosporidiosis is responsible for significant fatalities of neonatal calves, resulting in substantial economic loss in dairy farming in several countries. Additionally, the high shedding of environmentally resistant oocysts by calves promotes contamination of drinking water and facilitates outbreaks of cryptosporidiosis in humans. Here we report on the Cryptosporidium species and GP60 subtypes of 45 calves originating from the Humid Pampa, the main productive dairy farming area of Argentina. Polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the 18S rRNA gene was done to determine the infecting Cryptosporidium species and only Cryptosporidium parvum was detected. Subtyping by sequence analysis of the GP60 gene revealed 6 different alleles all pertaining to the zoonotic IIa family. Of these, IIaA23G1R1 represents a novel IIa subtype. Other identified subtypes, IIa18G1R1, IIaA20G1R1, IIaA21G1R1, and IIaA22G1R1 have been recognized in very few studies and/or with low frequencies. Interestingly, different alleles prevailed in the provinces of Buenos Aires (IIaA17G1R1 and IIaA21G1R1), Santa Fe (IIaA23G1R1), and Cordoba (IIaA20G1R1 and IIaA21G1R1), and different allele distribution patterns were observed. Subtypes IIaA18G1R1 and IIaA17G1R1, the latter often found in this study, are strongly implicated in zoonotic transmission, suggesting that calves may represent a potential source for human cryptosporidiosis in this region. This is the first published report of a molecular analysis of Cryptosporidium infection in dairy and beef calves from Argentina.

Clinical, serological, and parasitological analysis of snakes naturally infected with Cryptosporidium serpentis.

Infection by Cryptosporidium serpentis is one of the most important diseases in reptiles and is characterized by chronic clinical or subclinical infection and the presence of hypertrophic gastritis, food regurgitation, progressive weight loss, mortality, and intermittent or continuous shedding of oocysts in the feces. The objectives of this study were to standardize an indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies against C. serpentis and to evaluate the clinical, parasitological, and humoral immune response in snakes naturally infected with C. serpentis. Twenty-one snakes naturally infected with C. serpentis and housed at the Butantan Institute, São Paulo, Brazil, underwent clinical and parasitological analyses for C. serpentis infection through daily records of clinical signs and a monthly survey of fecal shedding of oocysts using the Kinyoun's acid-fast staining. The serological evaluation was performed monthly by indirect ELISA using crude total antigen from oocysts of C. serpentis to detect anti-C. serpentis antibodies. Clinical symptoms consisted of food regurgitation, inappetence, and progressive weight loss. The parasitological analysis revealed intermittent fecal shedding of a variable number of oocysts in all snakes, with positivity in 85.32% (157/184) of the samples. The indirect ELISA was positive in 68.25% (86/126) of the samples. A humoral immune response was observed in most animals; however, fluctuating antibodies levels, leading to alternating positive and negative results, were observed in most snakes.

A new set of primers directed to 18S rRNA gene for molecular identification of Cryptosporidium spp. and their performance in the detection and differentiation of oocysts shed by synanthropic rodents.

Cryptosporidium spp. are cosmopolitan protozoa that infect fishes, reptiles, amphibians, birds and mammals. More than 20 species are recognized within this genus. Rodents are a group of abundant and ubiquitous organisms that have been considered reservoirs of Cryptosporidium for humans and livestock. The aim of this study was to design specific primers for the gene encoding 18S rRNA, potentially capable of amplifying any species or genotype of Cryptosporidium spp. and evaluate the diagnostic attributes of the nested-PCR based on such probes. The primers were designed to amplify the shortest segment as possible to maximize the sensitivity of the test, but preserving the discriminatory potential of the amplified sequences for phylogenetic inferences. The nested-PCR standardized in this study (nPCR-SH) was compared in terms of sensitivity with another similar assay (nPCR-XIAO) that has been largely used for the detection and identification of Cryptosporidium spp. worldwide. We also aimed to molecularly characterize samples of Cryptosporidum spp. isolated from synanthropic rodents using these probes. Forty-five rodents were captured in urban areas of the municipality of Umuarama, Paraná State, Brazil. Fecal samples were submitted to three molecular tests (nested-PCRs), two of them targeted to the 18S rDNA gene (nPCR-SH and nPCR-XIAO) and the third targeted to the gene encoding actin (nPCR-actin). The nPCR-SH was tested positive on samples of Cryptosporidum parvum, Cryptosporidum andersoni, Cryptosporidum meleagridis, Cryptosporidum hominis, Cryptosporidum canis, and Cryptosporidum serpentis. Sixteen samples of rodents were positive by nPCR-SH, six by nPCR-XIAO and five by nPCR-actin. Sequencing of amplified fragments allowed the identification of Cryptosporidum muris in three samples of Rattus rattus, and two genotypes of Cryptosporidium, the genotypes mouse II and III. Cryptosporidium genotype mouse II was found in one sample of Mus musculus and genotype mouse III, in twelve samples, being five from R. rattus and seven from M. musculus. The results of this study demonstrated that the primers designed for detection of Cryptosporidium spp. were more efficient than those used in the nPCR-XIAO. Genotypes or species of Cryptosporidium that can be usually transmitted for human beings and livestock were not found in synanthropic rodents, suggesting that the importance of these animals in zoonotic transmission of cryptosporidiosis should be revisited.

[Cryptosporidium spp. and Giardia spp. prevalence and risk factors in dairy calves of the north-western zone of the Bogota Savanna]. / Prevalencia y factores de riesgo de Cryptosporidium spp. y Giardia spp. en terneros de ganado lechero de la zona noroccidental de la Sabana de Bogotá

OBJECTIVE: The present study was aimed to establishing Cryptosporidium spp. and Giardia spp. prevalence in 0 to 2 months old dairy calves of the north-western zone of the Bogota Savanna. In addition, associated factors related to a failure in Good Practices of Livestock could incur in human and animal infection. METHODS: This was a cross-sectional study; calves' fecal samples were used. Farms' Good Practices of Livestock were observed by means of an observation blank. Giardia spp. and Cryptosporidium spp. prevalence was determinate by means of laboratory results (Ritchie for Giardia spp. and modified Ziehl-Neelsen for Cryptosporidium spp.). Odds Ratios (OR) were calculated in association between this two genera of protozoa and Good Practices of Livestock. RESULTS: Thirty three dairy farms were evaluated, where fecal samples of 308 calves were taken. Giardia spp. prevalence was 37.7 %, 115 infected animals; Cryptosporidium spp. prevalence was 4.9 %, 15 infected animals. CONCLUSION: There is an important Giardia spp. and Cryptosporidium foci in the north-western zone of the Bogota Savanna, without a previous knowledge. Giardia spp. prevalence for this zone is in the highest rank reported for South-America and Cryptosporidium spp. prevalence is in en the lowest one. Associated risk factors of Giardia spp. and Cryptosporidium spp. in dairy farms of the north-western zone of the Bogota Savanna depend of a Good Practices of Livestock performance.

Cryptosporidium spp. infection in mares and foals of the northwest region of São Paulo State, Brazil.

The present study aimed to analyze the occurrence of infection by Cryptosporidium spp. in mares and their respective foals. This study was carried out in 11 farms located in the municipalities of Araçatuba, Birigui, Guararapes and Santo Antônio do Aracangua, in the northwest region of the State of Sao Paulo, from November 2010 to March 2011. A total of 98 mares and 98 foals of several breeds were analyzed; among foals, 59 were males and 39 females, aged from three to 330 days. Feces were collected directly from the rectal ampulla, purified and processed according to modified Kinyoun stain. Occurrence of Cryptosporidium spp. was 21.4% (21/98) for foals and 18.4% (18/98) for mares. Occurrence of Cryptosporidium spp. had significant association with breeds and age of animals. Results obtained led to the conclusion that foals older than two months and Mangalarga animals are less susceptible to the occurrence of Cryptosporidium spp.

Cryptosporidium em bezerros destinados a exploração leiteira no estado do rio de janeiro: padronização das técnicas moleculares, caracterização genotípica e influência do manejo na infecção

A criptosporidiose bovina é de grande relevância para a pecuária brasileira, principalmente em animais jovens. Bovinos infectados com Cryptosporidium podem apresentar um baixo desempenho produtivo além de eliminar grande quantidade de formas infectantes no ambiente, especialmente aqueles que apresentam um quadro clínico de diarréia. Os objetivos do trabalho foram: comparar duas técnicas de extração de DNA utilizando um kit comercial e um protocolo adaptado por pesquisadores; analisar diversas temperaturas de anelamento para as reações de PCR-Nested para os genes alvo 18S e GP60, além de dois tipos de coloração de fragmentos de DNA em gel de agarose (brometo de etídio e gelred); determinar a ocorrência de espécies e subtipos de Cryptosporidium em bezerros com até um ano de idade, através da técnica de PCR para os genes alvos 18S e GP60 e analisar a influencia de modificações no manejo no percentual de infecção em bezerros. Os resultados da padronização demostraram uma maior grau de pureza do kit comercial, tendo sido ainda estabelecido novas temperaturas para uma otimização das técnicas, além da maior sensibilidade e baixa toxicidade do corante gelred. Após o sequenciamento foi observada a presença de três espécies de Cryptosporidium infectando bezerros em diferentes faixas etárias. Na fase pré-desmame (< 2 meses) 10% dos animais encontravam-se infectados por C. parvum, enquanto que os bezerros na fase pósdesmame (? 2 meses) apresentaram-se infectados por C. andersoni (15%) e C. ryanae (2%). Nesse estudo também foi observado que a implantação de algumas mudanças no manejo da propriedade pode ser de grande ajuda na tentativa de diminuir a disseminação da doença entre os animais e reduzir a contaminação ambiental

Cryptosporidium species screening using Kinyoun technique in domestic cats with diarrhea.

Cryptosporidium is a coccidian that can lead to diarrhea, especially in immunosuppressed individuals. Retroviruses are considered a primary cause of immunosuppression in cats. Fecal specimens and blood collected from the 60 cats were evaluated for the presence of acid-fast cryptosporidia in three consecutive stool samples and for feline leukemia virus (FeLV) antigen and feline immunodeficiency virus (FIV) antibody by ELISA testing. Five animals (8.33%) shedding oocysts were found, one was both FIV- and FeLV-negative and four were FeLV-positive.

Presence and molecular characterisation of Giardia and Cryptosporidium in alpacas (Vicugna pacos) from Peru.

The presence of Giardia and Cryptosporidium was investigated in 274 faecal samples of alpacas (Vicugna pacos) from 12 herds from Peru by immunofluorescence microscopy and PCR amplification and sequencing of fragments of the ssu-rRNA and ß-giardin genes from Giardia spp., as well as the ssu-rRNA gene from Cryptosporidium spp. A total of 137 samples (50.0%) were positive for Giardia spp., and 12 samples (4.4%) for Cryptosporidium spp. In ten samples (3.6%), co-infection by both pathogens was found. Herd prevalence was found to be 91.7% (11/12 herds) for Giardia and 58.3% (7/12 herds) for Cryptosporidium. Regarding the age of the animals, although Giardia was detected in animals as young as 1 week, the prevalence increased with age, reaching 80% by 8 weeks. Similarly, the highest percentage of Cryptosporidium detection (20%) was also found in the 8 week-old group. By PCR, 92 of the 274 analysed samples were positive for Giardia. Sequencing of the amplicons showed the existence of Giardia duodenalis assemblage A in 67 samples; G. duodenalis assemblage E in 24 samples; and inconsistent results between the two molecular markers used in a further sample. Cryptosporidium was only detected by PCR in 3 of the 274 samples; Cryptosporidium parvum was identified in two samples and Cryptosporidium ubiquitum in one sample. This study is the first performing molecular characterisation of both parasites in Peruvian alpacas, and the first report of C. ubiquitum in this host. The identification of G. duodenalis assemblage A, C. parvum and C. ubiquitum, suggests that zoonotic transmission of these enteropathogens between alpacas and humans is possible.

Real-time PCR assay targeting the actin gene for the detection of Cryptosporidium parvum in calf fecal samples.

Cryptosporidium parvum infection is very important with respect to public health, owing to foodborne and waterborne outbreaks and gastrointestinal illness in immunocompetent and immunocompromised persons. In cattle, infection with this species manifests either as a subclinical disease or with diarrheal illness, which occurs more often in the presence of other infectious agents than when alone. The aim of this study was to develop a real-time polymerase chain reaction (PCR) assay for the detection of C. parvum in calf fecal samples and to compare the results of this assay with those of the method routinely used for the diagnosis of Cryptosporidium spp., nested PCR targeting the 18S rRNA gene. Two hundred and nine fecal samples from calves ranging in age from 1 day to 6 months were examined using real-time PCR specific for the actin gene of C. parvum and by a nested PCR targeting the 18S rRNA gene of Cryptosporidium spp. Using real-time PCR detection, 73.2% (153 out of 209) of the samples were positive for C. parvum, while 56.5% (118 out of 209) of the samples were positive for Cryptosporidium spp. when the nested PCR amplification method was used for the detection. The analytical sensitivity of the real-time PCR was approximately one C. parvum oocyst. There was no significant nonspecific DNA amplification of any of the following species and genotype: Cryptosporidium andersoni, Cryptosporidium baileyi, Cryptosporidium bovis, Cryptosporidium canis, Cryptosporidium galli, Cryptosporidium ryanae, Cryptosporidium serpentis, or avian genotype II. Thus, we conclude that real-time PCR targeting the actin gene is a sensitive and specific method for the detection of C. parvum in calf fecal samples.

Cryptosporidium spp. parasitize exotic birds that are commercialized in markets, commercial aviaries, and pet shops.

The purpose of this study was to genetically characterize and phylogenetically analyze the Cryptosporidium spp. isolated from exotic birds commercialized in popular markets, commercial aviaries, and pet shops located in Rio de Janeiro, Brazil. Fecal samples from individually housed birds were collected and subjected to centrifuge-flotation technique using saturated sugar solution. DNA was isolated from Cryptosporidium positive samples, and 18S subunit rDNA was amplified and processed using nested-polymerase chain reaction (PCR). To identify the protozoan species, the PCR amplicons were used for restriction fragment length polymorphism and sequencing analyses. Of the 103 analyzed fecal samples, seven (6.8%) were positive for Cryptosporidium oocysts. Sequencing and further phylogenetic analyses allowed us to identify the following species: Cryptosporidium parvum in Bengalese finch (Lonchura striata domestica) and avian genotype III in Java sparrow (Padda oryzivora) and cockatiel (Nymphicus hollandicus). The sequences of the Cryptosporidium spp. isolated from canaries (Serinus canarius) were not identifiable within the groups of known species, but they presented a higher genetic similarity with C. parvum. This is the first report in Brazil showing that C. parvum parasitizes Bengalese finches and that avian genotype III parasitizes Java sparrows.

Prevalencia y factores de riesgo de Cryptosporidium spp. y Giardia spp. en terneros de ganado lechero de la zona noroccidental de la Sabana de Bogotá / Cryptosporidium spp. and Giardia spp. prevalence and risk factors in dairy calves of the north-western zone of the Bogota Savanna

Objetivo Estimar la prevalencia de Cryptosporidium spp. y Giardia spp. en terneros de 0 a 2 meses de ganado lechero de la zona noroccidental de la Sabana de Bogotá. Se estimaron los factores de riesgo de la producción lechera que puedan incurrir en la infección de personas y animales. Métodos Estudio de corte transversal, mediante la toma de materia fecal de terneros. Se determinó la prevalencia de punto para Giardia spp. y Cryptosporidium spp. a través los resultados de los coprológicos (Ritchie para Giardia spp. y Ziehl-Neelsen modificada para Cryptosporidium spp.). Se calcularon Odds Ratio para establecer los factores de riesgo asociados entre estos dos géneros de protozoarios y el manejo de las Buenas Prácticas Ganaderas. Resultados Se evaluaron 33 fincas dedicadas a la producción lechera, donde se muestrearon 308 terneros. La prevalencia para Giardia spp. fue de 37,3 %, 115 animales positivos y para Cryptosporidium spp. fue de 4,9 % 15, animales positivos. Conclusión Existe un foco de Giardia spp. y Cryptosporidium spp. en el noroccidente de la Sabana de Bogotá, del cual no se tenía conocimiento previo. La prevalencia de Giardia spp. para la región estudiada, está en el límite alto del rango reportado para Sudamérica. En el caso de Cryptosporidium spp. la situación es distinta, la prevalencia se encuentra en el limite bajo del rango reportado para Sudamérica. Los factores de riesgo asociados a Giardia spp. y Cryptosporidium spp. en las explotaciones lecheras del noroccidente de la Sabana de Bogotá, dependen de Buenas Prácticas Ganaderas.

Objective The present study was aimed to establishing Cryptosporidium spp. and Giardia spp. prevalence in 0 to 2 months old dairy calves of the north-western zone of the Bogota Savanna. In addition, associated factors related to a failure in Good Practices of Livestock could incur in human and animal infection. Methods This was a cross-sectional study; calves' fecal samples were used. Farms' Good Practices of Livestock were observed by means of an observation blank. Giardia spp. and Cryptosporidium spp. prevalence was determinate by means of laboratory results (Ritchie for Giardia spp. and modified Ziehl-Neelsen for Cryptosporidium spp.). Odds Ratios (OR) were calculated in association between this two genera of protozoa and Good Practices of Livestock. Results Thirty three dairy farms were evaluated, where fecal samples of 308 calves were taken. Giardia spp. prevalence was 37.7 %, 115 infected animals; Cryptosporidium spp. prevalence was 4.9 %, 15 infected animals. Conclusion There is an important Giardia spp. and Cryptosporidium foci in the north-western zone of the Bogota Savanna, without a previous knowledge. Giardia spp. prevalence for this zone is in the highest rank reported for South-America and Cryptosporidium spp. prevalence is in en the lowest one. Associated risk factors of Giardia spp. and Cryptosporidium spp. in dairy farms of the north-western zone of the Bogota Savanna depend of a Good Practices of Livestock performance.

Occurrence of Cryptosporidium spp. in Antillean manatees (Trichechus manatus) and Amazonian manatees (Trichechus inunguis) from Brazil.

Infections by Cryptosporidium spp. in aquatic mammals is a major concern due to the possibility of the waterborne transmission of oocysts. The aim of the present study was to report the occurrence of Cryptosporidium spp. in Antillean manatees (Trichechus manatus) and Amazonian manatees (Trichechus inunguis) from Brazil. Fecal samples were collected and processed using Kinyoun's method. Positive samples were also submitted to the direct immunofluorescence test. The results revealed the presence of Cryptosporidium spp. oocysts in 12.5% (17/136) of the material obtained from the Antillean manatees and in 4.3% (05/115) of the samples from the Amazonian manatees. Cryptosporidium spp. infection was more prevalent in captive animals than in free-ranging specimens.

Prevalence of Cryptosporidium spp. and Giardia spp., spatial clustering and patterns of shedding in dairy calves from Córdoba, Argentina.

The objectives of this study were to estimate calf and herd prevalence of Cryptosporidium spp. and Giardia spp., the herd prevalence clustering, spatial distribution according to soil type and shedding patterns in dairy calves from Cordoba, Argentina. Six hundred twenty calves younger than 7 weeks of age from 43 dairy herds were sampled. Samples were processed with the formol-ether and modified Ziehl-Neelsen techniques. Univariate analysis and Kruskall-Wallis tests were used. Factors associated were subjected to multivariate analysis with calf shedding intensity as the response variable. Clustering of herd prevalence was assessed by a scan method, and spatial analysis was applied to explore the overlapping of high prevalence herds and soil type. Overall calf prevalence for Cryptosporidium spp. oocysts and Giardia spp. cysts were 19.35% (95% CI: 16.14; 22.54) and 34.50% (95% CI: 30.69; 38.34), respectively. Calves younger than two weeks of age were almost four times more likely to be infected with Cryptosporidium, in comparison to older ones (RR: 3.78, 95% CI: 2.27; 6.26). Giardia spp. shedding showed a similar age pattern (RR: 1.33, 95% CI: 1.02; 1.75). A primary cluster of high Cryptosporidium prevalence was found, and high prevalence herds were located in areas with poor drained soil.

Prevalence of Cryptosporidium spp. and Giardia spp., spatial clustering and patterns of shedding in dairy calves from Córdoba, Argentina / Prevalência de Cryptosporidium spp. e Giardia spp., distribuição espacial e padrões de eliminação em bezerros de propriedades leiteiras em Córdoba, Argentina

The objectives of this study were to estimate calf and herd prevalence of Cryptosporidium spp. and Giardia spp., the herd prevalence clustering, spatial distribution according to soil type and shedding patterns in dairy calves from Cordoba, Argentina. Six hundred twenty calves younger than 7 weeks of age from 43 dairy herds were sampled. Samples were processed with the formol-ether and modified Ziehl-Neelsen techniques. Univariate analysis and Kruskall-Wallis tests were used. Factors associated were subjected to multivariate analysis with calf shedding intensity as the response variable. Clustering of herd prevalence was assessed by a scan method, and spatial analysis was applied to explore the overlapping of high prevalence herds and soil type. Overall calf prevalence for Cryptosporidium spp. oocysts and Giardia spp. cysts were 19.35 percent (95 percent CI: 16.14; 22.54) and 34.50 percent (95 percent CI: 30.69; 38.34), respectively. Calves younger than two weeks of age were almost four times more likely to be infected with Cryptosporidium, in comparison to older ones (RR: 3.78, 95 percent CI: 2.27; 6.26). Giardia spp. shedding showed a similar age pattern (RR: 1.33, 95 percent CI: 1.02; 1.75). A primary cluster of high Cryptosporidium prevalence was found, and high prevalence herds were located in areas with poor drained soil.

Os objetivos deste estudo foram determinar a prevalência de Cryptosporidium spp. e Giardia spp., a presença de aglomerados, a distribuição espacial de acordo com o tipo de solo e padrões de eliminação de cistos e oocistos em bezerros de propriedades leiteiras em Córdoba, Argentina. Amostras fecais foram colhidas de 620 bezerros com menos de sete semanas de idade, provenientes de 43 propriedades leiteiras e examinadas pelas técnicas de formol-éter e Ziehl-Neelsen modificada. Foram realizadas uma análise univariada e o teste de Kruskal-Wallis e, em seguida, uma análise multivariada com a intensidade de eliminação de cistos e oocistos, como um evento. A presença de aglomerados foi determinada com o método de varredura e a análise espacial foi realizada para explorar a sobreposição de rebanhos com alta prevalência e tipo de solo. A prevalência de Cryptosporidium spp. e Giardia spp. foi de 19,35 por cento (IC 95 por cento: 16,14; 22,54) e 34,50 por cento (IC 95 por cento: 30,69; 38,34), respectivamente. A probabilidade de infecção por Cryptosporidium spp. foi quase quatro vezes maior para bezerros com menos de 2 semanas em comparação com os bezerros mais velhos (RR: 3,78, IC 95 por cento 2,27; 6,26). O mesmo padrão de infecção relacionada à idade foi observado para Giardia spp. (RR: 1,33, IC 95 por cento 1,02; 1,75). Foi encontrado um aglomerado primário com alta prevalência de Cryptosporidium spp., e rebanhos com alta prevalência foram localizados em solos mal drenados.

Occurrence of Cryptosporidium andersoni in Brazilian cattle.

Feces were collected from 68 dairy cattle, 1 to 12 mo of age, on 12 farms in the municipality of Campos dos Goytacazes, Rio de Janeiro, Brazil, and examined for the presence of Cryptosporidium sp. All samples were subjected to molecular analysis by polymerase chain reaction (nested PCR) of the 18S rRNA. Four positive samples (4.54%) were sequenced and identified as Cryptosporidium andersoni. This species represents a risk for Brazilian cattle because infection can affect cattle productivity. Moreover, C. andersoni is considered a zoonotic species.