First report of Amblyomma sculptum (Amblyomma cajennense complex) in a Brazilian state classified as a silent area for human rickettsiosis.

Background and Aim: Studies on ticks of public health concern in equine husbandry are scarce in the Northeastern region of Brazil. This study aimed to investigate the presence of ticks on horses in the State of Alagoas, which is classified as a silent area for human rickettsiosis. Materials and Methods: Ticks infesting horses were collected using anatomical tweezers or a commercial hook and kept in ethanol-labeled tubes for taxonomic identification. Results: A total of 2,238 ticks were found. Ticks were identified as 2,215 (98.89%, 95% CI: 98.41-99.28) Dermacentor nitens, 19 (0.98%, 95% CI: 0.05-1.38) Amblyomma sculptum, and 4 (0.18%; 95% CI: 0.007-0.46) Rhipicephalus microplus. Conclusion: This is the first study to report A. sculptum and D. nitens in the State of Alagoas. The presence of A. sculptum should draw the attention of public health managers once Alagoas State is considered a silent area for rickettsial diseases, which means the absence of local surveillance programs for these pathogens.

Protection against Toxoplasma gondii cysts in pigs immunized with rROP2 plus Iscomatrix.

This study aimed to evaluate the humoral immune response in pigs immunized intranasally and intramuscularly with recombinant Toxoplasma gondii rROP2 protein in combination with the adjuvant Iscomatrix. Twelve mixed breed pigs divided into three groups (n=4) were used, G1 received recombinant ROP2 proteins (200 µg/dose) plus Iscomatrix, G2 received PBS plus Iscomatrix, and G3 as the control group. The intranasal (IN) and intramuscular (IM) routes were used. Animals were challenged orally with VEG strain oocysts and treated on day three after challenge. Fever, anorexia, and prostration were the clinical signs observed in all animals. All the G1 animals produced antibodies above the cut-off on the day of the challenge, while the G2 and G3 remained below the cut-off. Better partial protection against parasitemia and cyst tissue formation was observed in G1 than G3. The protection factors against tissue cyst formation were 40.0% and 6.1% for G1 and G2, respectively, compared to G3. In conclusion, there were not systemic antibody responses in pigs with IN immunization with rROP2+Iscomatrix; however, after IM immunization, those animals produced higher titers than animal controls. We associated these results with partial protection obtained against parasitemia and tissue cysts formation.

Neospora caninum DNA in feces of crab-eating fox (Cerdocyon thous - Linnaeus, 1776) from northeastern Brazil.

Neospora caninum is an obligate intracellular tissue cyst-forming coccidian parasite and it was first described in dogs. Despite the relevance of wild canids in the transmission chain of N. caninum, there are few studies in Brazil. The aim of the present study was to detect N. caninum DNA in feces of free-range and captive crab-eating fox (Cerdocyon thous) from different states of northeastern Brazil. Fecal samples of eighteen crab-eating foxes (fifteen individually and three pools) were collected in sterile containers and were kept cool at -20 °C until further processing. All fecal samples were subjected to DNA extraction. A nested PCR targeting the ITS-1 gene was performed for N. caninum. All the positive bands were extracted from the gel and purified. Forward and reverse strands were sequenced and the nucleotide sequences obtained were compared with N. caninum sequences deposited in Genbank. The alignment was edited and phylogenetic reconstruction was based on the ITS1 gene sequences. Thirteen stool samples were PCR-positive for N. caninum DNA. Nine out of thirteen positive samples showed similarity between 99%-100% for N. caninum in relation to the sequence U25044.1 stored at GenBank. The crab-eating fox could have an important role in the sylvatic cycle of Neospora caninum in Brazil. Experimental infections studies involving these wild canids may confirm if the crab-eating foxes are definitive hosts of N. caninum.

First description of clonal lineage type II (genotype #1) of Toxoplasma gondii in abortion outbreak in goats.

The purpose of this study was to perform genotypic characterization and to evaluate the virulence of Toxoplasma gondii obtained from aborted fetuses in an abortion outbreak in goats from northeastern Brazil. Brain samples from 32 fetuses were submitted to mouse bioassay for T. gondii isolation. Two isolates were obtained and subjected to genotypic characterization. Isolate virulence was evaluated using murine model in different doses (from 105 to 101 tachyzoites/mL). In genotyping, both isolates were classified as clonal lineage type II (genotype #1 ToxoDB) and showed to be virulent for mice. This is the first description of genotype #1 in cases of goat abortion, showing the circulation of virulent T. gondii isolate producing reproductive disorders in pregnant goat.

Caprine toxoplasmosis in Southern Brazil: a comparative seroepidemiological study between the indirect immunofluorescence assay, the enzyme-linked immunosorbent assay, and the modified agglutination test.

This study investigated the prevalence of caprine toxoplasmosis in goat herds from Southern Brazil by the indirect immunofluorescence assay (IFA), and compared these results with the enzyme-linked immunosorbent assay (ELISA) and the modified agglutination test (MAT). In addition, possible risk factors associated with infection due to Toxoplasma gondii were determined. The serum samples evaluated were from 1,058 goats derived from 94 goat herds within the state of Paraná, Southern Brazil. Seropositivity by IFA was 30.0%, 33.3% by ELISA, and 25.3% by MAT. The risk factors associated with infection by T. gondii in goats were pasture rental, female goats, the presence of cats, and pastures shared with several goat herds. Using IFA as a standard, ELISA and MAT showed substantial concordance (kappa = 0.74 and 0.61), with sensitivities of 87 and 66.2% and specificities of 89.7 and 92.2%, respectively. These results demonstrate that caprine toxoplasmosis is endemic within the state of Paraná, Southern Brazil. In addition, the results from the three assays were relevant, without any significant differences as demonstrated due to the substantial concordance based on the kappa index.

Co-Infection by Sarcoptes scabiei and Microsporum gypseum in Free-Ranging Crab-Eating Fox, Cerdocyon thous (Linnaeus, 1766)

ABSTRACT The aim of the present study was to describe the clinical manifestation, treatment and outcome of a case of co- infection by Sarcoptes scabiei and Microsporum gypseum in Cerdocyon thous (crab-eating fox) from Northeastern Brazil.

An outbreak of caprine toxoplasmosis - investigation and case report / Surto de toxoplasmose caprina - investigação e relato de caso

ABSTRACT: The present study aimed to investigate an abortion outbreak in a dairy goat herd in the municipality of Arapoti, Parana, Brazil. At the beginning of the outbreak, blood samples were collected from 33 goats with clinical signs; later, of the whole goat herd, two cats and two dogs. Milk samples were collected from 78 lactating goats. Four environmental soil samples and four samples of feed residue from goat feeders were collected too. Immunofluorescence antibody test (IFA) was used for serodiagnosis, the molecular analysis was conducted by means of the polymerase chain reaction (PCR), for the isolation of the etiological agent the bioassay was used. The results of the IFA revealed that 76.53% (137/179) of the goats, two dogs and two cats were seropositive for Toxoplasma gondii. Bioassay revealed one buffy coat and two milk sample having viable T. gondii. In the PCR, 11 whole blood samples, eight milk, three feeder troughs, and all soil samples were positive. The findings of the present study confirmed an outbreak caused by environmental contamination (of soil and feed) with T. gondii oocysts that could have been shed by kittens that lived on the farm and had access to the stock of goat food, facilitating this contamination, which reinforces the need for veterinary assistance and good management practices on farms.

RESUMO: O presente estudo teve como objetivo investigar um surto de aborto em um rebanho de cabras leiteiras no município de Arapoti, Paraná, Brasil. No início do surto, foram coletadas amostras de sangue de 33 cabras com sinais clínicos; mais tarde, de todo o rebanho caprino, dois gatos e dois cachorros. Foram obtidas amostras de leite das 78 cabras em lactação. Quatro amostras ambientais de solo e quatro de resíduos de comedouro também foram coletadas. O teste de imunofluorescência (IFI) foi utilizado para o sorodiagnóstico, a análise molecular foi conduzida por meio da reação em cadeia da polimerase (PCR), para isolamento do agente etiológico utilizou-se o bioensaio. Os resultados da IFI revelaram que 76,53% (137/179) das cabras, todos os cães e gatos eram soropositivos para Toxoplasma gondii. O bioensaio revelou uma amostra de camada leucocitária e duas de leite contaminadas com T. gondii viável. Na PCR, 11 amostras de sangue total, oito de leite, três resíduos alimentares e todas as amostras de solo foram positivas. Os resultados do presente estudo confirmaram um surto causado por contaminação ambiental (de solo e alimentos) com oocistos de T. gondii que, provavelmente, foram eliminados por gatos que permaneceram na fazenda e tinham acesso ao estoque de alimento dos caprinos, reforçando a necessidade de assistência técnica veterinária e boas práticas de manejo.

Toxoplasma gondii in backyard pigs: seroepidemiology and mouse bioassay.

The aim of this study was to evaluate the occurrence of Toxoplasma gondii in backyard pigs destined for human consumption in Pernambuco state, Brazil. Blood and tissue samples (liver, heart, brain, lung and diaphragm) were collected from 224 pigs from legal slaughterhouses and tested for T. gondii infection. Antibodies to T. gondii were found in the sera of 37.9% (85/224) by using the immunofluorescence antibody test (cut-off - 1:64). Tissue samples from seropositive pigs were bioassayed in mice. Tissue samples from seropositive pigs and from mice of the bioassay were submitted to histopathology, immunohistochemistry, polymerase chain reaction (PCR) and sequencing; 14.1% of pig tissue samples and 27.7% of bioassayed mouse samples were positive for T. gondii DNA, but all pig and mouse tissues were negative in histopathology analysis and immunochemistry. By using a risk assessment questionnaire, there was significant difference (p<0.001) in seroprevalence of 21.2% (reproducer) and 3.1% (finishing pig). These data serve as indicative of the sanitary conditions and risk of T. gondii infection for backyard pigs. Preventive measures must be implemented by health services to avoid toxoplasmosis human cases due to ingestion of pig meat.

Cryptosporidium spp. and Giardia spp. in feces and water and the associated exposure factors on dairy farms.

The aims of this study were to verify the prevalence of Cryptosporidium spp. and Giardia spp. in animal feces and drinking water on dairy farms and to identify a possible relation between the exposure factors and the presence of these parasites. Fecal samples from cattle and humans and water samples were collected on dairy farms in Paraná, Brazil. Analysis of (oo)cysts in the feces was performed by the modified Ziehl-Neelsen staining and centrifugal flotation in zinc sulfate. Test-positive samples were subjected to nested PCR amplification of the 18SSU ribosomal RNA gene for identification of Cryptosporidium and Giardia and of the gp60 gene for subtyping of Cryptosporidium. Microbiological analysis of water was carried out by the multiple-tube method and by means of a chromogenic substrate, and parasitological analysis was performed on 31 samples by direct immunofluorescence and nested PCR of the genes mentioned above. Identification of the species of Cryptosporidium was performed by sequencing and PCR with analysis of restriction fragment length polymorphisms. The prevalence of Giardia and Cryptosporidium was higher in calves than in adults. Among the samples of cattle feces, Cryptosporidium parvum was identified in 41 (64%), C. ryanae in eight (12.5%), C. bovis in four (6.3%), C. andersoni in five (7.8%), and a mixed infection in 20 samples (31.3%). These parasites were not identified in the samples of human feces. Thermotolerant coliform bacteria were identified in 25 samples of water (45.5%). Giardia duodenalis and C. parvum were identified in three water samples. The gp60 gene analysis of C. parvum isolates revealed the presence of two strains (IIaA20G1R1 and IIaA17G2R2) in the fecal samples and one (IIaA17G2R1) in the water samples. The presence of coliforms was associated with the water source, structure and degradation of springs, rain, and turbidity. The prevalence of protozoa was higher in calves up to six months of age. C. parvum and G. duodenalis were identified in the water of dairy farms, as were thermotolerant coliforms; these findings point to the need for guidance on handling of animals, preservation of water sources, and water treatment.

rROP2 from Toxoplasma gondii as a potential vaccine against oocyst shedding in domestic cats.

The aim of the present study was to evaluate oocyst shedding in cats immunized by nasal route with T. gondii proteins ROP2. Twelve short hair cats (Felis catus) were divided in three groups G1, G2 and G3 (n=4). Animals from G1 received 100 µg of rROP2 proteins plus 20 µg of Quil-A, G2 received 100 µg of BSA plus 20 µg of Quil-A, and the G3 only saline solution (control group). All treatments were done by intranasal route at days 0, 21, 42, and 63. The challenge was performed in all groups on day 70 with ≅ 800 tissue cysts of ME-49 strain by oral route. Animals from G1 shed less oocysts (86.7%) than control groups. ELISA was used to detect anti-rROP2 IgG and IgA, however, there were no correlation between number of oocyst shedding by either IgG or IgA antibody levels. In the present work, in spite of lesser oocysts production in immunized group than control groups, it was not possible to associate the use of rROP2 via nostrils with protection against oocyst shedding. For the future, the use of either other recombinant proteins or DNA vaccine, in combination with rROP2 could be tested to try improving the efficacy of this kind of vaccine.

rROP2 from Toxoplasma gondii as a potential vaccine against oocyst shedding in domestic cats / rROP2 de Toxoplasma gondii como potencial vacina contra a eliminação de oocistos em gatos domésticos

Abstract The aim of the present study was to evaluate oocyst shedding in cats immunized by nasal route with T. gondii proteins ROP2. Twelve short hair cats (Felis catus) were divided in three groups G1, G2 and G3 (n=4). Animals from G1 received 100 μg of rROP2 proteins plus 20 μg of Quil-A, G2 received 100 μg of BSA plus 20 μg of Quil-A, and the G3 only saline solution (control group). All treatments were done by intranasal route at days 0, 21, 42, and 63. The challenge was performed in all groups on day 70 with ≅ 800 tissue cysts of ME-49 strain by oral route. Animals from G1 shed less oocysts (86.7%) than control groups. ELISA was used to detect anti-rROP2 IgG and IgA, however, there were no correlation between number of oocyst shedding by either IgG or IgA antibody levels. In the present work, in spite of lesser oocysts production in immunized group than control groups, it was not possible to associate the use of rROP2 via nostrils with protection against oocyst shedding. For the future, the use of either other recombinant proteins or DNA vaccine, in combination with rROP2 could be tested to try improving the efficacy of this kind of vaccine.

Resumo O objetivo do presente estudo foi avaliar a eliminação de oocistos de Toxoplasma gondii em gatos imunizados pela via nasal com proteínas ROP2 de T. gondii. Doze gatos sem raça definida (Felis catus) foram divididos em três grupos experimentais G1, G2 e G3 (n = 4). Os animais do G1 receberam 100 μg de proteínas de rROP2 mais 20 μg de Quil-A, G2 recebeu 100 μg de albumina de soro bovino (BSA) junto com 20 μg de Quil-A, e o G3 recebeu apenas solução salina (grupo de controle). Todos os tratamentos foram realizados pela via intranasal nos dias 0, 21, 42 e 63. O desafio foi realizado em todos os grupos no dia 70 com aproximadamente 800 cistos de tecido da cepa ME-49 por via oral. Os animais de todos os grupos tiveram as suas fezes examinadas e o número de oocistos foi determinado durante 20 dias após o desafio. Os animais de G1 eliminaram menos oocistos (86,7%) do que os grupos controles. O ELISA foi utilizado para detectar IgG e IgA anti-rROP2, no entanto, não houve correlação entre o número de eliminhação de oocistos com os níveis de anticorpos IgG ou IgA. No presente trabalho, apesar da menor produção de oocistos no grupo imunizado (G1) em relação aos grupos controles (G2 e G3), não foi possível associar o uso de rROP2 pela via nasal com proteção contra eliminação de oocistos de T. gondii. Para o futuro, a utilização de outras proteínas recombinantes, ou mesmo vacina de DNA, em combinação com rROP2 poderia ser utilizada para tentar melhorar a eficácia deste tipo de vacina.

First isolation and RFLP genotyping of Toxoplasma gondii from crab-eating fox (Cerdocyon thous-Linnaeus, 1766).

Wild animals may play an important role in the transmission and maintenance of Toxoplasma gondii in the environment. The purpose of the present study was to isolate and genotype T. gondii from a free-ranging crab-eating fox (Cerdocyon thous-Linnaeus, 1766). A crab-eating fox in critical health condition was attended in a veterinary hospital in Recife, Pernambuco State, Brazil. The animal died despite emergency treatment. The brain was collected aseptically and destined for mouse bioassay. One isolate of T. gondii was obtained, and Polymerase Chain Reaction - Restriction Fragment Length Polymorphism (PCR-RFLP) was used to assess genetic variability at 11 markers (SAG1, SAG2, altSAG2, SAG3, BTUB, GRA6, c228, c292, L358, PK1 and APICO). A murine model was used to assess the virulence of the isolate. Using the PCR-RFLP, genotype ToxoDB #13 was identified, which is considered an atypical strain. The isolate was classified as avirulent in the murine model. This is the first study to report T. gondii infection in the crab-eating fox.

Comparison of indirect fluorescent antibody test and the modified agglutination test for the detection of Toxoplasma gondii antibodies in stray dogs from Southern Brazil.

The aim of the present study was to determine the prevalence of antibodies to Toxoplasma gondii by two serological techniques in sera of 364 stray dogs from Brazil by immunofluorescence antibody test (IFAT, cut off point 1:16) and to the modified agglutination test (MAT, cut-off points 1:25 and 1:50). A total of 175/364 (48.07%) sera were positive by IFAT, and 108/364 (29.67%) and 85/364 (23.35%) were positive by MAT with cutoff points 1:25 and 1:50, respectively were positive by MAT. Cohen's Kappa Coefficient between IFAT and MAT was 0.81 (excellent) and 0.66 (substantial) with cutoff points 1:25 and 1:50, respectively. Using IFAT as gold standard, MAT sensitivity and specificity were 78% and 99% for 1:25 and 61% and 99% for 1:50, respectively. The results document of the usefulness of MAT for serological diagnosis because it does not require species-specific conjugate.

Application of different techniques to detect Toxoplasma gondii in slaughtered sheep for human consumption.

The aim of this study was to investigate occurrence of Toxoplasma gondii in sheep slaughtered in the state of Alagoas, Brazil, by means of different diagnosis techniques. Serum samples and tissues from 100 slaughtered sheep were used. To detect antibodies, the indirect immunofluorescence antibody test (IFAT) was used, and tissues from seropositive animals (cut-off ≥1:64) were submitted to Polymerase Chain Reaction (PCR) and immunohistochemistry (IHC). To assess the concordance between the direct techniques, the kappa test was used. In the IFAT, it was observed that 14% (14/100) of the ovine samples were serum-positive. In the PCR, 21.43% (3/14) of the animals were positive and in IHC, it was observed that 7.14% (1/14) were positively stained for T. gondii in cerebral tissue. Histopathologically, the predominant finding was the presence of mononuclear cell infiltrate in the heart and a perivascular cuff in the cerebrum and cerebellum. The concordance between the direct diagnosis techniques was moderate (k=0.44). Thus, it is important to use different direct techniques in diagnosing toxoplasmosis in naturally infected sheep.

Application of different techniques to detect Toxoplasma gondii in slaughtered sheep for human consumption / Utilização de diferentes técnicas para detecção de Toxoplasma gondii em ovinos abatidos para consumo humano

Abstract The aim of this study was to investigate occurrence of Toxoplasma gondii in sheep slaughtered in the state of Alagoas, Brazil, by means of different diagnosis techniques. Serum samples and tissues from 100 slaughtered sheep were used. To detect antibodies, the indirect immunofluorescence antibody test (IFAT) was used, and tissues from seropositive animals (cut-off ≥1:64) were submitted to Polymerase Chain Reaction (PCR) and immunohistochemistry (IHC). To assess the concordance between the direct techniques, the kappa test was used. In the IFAT, it was observed that 14% (14/100) of the ovine samples were serum-positive. In the PCR, 21.43% (3/14) of the animals were positive and in IHC, it was observed that 7.14% (1/14) were positively stained for T. gondii in cerebral tissue. Histopathologically, the predominant finding was the presence of mononuclear cell infiltrate in the heart and a perivascular cuff in the cerebrum and cerebellum. The concordance between the direct diagnosis techniques was moderate (k=0.44). Thus, it is important to use different direct techniques in diagnosing toxoplasmosis in naturally infected sheep.

Resumo O objetivo deste estudo foi pesquisar a ocorrência de Toxoplasma gondii em ovinos abatidos no Estado de Alagoas, Brasil por meio de diferentes técnicas de diagnóstico. Foram utilizadas amostras de soros e tecidos de 100 ovinos abatidos. Para a pesquisa de anticorpos foi utilizada a Reação de Imunofluorescência Indireta (RIFI), e os tecidos dos animais soropositivos (ponto de corte ≥1:64) foram submetidos às técnicas de Reação de Cadeia da Polimerase (PCR) e Imunohistoquímica (IHQ). Para o estudo da concordância entre as técnicas diretas foi empregado o teste Kappa. Na RIFI, 14% (14/100) das amostras foram soro-positivas. Na PCR, 21,43% (3/14) dos animais foram positivos e, na IHC, 7,14% (1/14) apresentaram marcação positiva para T. gondii no tecido cerebral. Na histopatologia, o achado predominante foi o infiltrado celular mononuclear no coração e manguito perivascular no cérebro e cerebelo. A concordância entre as técnicas diretas de diagnóstico foi moderada (K= 0,44). Desse modo, é importante utilizar diferentes técnicas diretas no diagnóstico da toxoplasmose em ovinos naturalmente infectados.

Occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system, Paraná, Southern Brazil / Ocorrência de Cryptosporidium spp. e Giardia spp. em uma estação pública de tratamento de água, Paraná, Sul do Brasil

The purpose of this study was to investigate the occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system. Samples of raw and treated water were collected and concentrated using the membrane filtration technique. Direct Immunofluorescence Test was performed on the samples. DNA extraction using a commercial kit was performed and the DNA extracted was submitted to a nested-PCR reaction (n-PCR) and sequencing. In the immunofluorescence, 2/24 (8.33%) samples of raw water were positive for Giardia spp.. In n-PCR and sequencing, 2/24 (8.33%) samples of raw water were positive for Giardia spp., and 2/24 (8.33%) samples were positive for Cryptosporidium spp.. The sequencing showed Cryptosporidium parvum and Giardia duodenalis DNA. In raw water, there was moderate correlation among turbidity, color and Cryptosporidium spp. and between turbidity and Giardia spp.. The presence of these protozoans in the water indicates the need for monitoring for water-treatment companies.

O objetivo deste estudo foi investigar a ocorrência de Cryptosporidium spp. e Giardia spp. em um sistema público de tratamento de água. Amostras de água bruta e tratada foram coletadas e concentradas, utilizando-se a técnica de filtração em membranas. Foi realizada a técnica de Imunofluorescência Direta nas amostras. A extração de DNA foi realizada, utilizando-se um kit comercial, e o DNA extraído foi submetido a uma reação de nested-PCR (n-PCR). Na imunofluorescência, 2/24 (8,33%) amostras de água bruta foram positivas para Giardiaspp.. Na n-PCR, 2/24 (8,33%) amostras de água bruta foram positivas para Giardia spp., e 2/24 (8,33%) amostras foram positivas para Cryptosporidium spp.. O sequenciamento demonstrou DNA de Cryptosporidium parvum e de Giardia duodenalis. Na água bruta, houve correlação moderada entre turbidez, cor e Cryptosporidium spp. e entre a turbidez e Giardia spp.. A presença desses protozoários na água indica a necessidade de monitoramento pelas empresas de tratamento de água.

First report of genotype #65 of Toxoplasma gondii in pigs.

The aim of the present study was to isolate and genotype Toxoplasma gondii from pigs slaughtered for human consumption in northeastern Brazil. Indirect immunofluorescence antibody test (IFAT) was used to screen positive pigs. Tissues samples of animals with antibody titers ≥64 were submitted to bioassay in mice. One isolate of T. gondii was obtained, and the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique, using 11 markers (SAG1, SAG2, altSAG2, SAG3, BTUB, GRA6, c228, c292, L358, PK1, and APICO), was applied to evaluate the genetic variability. DNA from reference strains was used as a positive control. By means of genetic analysis, genotype ToxoDB #65 was identified, which is considered an atypical strain. This is the first record of genotype #65 in pigs. Thus, further studies in this region are necessary to determine the genetic variability of T. gondii in pigs and possible impact on public health.

Occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system, Paraná, Southern Brazil.

The purpose of this study was to investigate the occurrence of Cryptosporidium spp. and Giardia spp. in a public water-treatment system. Samples of raw and treated water were collected and concentrated using the membrane filtration technique. Direct Immunofluorescence Test was performed on the samples. DNA extraction using a commercial kit was performed and the DNA extracted was submitted to a nested-PCR reaction (n-PCR) and sequencing. In the immunofluorescence, 2/24 (8.33%) samples of raw water were positive for Giardia spp.. In n-PCR and sequencing, 2/24 (8.33%) samples of raw water were positive for Giardia spp., and 2/24 (8.33%) samples were positive for Cryptosporidium spp.. The sequencing showed Cryptosporidium parvum and Giardia duodenalis DNA. In raw water, there was moderate correlation among turbidity, color and Cryptosporidium spp. and between turbidity and Giardia spp.. The presence of these protozoans in the water indicates the need for monitoring for water-treatment companies.

Ocorrência de Cryptosporidium spp. em dois centros de treinamento de equinos de Curitiba, Paraná / Occurrence of Cryptosporidium spp. in two centers of training horses in Curitiba, Paraná

O presente trabalho investigou a ocorrência de parasitismo por Cryptoporidium spp. em equinos alojados em dois centros de treinamento de equinos localizados no municipio de Curitiba, Paraná. Foram examinados 108 cavalos, sendo 48 procedentes do Centro de treinamento 1 (CT1) e 60 do Centro de Treinamento 2 (CT2). As coletas de amostras de fezes foram realizadas no período de outubro de 2010 a janeiro de 2011. A metodologia utilizada para a confirmação da presença de oocistos de Cryptosporidium spp. foi a técnica de Ziehl-Neelsen modificada. A ocorrência encontrada foi de 18,52% para o total de animais examinados. Houve diferença estatística significativa (p > 0,05) quando comparadas as prevalências encontradas nos dois centros de treinamento, sendo no CT1 de 4,16% e no CT2 de 30%. Não houve associação entre a prevalência e a idade, o sexo e raça (p > 0,05).

This study investigated the occurrence of parasitism by Cryptosporidium spp. in horses housed in two training centers located in Curitiba, Paraná. A total of 108 horses were examined, 48 from the Training Center 1 (CT1) and 60 of the Training Centre 2 (CT2). Collection of stool specimens occurred from October 2010 to January 2011. The methodology used to confirm the presence of Cryptosporidium spp. was the Ziehl-Neelsen modified. The occurrence found was 18.52% for the total animals examined. There was a statistically significant difference (p > 0.05) comparing prevalence rates in the two training centers: 4.16% in CT1 and 30% in CT2. There was no association between prevalence and the age, sex and race (p > 0.05) of the horses.

Perfil epidemiológico de casos de surtos de doenças transmitidas poralimentos ocorridos no Paraná, Brasil / Epidemiological profile of foodborne diseases outbreaks occurred in Parana state, Brazil

A investigação dos surtos de Doenças Transmitidas por Alimentos (DTA) é desafiadora. Para a Vigilância em Saúde é fundamental a identificação de grupos e fatores de risco associados às DTA. O objetivo deste trabalho foi caracterizar o perfil epidemiológico dos surtos de DTA ocorridos nos 29 municípiosque compõe a 2ª Regional de Saúde, da Secretaria Estadual de Saúde do Paraná, Brasil, entre 2005-2008. Tabularam-se 472 laudos de alimentos do Laboratório Central do Estado do Paraná (LACEN-PR) e fichas notificadas no Sistema de Informação de Agravos de Notificação (SINAN) de casos suspeitos de DTA. O grupo mais acometido foi de indivíduos do sexo feminino com 50,51% dos casos. Indivíduos entre 20 a 49 anos de idade foram os mais frequentemente envolvidos com 49,50% das ocorrências. Restaurantes (27,65%) e residências dos indivíduos acometidos (36,17%) foram os locais de preparo econsumo mais frequentemente envolvidos. Manipulação/preparo inadequados dos alimentos (36,95%) eforma de consumo inadequada (23,90%) foram os fatores causais mais predominantes. A maior parte dasconfirmações dos surtos fundamentou-se em achados clínico-epidemiológicos (44,68%). Com relaçãoaos micro-organismos identificados nas amostras analisadas, os resultados apontaram predominância Escherichia coli (indicador sanitário), seguido pelo Bacillus cereus e Staphyloccus aureus. Os alimentos mais frequentemente envolvidos foram o leite (29,68%) e as frutas, verduras e legumes (24,21%). Com a pesquisa é possível orientar medidas preventivas e de controle de DTA na população pesquisada.

The investigation of outbreaks of Food Borne Diseases (FBD) is challenging. For the Health Surveillance is essential to identify groups and risk factors associated with the DTA. The aim of this study was to characterize the epidemiology of outbreaks of DTA occurred in 29 counties that comprise the 2nd Regional Health, the Department of Health of Paraná, Brazil, from 2005-2008. Tabulated to 472 reports of food the Central Laboratory of the State of Paraná (LACEN-PR) chips and notified the Information System for Notifiable Diseases (SINAN) of suspected cases of DTA. The group most affected were females with 50.51% of the cases. Individuals between 20 and 49 years of age were most often involved in 49.50% of cases. Restaurants (27.65%) of affected individuals and households (36.17%) were the preparation and consumption sites most frequently involved. Manipulation / inadequate preparation of food (36.95%) and inadequate form of consumption (23.90%) were the most prevalent causative factors.Most confirmations of outbreaks was based on clinical and epidemiological (44.68%). With respect to micro-organisms identified in the samples analyzed, the results showed predominance Escherichia coli(health indicator), followed by Bacillus cereus and Staphylococcus aureus. The foods most frequently involved were milk (29.68%) and fruits and vegetables (24.21%). With this study can guide preventive measures and control of DTA in the population studied.