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In this study, we aimed to investigate an outbreak of abortion in a herd of beef sheep on a property in Ivaiporã, north central Paraná, Brazil. A total of 64 blood samples were collected from the sheep, 33 of which were mothers with reports of abortion or stillbirth, and 7 with weak lamb birth. For epidemiological investigation of the parasite, fourteen samples were collected from different sources on the property, six from water, three from soil, one from trough, and four from feed. Indirect immunofluorescence (IFI) analysis and polymerase chain reaction were performed for detecting anti-Toxoplasma gondii antibodies in the sheep and molecular identification of the protozoan in the environment, respectively. The IFI results showed that 95.3% of the sheep were seropositive for anti-T. gondii antibodies; however, the environmental samples tested negative. During the visit to the property and interview with the owner, free-roaming cats were observed on the property for biological control of rodents that had access to the food deposits. This type of management may have triggered the outbreak, which reinforces the importance of preventive management and veterinary medical monitoring on sheep farms.

The present study aimed to molecularly characterize Giardia duodenalis from stool samples of humans, dogs, and cats. Molecular analyses were performed on 59 samples that tested positive for G. duodenalis on coproparasitological examinations. After extraction, the samples were first tested by nested polymerase chain reaction (n-PCR) analysis of the SSU-rRNA gene, and for the samples that were positive, the β-giardin, TPI, and GDH genes were analyzed. The amplicons obtained in the n-PCR of the β-giardin gene were subjected to PCR-restriction length polymorphism (RFLP) analysis and subsequent digestion with the enzyme HaeIII to differentiate the assemblages. Seven (11.8 %), 34 (57.7 %), and 18 (30.5 %) out of 59 samples were from humans, dogs, and cats, respectively. Nested-PCR results showed that 49.2 % (29/59) of samples were positive for the SSU-rRNA gene, with 42.9 % (3/7) of humans, 55.9 % (19/34) of dogs, and 38.9 % (7/18) of catsve. Of the other genes analyzed, β-giardin was amplified most frequently, in 34.5 % (10/29) of samples, followed by GDH in 27.6 % (8/29) of samples, and TPI in 10.3 % (3/29) of samples. Only one sample from a dog showed the amplification of all genes. PCR-RFLP analysis showed assemblage F in a human, dog, and cat samples; and assemblage C and D in dog samples. This is the first description of assemblage F in humans from Brazil and the first description of assemblage F in dogs. Further studies are needed to verify the frequency with which these infections occur, and provide information that will contribute to the molecular epidemiological understanding of giardiasis.

Pyothorax is the accumulation of exudade in the pleural cavity. Affected patients are often presented with acute dyspnea. We report a case of pyothorax associated with the presence of a bone fragment in the lower respiratory tract of an adult domestic cat. We presume that the bone fragment reached the respiratory tract by either aspiration or migration from the abdominal cavity. The patient presented abdominal distension and acute dyspnea. Bloodwork revealed neutrophilic leukocytosis with left shift and lymphocytosis. Thoracic fluid analysis confirmed septic exudate. The patient was unresponsive to the treatment and died within 24 hours of admission. Necropsy revealed the presence of a bone fragment in the lower respiratory tract. We suggest that foreign bodies should be considered as a rule-out cause of pyothorax and sepsis in domestic cats.

Cryptosporidium e Giardia são protozoários que causam gastroenterite em seus hospedeiros, e as espécies mais frequentes no mundo são as de potencial zoonótico. A água é majoritariamente contaminada por esgoto doméstico e escoamentos agropecuários, sendo a via de transmissão de maior preocupação à saúde da população. Dentre as fontes de contaminação para a água, ambos os protozoários são frequentemente encontrados no esgoto, todavia, poucos estudos relacionam dados sobre viabilidade. Para detecção de viabilidade, o método de coloração com Propídio-monoazida (PMA) associada a PCR é avaliada como alternativa a técnica de imunofluorescência associada ao corante 4?,6-diamidino-2-phenylindole (DAPI). O conhecimento da viabilidade desses agentes em esgoto é necessário para avaliar os riscos de contaminação da água e consequentemente, a situação de saúde da população. Os objetivos do estudo foram verificar a viabilidade de Cryptosporidium spp. e Giardia spp. e quantificar oocistos viáveis em afluente e efluente de Estação de Tratamento de Esgoto (ETE) no interior do Paraná. De setembro a novembro de 2020 foram realizadas 10 coletas do afluente e efluente da ETE com periodicidade semanal, desse modo foi possível acompanhar sete processos de tratamento desde a da chegada até a saída do esgoto, conforme o tempo de detenção hidráulica. As amostras do afluente e efluente foram concentradas por centrifugação associada a filtração e purificadas por flutuação em solução saturada de sacarose, para aprimorar a sensibilidade de detecção. A viabilidade foi verificada por marcação do DNA das células inviáveis com corante PMA, após fotoativação, associada a detecção de Cryptosporidium e Giardia por nPCR e quantificação de oocistos por qPCR. Todas as amostras de afluente da ETE foram positivas para os protozoários com a presença de formas viáveis, com média de 8.769 oocistos de Cryptosporidium viáveis/L. Em efluente, todas foram positivas para Cryptosporidium com concentração média de 1.855 oocistos viáveis/L e para Giardia, 70% foram positivas, com formas viáveis em 30% delas. O acompanhamento do tratamento, de acordo com tempo de detenção, demonstrou persistência média de 31,17% dos oocistos viáveis após tratamento. Os resultados apresentam uma provável situação endêmica dos protozoários na população que abastece a ETE com alta concentração de oocistos de Cryptosporidium spp. O tratamento do esgoto reduziu a frequência de cistos de Giardia e concentração de oocistos de Cryptosporidium, porém não foi capaz de inviabilizar todos os (oo)cistos. Dessa forma, o corpo hídrico recebe cargas frequentes de formas viáveis desses protozoários, que podem causar prejuízo a saúde. A variação na eficácia da redução de oocistos viáveis, demonstrada ao longo dos acompanhamentos de tratamento, evidencia a necessidade de monitoramento da viabilidade desses patógenos no efluente de esgoto para predizer e prevenir riscos de infecção da população. Cryptosporidium and Giardia are protozoa which cause gastroenteritis in their hosts. The zoonotic species are the most frequent in the world. The major sources of contamination to water are runoffs from livestock and domestic wastewater and water is the source of transmission of most worry for health risks. The protozoa are frequently found in wastewater, although, few studies include viability data of (oo)cysts. The Propidium monoazide (PMA) stain method associated to PCR is assessed as an alternative to immunofluorescence and 4?,6-diamidino-2-phenylindole (DAPI) stain for viability detection of (oo)cysts. The knowledge of viability in sewage is necessary to assess the status of population health and water risk of contamination. The aims of the study were to verify the viability of Cryptosporidium and Giardia and quantify viable oocysts in influent and effluent of a Wastewater Treatment Plant (WWTP) in interior of Paraná state of Brazil. Ten collects were executed from September to November 2020 weekly. Respecting the hydraulic detention time, seven monitoring of the wastewater treatment since the sewage arrived until its left of the station were possible. The samples concentration was done by centrifugation associated with filtration and purification for improvement in sensibility. The viability was verified by PMA stain in inviable cells and photoactivation. It was associated to nPCR for the detection of Cryptosporidium and Giardia and to qPCR for the quantification of viable oocysts. All the WWTP influent samples were positive for both protozoa including the presence of viable oocyst. The mean concentration of Cryptosporidium oocysts was 8,769 oocysts/L. In the WWTP effluent, all the samples were positive to Cryptosporidium with a mean concentration of 1,855 viable oocysts/L and 70% of samples were positive for Giardia in which 30% had viable cysts. The monitoring of wastewater treatment demonstrated a mean persistence of 31,17% of viable oocysts after treatment. The results indicated a probable endemic presence of these protozoa in the population which supplies the WWTP and Cryptosporidium presents high concentration of viable forms. The wastewater treatment reduced the frequency of cysts and the concentration of oocysts, although it was not capable to make all (oo)cysts unviable. The receptor river receives frequent charges of viable (oo)cysts which may cause risks for health. The variation in the reduction efficacy of viable oocysts evidences the necessity of monitoring (oo)cysts viability in sewage effluent to predict and prevent infection risks to the population.

This study describes the etiological diversity observed in a severe neonatal diarrhea outbreak with morbidity and mortality rates of 80 and 20%, respectively, with detection of mixed infections with viral, bacterial, and protozoan disease agents in a dairy calf rearing unit. Diarrheic fecal samples were collected from eight 5 to 18 days of age calves and were submitted to the investigation of the presence of rotavirus A (RVA), bovine coronavirus (BCoV), bovine kobuvirus (BKV), bovine viral diarrhea virus 1 and 2 (BVDV-1 and BVDV-2), enteropathogenic Escherichia coli (ETEC), Salmonella sp., and Cryptosporidium spp. Fragments of the small intestine of one calf with diarrhea that spontaneously died were submitted for histopathological analyses. The most frequent infectious agent detected in diarrheic fecal samples was BKV (8/8—100%), followed by RVA (5/8—62.5%), BVDV (5/8—62.5%), Cryptosporidium parvum (5/8—62.5%), ETEC (4/8—50%), and Cryptosporidium ryanae (1/8—12.5%). These etiological agents were found in mixed infections with two or more pathogens per diarrheic fecal sample. The association of viral and protozoan pathogens was the most frequently identified (37.5%) in these samples, followed by viral and bacterial (25%); viral, bacterial, and protozoan (25%); and only viral agents (12.5%). BCoV and Salmonella sp. were not identified in the diarrheic fecal samples analyzed. Additionally, histopathology of the small intestine diagnosed chronic lymphocytic enteritis. In conclusion, in calf rearing units, the adoption and strict monitoring of health management practices are critical to the success of this calf creation system. Supplementary Information The online version contains supplementary material available at 10.1007/s42770-021-00565-5.

Cryptosporidium and Giardia are protozoan parasites that cause diarrhea in humans and animals. Molecular characterization of these pathogens in sewage may provide insight on their occurrence and prevalence in Brazil. This study aimed to investigate the presence of Giardia and Cryptosporidium in raw and treated sewage from Londrina, Paraná, Brazil. Samples were collected every two weeks during a year. Samples were concentrated, then DNA was extracted and subjected to a nested PCR targeting the Giardia 18S rRNA gene and the Cryptosporidium 18S rRNA gene. Species of Cryptosporidium were characterized by restriction fragment length polymorphism (RFLP). All raw sewage and 76% of the treated sewage were positive for Giardia; 84% of raw sewage samples and 8% of treated sewage were positive for Cryptosporidium. C. muris, C. hominis, C. baileyi, C. parvum and C. suis were detected in 100%, 19%, 9%, 9% and 4% of raw sewage, respectively. C. muris was the only species found in treated sewage. Multiple species of Cryptosporidium were present in 19.04% of the raw sewage. Treated sewage water can pose a threat to human health. The speciation of Cryptosporidium revealed the presence of non-common zoonotic species as C. suis and C. muris. Resumo Cryptosporidium e Giardia são protozoários causadores de diarreia em animais e humanos. A caracterização molecular destes protozoários em esgoto pode prover dados ainda desconhecidos da ocorrência de espécies. O objetivo do presente estudo foi monitorar a ocorrência de Giardia e espécies de Cryptosporidium em esgoto bruto e tratado em uma estação de tratamento de esgoto (ETE) de Londrina, Paraná. Amostras de esgoto bruto e tratado foram coletadas no período de um ano, com periodicidade quinzenal. A ocorrência destes protozoários foi caracterizada por meio de concentração das amostras e posterior extração de DNA seguida de nested-PCR para amplificação de fragmentos dos genes 18S rRNA de Giardia e 18S rRNA de Cryptosporidium. A caracterização das espécies de Cryptosporidium foi realizada por meio de análise por polimorfismo de comprimento do fragmento de restrição (RFLP) dos produtos obtidos. Foram coletadas no total 25 amostras de cada, esgoto bruto e esgoto tratado. Para Giardia, todas as amostras de esgoto bruto e 76% das de esgoto tratado foram positivas. Cryptosporidium esteve presente em 84% das amostras de esgoto bruto e em 8% do tratado. No esgoto tratado foi encontrado apenas C. muris, já nas amostras de esgoto bruto foram encontradas cinco espécies: C. muris, C. hominis, C. baileyi, C. suis e C. parvum em 100%, 19%, 9%, 9% e 4%, respectivamente. A presença de espécies mistas foi observada em 19,04% das amostras. A presença de Giardia e Cryptosporidium em esgoto tratado pode pôr em risco a saúde humana. A discriminação de espécies de Cryptosporidium revelou a presença de espécies zoonóticas incomuns como C. suis e C. muris.

The purpose of this study was to monitor the presence of Cryptosporidium spp. and Giardia duodenalis in a water treatment plant (WTP) using settling sludge and backwash water (BW) samples in previous and post the deflocculation of polyaluminium chloride (PAC) flacks. Eleven collections were performed. BW and settling sludge (SSF) were concentrated by calcium carbonate flocculation, and another aliquot of settling sludge (SSC) by centrifugation. The samples were divided as follows: Group A, containing 33 samples without degradation of PAC flakes, and Group B, with degradation by alkalinization with 10 M NaOH. Sample DNA was extracted with a commercial kit, and nested polymerase chain reaction (PCR) was used to detect Cryptosporidium and G. duodenalis. All samples from Group A were negative for Cryptosporidium spp., and 6.1% (2/33) were positive for G. duodenalis in SSC samples. While the absence of Cryptosporidium may be due to a low contamination level of the water resource, the presence of G. duodenalis indicates contamination of the raw water. The detection of G. duodenalis in SSC samples indicates that this detection method was the most effective. The 33 samples from Group B were negative for both protozoa, probably due to the presence of aluminium and humic substances.