Search

Cryptosporidium spp. is one of the most important pathogens infecting nursing calves worldwide. This study aimed to investigate the intestinal microbiota of dairy calves during the first month of life and the impact of diarrhea caused by Cryptosporidium on a Brazilian farm. Fecal samples from 30 calves were collected during the first month of life, and fecal scores were recorded. Samples from the second, third, and fourth days of life were analyzed by DNA sequencing of the 16S rRNA gene. In addition, samples of sixteen calves positive for Cryptosporidium spp. were retrospectively chosen according to the development of diarrhea: four and two days before diarrhea, at the onset of diarrhea, after four days of diarrhea, at the end of diarrhea, and after six days of diarrhea resolution. Diarrhea was observed in all calves (100%), starting at day 5 of life, and all calves tested positive for Cryptosporidium in at least one sample. The microbiota richness increased with age but was retarded by diarrhea. Compositional changes associated with Cryptosporidium infection included increases in Fusobacterium, Prevotella , and Peptostreptococcus , as well as decreases in Collinsella and Lachnospiraceae. In conclusion, Cryptosporidium infection has the potential to decrease richness and change the composition of the intestinal microbiota of dairy calves.

The state of Paraná is home to three out of the five medically significant snake genera in Brazil and lacks of snakebite epidemiology studies. This study aimed to ascertain the spatial, environmental, and socioeconomic factors associated with snakebite risk by analyzing notification data of cases in the state of Paraná. Notification and socioeconomic data were gathered from the online platforms of the National System of Notifiable Diseases (SINAN) and the Brazilian Institute of Geography and Statistics (IBGE). Land cover and land use maps were obtained from the Mapbiomas platform in raster format and subsequently converted into vectors using QGis software. The proportions of land use and land cover in square kilometers (km 2 ) were then calculated. All acquired data were tabulated using Microsoft Excel 365 software. For spatial analysis, GeoDa software version 1.20 was utilized to calculate the Global and Local Moran indices, assessing spatial correlations. Between 2007 and 2021, 12,877 notifications were recorded, with an average incidence of 8.22/100,000 inhabitants in the state, 8166 (63.41%) caused by Bothrops, 1534 (11.91%) caused by Crotalus, 56 (0.43%) caused by Micrurus. 1703 (13.22%) caused by non-venomous snake species, and the remaining cases did not have the identified causative species. The incidents caused by Bothrops and Crotalus showed different distribution patterns. Spatial analysis revealed that key factors contributing to snakebite risk included the presence of native forests, mangroves, apicuns, and monospecific planted forests. The population group at the highest risk comprised rural residents and workers. Furthermore, the absence of basic sanitation and proper garbage collection and disposal exhibited positive correlations with snakebites. Conversely, intensive farming practices with substantial mechanization and pastures demonstrated negative spatial correlations. This study has enabled the identification of the primary factors associated with snakebite risk, facilitating more targeted efforts to prevent snakebite accidents among vulnerable populations., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Isabelli Sayuri Kono reports financial support was provided by Coordination of Higher Education Personnel Improvement., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Trichinella spp. are zoonotic parasites that are widely distributed in warm-blooded carnivores and omnivores, including humans. Until the present moment, Brazil has been considered by World Animal Health Organization free from the domestic cycle of trichinellosis, whereas the parasite's sylvatic cycle has the status of infection in limited zones. However, neighboring countries such as Argentina have reports of parasite larvae in the wild fauna. The present study aimed to determine the occurrence of Trichinella spp. in road-killed wild animals in Paraná, Brazil. Biological samples from 71 wild animals-29 Didelphis albiventris, 11 Nasua nasua, ten Cerdocyon thous, seven Dasypus novemcinctus, six Leopardus guttulus, six Sphiggurus spinosus and two Puma concolor-collected from November 2016 to November 2021 were subjected to artificial digestion, following the methodology described in the REGULATION (EC) No. 2075/2005. No Trichinella spp. larvae were detected in the carcasses of the road-killed wild animals. However, considering the wide spectrum of possible reservoirs that could act as a link between the sylvatic and domestic cycles and considering the current Brazilian status of sylvatic trichinellosis in limited zones, frequent monitoring of wild fauna remains necessary.

Cryptosporidium and Giardia are the main etiologies of waterborne outbreaks caused by protozoa. These parasites are commonly detected in wastewater; however, there is little knowledge about the concentration of viable forms in treated sewage, mainly in small communities. To understand more about the presence of viable oocysts and cysts in domestic sewage, we monitored the affluent and effluent of a wastewater treatment plant (WWTP) in inner-city Brazil. Ten samplings and seven follow-ups were performed in 2020. Samples were concentrated by centrifugation, filtration and purified by fluctuation. Viability was accessed by propidium-monoazide (PMA) associated with nPCR and qPCR. Both viable protozoa were detected in all raw sewage samples (average: 438.5 viable oocysts/L). Regarding treated sewage, Cryptosporidium was detected in all of the samples (average: 92.8 viable oocysts/L) and Giardia was detected in 70% with viable cysts in 30%. Considering the follow-ups, 31.17% of Cryptosporidium viable oocysts remained in the effluent after the treatment. High amounts of Cryptosporidium and a high frequency of Giardia were detected, therefore both arrived at WWTP and were discharged into the river. These alert the presence of agro-industrial effluents into domestic sewage and demonstrated the effectiveness of the concentration technique for monitoring protozoa in wastewater.

The intense contact of children with domestic animals or environments contaminated with faeces of these animals, together with habits related to lack of hygiene, can facilitate infection by zoonoses. The study evaluated the seroprevalence and risk factors associated with toxoplasmosis and toxocariasis in schoolchildren in the city of Jataizinho, Paraná. Of the 412 children aged 4-15 years, 56.8% (234/412) presented antibodies reactive to Toxoplasma gondii, 42.5% (175/412) presented antibodies reactive to Toxocara canis, and 27.4% (113/412) were reactive for the two species. The analysis of risk factors showed that prevalence of toxoplasmosis and toxocariasis was associated with the level of education of the child's mother (less than eight years of schooling), age range (10-15 years) and the presence of cats in the residence. In addition, family income (up to a minimum wage), presence of a dog, the habit of playing in soil/sand and eosinophilia were associated with Toxocara canis infection. There was an association between the two zoonoses (p < .01), indicating the existence of coinfection. The results show high prevalence of these two important zoonoses, alerting to the need of implementing control measures in order to reduce the incidence and risks of sequelae in children., (© 2021 Wiley-VCH GmbH.)

Background: Deficits in strength of the hip external rotators (ER) affect trunk, hip, and knee movement patterns, potentially contributing to injury in athletes., Objectives: To provide normative data on isometric torque for hip ER in athletes of three distinct sports and to determine if isometric torque for the hip ER and torque asymmetry between legs differ among sports and between sexes., Methods: Basketball, soccer, and volleyball athletes (n=451) were evaluated. Hip ER torque was quantified bilaterally with athletes in prone and 90° of knee flexion using a hand-held dynamometer., Results: Data are expressed as mean and 95% confidence interval. Hip ER torque values in Nm/kg for the dominant and non-dominant limbs were, respectively, 0.46 (0.44, 0.48) and 0.42 (0.40, 0.44) for male soccer athletes; 0.35 (0.32, 0.37) and 0.27 (0.25, 0.29) for male basketball athletes; and 0.37 (0.34, 0.39) and 0.35 (0.32, 0.37) for male volleyball athletes. Hip ER torque in Nm/kg for the female volleyball athletes was 0.29 (0.26, 0.33) for the dominant and 0.29 (0.25, 0.32) for the non-dominant limb. The Limb Symmetry Index for male soccer, basketball, and volleyball players was, respectively, 94% (91, 97), 81% (75, 87), and 95% (91, 99). For female volleyball players the Limb Asymmetry Index was 102% (95, 108). Male volleyball athletes showed higher torque values than female volleyball athletes., Conclusions: This study reported normative values for hip ER isometric torque of youth athletes. Clinicians can use the reported data as reference to identify torque deficits in athletes of the three reported sports., (Copyright © 2022 Associação Brasileira de Pesquisa e Pós-Graduação em Fisioterapia. Publicado por Elsevier España, S.L.U. All rights reserved.)

Cattle are an important source of zoonotic species of Cryptosporidium for humans. The aim of this study was to investigate the presence of Cryptosporidium, identify the species and determine the risk factors relating to environment, animals and management among dairy calves in eight Brazilian states. A total of 408 fecal samples from calves aged 1-60 days were analyzed. An epidemiological questionnaire was completed. Sample screening was performed using Ziehl-Neelsen technique and the positive samples were subjected to nested PCR. Cryptosporidium species were identified by means of the PCR-RFLP technique, using SSPI, ASEI and MBOII enzymes. The Ziehl-Neelsen technique showed that 89.7% (35/39) of the farms and 52.9% (216/408) of the samples were positive. Through nested PCR, these protozoa were detected in 54.6% of the samples. The 56 samples subjected to PCR-RFLP presented Cryptosporidium parvum. There was higher prevalence of the parasite in animals aged 7 to 28 days (62.6%). Diarrhea, ages between seven and 28 days and a spring water source were factors associated with the risk of infection. The calf hutch-type management system was associated with reduced infection. These findings demonstrate the high level of Cryptosporidium spp. circulation in cattle herds and the predominance of the species C. parvum.

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., (© 2021. Sociedade Brasileira de Microbiologia.)

Bees are insects of the order Hymenoptera and are involved in human accidents. In Brazil, bees that cause accidents are crosses derived from Europeans with African bees and are known for their aggressive behavior. Despite being considered an important public health concern, epidemiological studies at the national level are scarce. The objective of this study was to verify the epidemiological profile of bee accidents in humans in Brazil, using data from 2009 to 2019 of the Brazilian Ministry of Health. It was found that bee accidents increased by 207.61% from the first to the last year of the present study. The incidence varied according to the geographical region; the southern region had more bee accidents, but the Northern region had more deaths caused by bee accidents. Besides, climatic conditions were associated with susceptibility to bee stings; the incidence was higher during spring and summer. Age was also associated with fatality rate, with the elderly being the group with the highest fatality rate. Our results demonstrate that accidents caused by bees involve factors related to patients, the environment, and the behavior of bees. It is important to know the epidemiological aspects to help prevent apidic accidents., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

American cutaneous leishmaniasis (ACL) is a notifiable dermatozoonosis with relevant morbidity. The present study aimed to evaluate the epidemiological aspects of cases of ACL reported in Northern Paraná (2007-2016), as well as to georeference and analyse the influence of economic, social and environmental variables. Data from ACL notification forms were obtained and the patients' houses were georeferenced. Descriptive statistics, calculations of disease incidence and proportion of vegetation cover by municipality, spatial analysis, multiple linear regression and vulnerability analysis by census sector (CS) were performed. One thousand four hundred fifty-one cases of ACL were reported in the 89 municipalities of four regional health (RH). The average incidence of cases per 10,000 inhabitants was as follows: 11.58, 8.79, 4.92 and 4.03 in 18thRH, 15thRH, 16thRH and 17thRH. Peaks of incident cases were observed in 2008, 2012 and 2015. There was a statistically significant difference between the HR when comparing the proportions of the variables gender, age, education level, area of residence, clinical form, diagnostic criteria, response to treatment and drug used in case of failure. It was observed that the lower the proportion of remaining Atlantic Forest, the greater the incidence of ACL in the municipality. With regard to clusters analysis, treatment abandonment clusters were observed in the 15thRH and mucous form clusters were observed in the 15thRH, 16thRH and 17thRH. The vulnerability analysis by CS allowed us to observe a statistically significant difference in all vulnerability indicators: economic and social in 15thRH and 17thRH; home infrastructure in 15thRH and 18thRH, urban infrastructure in 15thRH and 18thRH and compound vulnerability index in 15thRH and 18thRH. However, the most vulnerable areas did not always have the highest number of cases. The data presented demonstrate that preventive and health education measures must be mainly directed to areas of greater degradation of native forest; regardless of the vulnerability situation., (© 2020 Wiley-VCH GmbH.)

The oocyst, a resistant form of Toxoplasma gondii, plays an important role in the transmission of this protozoan. The objective of this review was to report the methods capable of inactivating oocysts through a systematic review of the literature carried out in the Scientific Electronic Library Online, Web of Science, Science Direct, PubMed and Scopus databases. The keywords searched were (((effects OR infectivity OR resistance) AND Toxoplasma) AND oocyst). We selected 16 articles that described 309 different treatments. Among all the protocols evaluated, 35.60% (110/309) were effective in inactivating oocysts. Physical methods were more effective than other methods (p <0.05). Sporulated oocysts and the T. gondii VEG strain were more resistant (p <0.05) to treatments. Although it is effective against viruses and bacteria, the use of disinfectants in water has little or no effect on T. gondii oocysts. The use of radiation and pressure were effective in inactivating oocysts, as these treatments do not include changes in temperature, they can be used in foods for raw consumption, such as vegetables, as it will not cause substantially changes in their physical and chemical characteristics. Therefore, these methods can be viable alternatives for the control of T. gondii.

The aim of this study was to evaluate the seroprevalence and factors associated with the presence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Paraná state. The detection of antibodies for T. gondii and N. caninum was performed by homemade and commercial indirect ELISA, respectively. Multiple logistic regression analysis was used to verify the factors associated with the seroprevalence. Antibodies anti-T. gondii and anti-N. caninum were observed in 42.7% and in 17.6% of the animals, respectively. The protective factors associated to seropositive were "some level of confinement" (full or semi-extensive confinement) (OR=0.53) for T. gondii and "use of skilled labor" (OR=0.64) for N. caninum. The risk factors were "presence of cats" (OR=1.75) for T. gondii and "feeding of dogs with sheep placental remains" (OR=1.79) for N. caninum. In addition, to presenting a significant and simultaneous seroprevalence for both agents (9.9% of the animals), the results also indicate that deficiencies in management and environmental sanitation, the presence of reservoirs, and types of exploitation enhance the seropositivity. Thus, studies like this might support sanitary programs and public policies for the prevention of T. gondii and N. caninum in the sheep herds of Paraná state.

Although leptospirosis has been considered a major concern in urban areas, no study to date has spatially and simultaneously compared both owner and dog serology in households of major cities. Accordingly, the aim of the present study was to assess the seroprevalence of Leptospira antibodies, evaluate associated risk factors and conduct spatial analyses in 565 randomly selected households, which included 597 dog owners and 729 dogs in Londrina, Southern Brazil. Seropositivity by MAT were detected in in 11/597 (1.84%) owners and in 155/729 (21.26%) dogs. The risk factors were evaluated with logistic regression analysis and spatial factors and case distribution were evaluated with kernel density analyses. The sera of 14/155 (9.03%) dogs reacted for more than one serovar with the same titer. Canicola was the most frequent serogroup, detected in 3/11 (27.27%) owners and 76/155 (49.03%) dogs. The highest titer among the owners was 1:3,200 and was detected in the same household with a titer of 1:800 in the dog. Simultaneous owner-dog seropositivity was found in 7/565 (1.23%) households, with three reacted against serogroup Canicola. Positive owners were detected in 4/565 (0.70%) households and positive dogs were detected in 141/565 (24.95%) households. The associated risks of infection for dogs were different from those associated with infection in owners. Risk analyses for Canicola also identified specific factors of infection. Regardless of owner and dog cases were not statistically clustered, the kernel map has shown dog positivity occurrence in the same hot locations and near positive owners. The dependent variable analysis and logit model suggested a greater likelihood of peri-domiciliary contact with Leptospira . In conclusion, exposure to Leptospira infection was significantly higher in dogs than in their owners and human cases spatially overlapped dog cases, implicating dogs as potential environmental sentinels for this disease. In addition, the associated risk may vary according to serogroup, and the observed simultaneous Canicola seropositivity of owner and dog has suggested intradomicile-transmitted infection., Competing Interests: CM was employed by the company AAC&T Research Consulting LTDA. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Benitez, Monica, Miura, Romanelli, Giordano, Freire, Mitsuka-Breganó, Martins, Biondo, Serrano, Lopes, Reis, Gomes, Costa, Wunder, Ko and Navarro.)

Toxoplasmosis is a reportable disease in Brazil. The objective of this study was to investigate a toxoplasmosis outbreak at a research institution in Londrina-PR, Brazil. The outbreak was reported in October 2015; however, the first cases occurred in August 2015. Blood samples were collected from 674 persons at the institution. Samples were collected from soil, water (water tank) and food (vegetables) served in the restaurant. Each participant responded to an epidemiological questionnaire. For the blood samples, a chemiluminescent microparticle immunoassay was performed to detect IgM, IgG and specific IgG avidity antibodies; 10.8% (73/674) had evidence of acute toxoplasmosis. Statistical analysis showed a significant association (p < .001) between acute infection and eating lunch in the restaurant of the institution. Regarding the types of food offered in the restaurant during the period, there was a significant association between consuming raw salad (p < .001) and becoming ill. We conclude that the vegetables or raw vegetables served in the restaurant were probably the source of infection; however, the long period between exposure and case reporting made it difficult to identify the source of transmission., (© 2020 Blackwell Verlag GmbH.)

The present study aimed to describe a molecular analysis of environmental and pork samples, the isolation, genetic identification and immunohistochemistry (IHC) of Toxoplama gondii from placenta and amniotic fluid from five pregnant women that miscarried during a toxoplasmosis outbreak in 2018, Santa Maria, Rio Grande do Sul. Environmental and pork samples were submitted to polymerase chain reaction (PCR); placenta and amniotic fluid samples to histopathology, IHC, mouse bioassay and PCR. All samples were genotyped by PCR-RFLP with 11 loci. Histopathologic and IHC were compatibles with toxoplasmosis. All pregnants were positive in PCR and bioassay, the genotypes were compared, and all were equal suggesting a same source of infection. Among the environmental and food samples, a sludge sample from a water tank and two porks samples were positive in PCR, and the genotypes were different from the pregnant women isolates. It is concluded that obtain and compare isolates is essential to elucidate outbreak source., (Copyright © 2020 Elsevier B.V. All rights reserved.)

The aim of this study was to determine the occurrence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies and their associated factors among goats in farms from the Paraná state, South Brazil. The serological analysis was performed by indirect ELISA on 629 goat serum samples collected from 32 farms distributed in five mesorregions from the state. Seropositivity was observed in 30.7% of the animals for T. gondii, 6.3% for N. caninum, and 3.0% for both agents. Final multiple regression models showed that the use of assisted reproduction decreased the chance of seropositivity for T. gondii (PR=0.70) and the slaughtering site on the farm (PR=2.03) increased it. To N. caninum, the supplying concentrated feed to animals (OR=4.80) and animal pre-weaning death (OR=9.96) increased the chance of seropositivity. The variables associated with these seropositivities suggest deficiencies in sanitation and reproductive management. In addition, it recognizes critical points of infection for animal production management.

This study determined the prevalence of ovine toxoplasmosis and neosporosis and the risk factors associated with the development of these diseases in breeding rams from the State of Rio Grande do Sul (RS), Southern Brazil. Serum samples (n = 1,800) from breeding rams maintained on 705 sheep farms from seven mesoregions were evaluated serologically to detect anti-IgG Toxoplasma gondii by indirect ELISA and anti-IgG Neospora caninum by the indirect immunofluorescence assay (IFA). The prevalence of T. gondii was 33.05% (595/1,800); seropositivity to N. caninum was 18.44% (332/1,800). Additionally, there was simultaneous seropositivity (8.94%;161/1,800) to N. caninum and T. gondii. The variables size of the property (<500 ha) (Prevalence Ratio, PR = 1.36); breeding system (semi-intensive/intensive) (PR = 1.23); and natural mounting without control (PR = 1.50) were considered as risk factors for the occurrence of T. gondii. Size of the property (<500 ha) (PR = 1.58) and natural mounting without control (PR = 2.32) were risk factors associated with the prevalence of N. caninum in rams. Additionally, separation of ewes prior to parturition was considered as a protective factor for the occurrence of T. gondii (PR = 0.82) and N. caninum (PR = 0.74). These results demonstrated that these two parasitic disease agents are endemic in rams throughout all regions of RS., (© 2020 Blackwell Verlag GmbH.)

The present study attempted to verify the prevalence of and risk factors for diarrhea-causing agents in dairy calves from Brazil. Additionally, ages with a higher risk of occurrence for each agent were verified by means of the receiver operating characteristic (ROC) curve. The collections were performed on 39 farms, belonging to 29 municipalities located in eight states of Brazil. It was possible to conclude that the prevalence of Coronavirus, Rotavirus, Cryptosporidium spp., Eimeria spp., and nematodes was 7.20% (95% CI 4.54-9.78), 6.37% (95% CI 3.85-8.89), 51.52% (95% CI 45.26-55.57), 3.46% (95% CI 2.24-4.67), and 3.46% (95% CI 2.24-4.67), respectively. Ages with higher probabilities of occurrence of these diseases in calves were < 10, > 8, > 6, > 37, and > 36 days, respectively. Diarrhea occurred more significantly (P < 0.0001) in animals less than 21 days old and mainly on those receiving milk through automatic feeders (P < 0.001). Cryptosporidium spp. were a risk factor for the occurrence of Rotavirus, and vice versa (P = 0.0039) and presented a positive correlation with Coronavirus (P = 0.0089). Calves that drink water from rivers, streams, and ponds had a higher chance of being infected by Eimeria spp. (P < 0.0001), as well as developing infection by nematodes (P < 0.0001). The results found in this study highlight the importance of studying the agents of diarrhea together, once they act as coinfection where the losses triggered for the owners will involve some of these agents simultaneously.

Background: The objective of this study was to report an outbreak of human toxoplasmosis occurring in the municipality of Montes Claros de Goiás, Goiás, Brazil, from December 2015 to August 2016. Seven acute cases in June 2016 triggered the subsequent search., Methods: A total of 251 individuals were selected through an active search, of which 114 (45.4%) agreed to participate in the research and blood collection. For serological diagnosis were used enzyme-linked immunosorbent assay (ELISA) for IgG and IgM and avidity tests., Results: Of the 114 serum samples evaluated, 12.3% (14/114) showed antibodies against Toxoplasma gondii, with a profile indicative of acute infection. Samples of artisan fresh cheese, public water, vegetables and irrigation water were collected. Toxoplasma gondii DNA fragments were amplified using the polymerase chain reaction from two samples of artisan fresh cheese and a sample of irrigation water from the vegetable garden. A control case study was carried out, and the variable cow's artisan fresh cheese consumption was statistically significant (p = .01)., Conclusions: The results showed that cheese analysed and/or irrigated water of vegetable represented an important route of transmission for the disease. This is the first reported outbreak possibly caused by cow's artisan fresh cheese. It is difficult to prove that these routes were the cause of the outbreak; however, the findings allow us to infer that the individuals involved in the outbreak were in contact with these risk factors., (© 2019 Blackwell Verlag GmbH.)

Toxoplasma gondii is a protozoan that has great genetic diversity and is prevalent worldwide. In 2018, an outbreak of toxoplasmosis occurred in Santa Maria, Brazil, which was considered the largest outbreak ever described in the world. This paper describes the isolation and molecular characterization of Toxoplasma gondii from the placenta of two pregnant women with acute toxoplasmosis who had live births and were receiving treatment for toxoplasmosis during the outbreak. For this, placental tissue samples from two patients underwent isolation by mice bioassay, conventional PCR and genotyping using PCR-RFLP with twelve markers. Both samples were positive in isolation in mice. The isolate was lethal to mice, suggesting high virulence. In addition, the samples were positive in conventional PCR and isolates submitted to PCR-RFLP genotyping presented an atypical genotype, which had never been described before. This research contributes to the elucidation of this great outbreak in Brazil., Competing Interests: The authors have declared that no competing interests exist.

The aim of this study was to perform a systematic review of scientific papers that used spatial analysis tools in cases of leishmaniasis, in Brazil. The search for articles was carried out in PubMed, SciELO, Scopus and Web of Science databases. The keywords used in the identification of the articles were Thematic map AND Leishmaniasis, Spatial analysis AND Leishmaniasis, and Geoprocessing AND Leishmaniasis, in English language. A total of 360 articles were found, and 11 of them were analyzed after screening by title and abstract as well as reading of the full articles. The States studied were Sao Paulo, Acre, Maranhao, Piaui, Minas Gerais, Parana and Tocantins. Cutaneous leishmaniasis occurred predominantly in rural areas, with clusters in forest reserve regions or modified forest areas. Conversely, visceral leishmaniasis mainly occurred in peripheral and central urban areas associated with poorer environments and urban infrastructure, including worse sanitation. We conclude that the spatial distribution of leishmaniasis is closely related to the living environment of the risk population. The analyzed articles associated geospatial data with some risk factors for the disease, pointing out the locations where most cases occur, creating a relevant source to define control strategies.

We report on apparent temporal progression of probable sources of infection and transmission routes for global human toxoplasmosis outbreaks as described in published articles. We searched the Scientific Electronic Library Online, Web of Science, PubMed, and Scopus databases for articles on Toxoplasma, toxoplasmosis, and outbreaks. We found that transmission routes for Toxoplasma gondii varied by decade. In the 1960s and 1990s, toxoplasmosis outbreaks mainly occurred through ingestion of cysts in meat and meat derivatives; in the 1980s, through milk contaminated with tachyzoites; in 2000, due to the presence of oocysts in water, sand, and soil; and in 2010, due to oocysts in raw fruits and vegetables. Our study suggests a possible change in the epidemiology of reported toxoplasmosis outbreaks. Because of this change, we suggest that greater attention be paid to the disinfection of vegetables, as well as to the quality of water used for drinking and irrigation.

The aim of this study was to confirm the emergence of canine visceral leishmaniasis among dogs in Foz do Iguaçu. The disease was diagnosed through the isolation and molecular identification of Leishmania infantum. In the first sample collection stage (2012), three lymph node aspirates and 46 buffy coat samples were obtained mostly from the dogs that were seroreagents for leishmaniasis. In the second sample collection stage (2013), the buffy coat samples were collected from 376 dogs located close to Paraguay, Paraná river, center and peripheral parts of the city. The DNA from the six isolates, four from the first sampling stage (4/49) and two from the second sampling stage (2/376), was subjected to polymerase chain reaction using the K26F/R primers. The isolate was confirmed as L. infantum by sequencing. As none of the dogs had ever left the city, the isolates were confirmed as autochthonous. Further, the study confirmed the emergence of canine visceral leishmaniasis in Paraná through the identification of L. infantum among dogs in Foz do Iguaçu city. Hence, collaborative control measures should be designed and implemented by the public agencies and research institutions of Brazil, Argentina, and Paraguay to control the spread of visceral leishmaniasis.

The objective of this study was to report an outbreak of human toxoplasmosis at a research institution in Londrina, Paraná, from December 2015 to February 2016. Blood samples from 26 symptomatic individuals were collected and the microparticle chemiluminescence immunoassay was performed to detect IgM, IgG and specific IgG avidity test in the official laboratory. A total of 20 people with symptoms and serology compatible with acute toxoplasmosis (IgM positive and IgG with low avidity) were selected as cases, while 45 asymptomatic employees working in the same teams and during the same shifts were selected as controls. All the participants of the investigation answered an epidemiological questionnaire. Three samples of water and one sludge from the institution's supply cisterns, 10 soil samples, 11 plant samples, three cat fecal samples and one domestic feline cadaver were collected for analysis of the polymerase chain reaction (PCR) for T. gondii. After analyzing the epidemiological data, the consumption of vegetables in the restaurant of the institution was the only variable associated with the occurrence of the disease. In laboratory results, all the samples showed negative results to PCR. The rapid recognition of the outbreak, early notification and investigation could have broken the chain of transmission early, thus preventing the emergence of new cases. In addition, the adoption of good food handling practices could have prevented the occurrence of the outbreak.

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., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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.

Human toxoplasmosis, a protozoonosis caused by Toxoplasma gondii, has been described as a worldwide foodborne disease with important public health impact. Despite infection has reportedly varied due to differences in alimentary, cultural and hygienic habits and geographic region, social vulnerability influence on toxoplasmosis distribution remains to be fully established. Accordingly, the present study has aimed to assess T. gondii seroprevalence and factors associated to social vulnerability for infection in households of Ivaiporã, southern Brazil, with 33.6% population making half minimum wage or less, ranked 1,055th in population (31,816 habitants), 1,406th in per capita income (U$ 211.80 per month) and 1,021st in HDI (0.764) out of 5,570 Brazilian cities. Serum samples and epidemiological questionnaires were obtained from citizen volunteers with official City Secretary of Health assistance in 2015 and 2016. In overall, serosurvey has revealed 526/715 (73.57%) positive samples for anti-T. gondii antibodies by Indirect Fluorescent Antibody Test. Logistic regression has shown a significant increase associated to adults (p = 0.021) and elderly (p = 0.014) people, illiterates (p = 0.025), unemployment (p <0.001) and lack of household water tank (p = 0.039). On the other hand, sex (male or female), living area (urban or rural), yard hygiene, meat ingestion, sand or land contact, owning pets (dog, cat or both) were not significant variables of positivity for anti-T. gondii antibodies in the surveyed population. Although no significant spatial cluster was found, high intensity areas of seropositive individuals were located in the Kernel map where the suburban neighborhoods are located. In conclusion, socioeconomic vulnerability determinants may be associated to Toxoplasma gondii exposure. The increased risk due to illiteracy, adult or elderly age, unemployment and lack of household water tank were confirmed by multivariate analysis and the influence of low family income for seropositivity by the spatial analysis., Competing Interests: The authors have declared that no competing interests exist.

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.

The aim of this study was to investigate the occurrence of anti-Leishmania spp. antibodies in dogs from localities in the city of Foz do Iguaçu, Paraná state, Brazil, on the border with Argentina and Paraguay. Blood samples dogs were collected to perform the following serologic tests: immunochromatographic DPP® rapid test, indirect immunoenzymatic assay (ELISA) and indirect immunofluorescence assay (IFA). In 2012, 285 dogs were analyzed on Argentina border, and in 2013, serum samples from 396 dogs on the border of Paraguay were collected. Using ELISA for screening and IFA for the confirmatory test, the results showed that the antibody prevalence was 1.8% (5/285) on the border of Argentina and 3.0% (12/396) on Paraguay border. When using the DPP® for screening and ELISA as a confirmatory analysis, we observed a seroreagent prevalence in dogs of 2.5% (7/285) on Argentina border and 5.1% (20/396) on Paraguay border. The non-public collection of domestic waste (p= 0.0004) was shown to be associated with leishmaniasis. This study shows the presence of leishmaniasis and suggest the emergence of canine visceral leishmaniasis in state of Paraná due to the confirmed occurrence of seroreactive dogs on Argentina and Paraguay border, which has environmental and geographical characteristics that favor the spread of the parasite.

The objective of this study was to determine factors associated with vegetable contamination with zoonotic protozoan. Samples of water, soil and vegetables were collected from July/2014 to May/2016, totaling 83 samples, 21 properties of Londrina region, Paraná, Brazil. DNA amplification of Toxoplasma gondii, Cryptosporidium spp. and Giardia intestinalis in the samples was conducted using polymerase chain reaction (PCR). The PCR results were positive for T. gondii in 12.9% (8/62), Cryptosporidium spp. in 11.3% (7/62) and G. intestinalis in 25.8% (16/62) of the samples. DNA sequencing identified C. parvum in five samples and G. intestinalis Assemblage E in three. The statistical associations demonstrated greater probability of positive samples for T. gondii and for at least one of the three protozoa when the source of irrigation water was the river; a greater chance of positive samples for Cryptosporidium spp. when deer were present on the property; and a smaller chance of positive samples for at least one of the three etiologic agents when soil was supplemented with limestone. The results expose some critical contamination points, providing support for training farmers on good management practices during the production process.

Although leishmaniasis has been described as a classic example of a zoonosis requiring a comprehensive approach for control, to date, no study has been conducted on the spatial distribution of simultaneous Leishmania spp. seroprevalence in dog owners and dogs from randomly selected households in urban settings. Accordingly, the present study aimed to simultaneously identify the seroprevalence, spatial distribution and associated factors of infection with Leishmania spp. in dog owners and their dogs in the city of Londrina, a county seat in southern Brazil with a population of half a million people and ranked 18th in population and 145th in the human development index (HDI) out of 5570 Brazilian cities. Overall, 564 households were surveyed and included 597 homeowners and their 729 dogs. Anti-Leishmania spp. antibodies were detected by ELISA in 9/597 (1.50%) dog owners and in 32/729 (4.38%) dogs, with significantly higher prevalence (p = 0.0042) in dogs. Spatial analysis revealed associations between seropositive dogs and households located up to 500 m from the local railway. No clusters were found for either owner or dog case distributions. In summary, the seroepidemiological and spatial results collectively show a lack of association of the factors for infection, and the results demonstrated higher exposure for dogs than their owners. However, railway areas may provide favorable conditions for the maintenance of infected phlebotomines, thereby causing infection in nearby domiciled dogs. In such an urban scenario, local sanitary barriers should be focused on the terrestrial routes of people and surrounding areas, particularly railways, via continuous vector surveillance and identification of phlebotomines infected by Leishmania spp., (Copyright © 2018. Published by Elsevier B.V.)

The aim of this study were to detect antibodies anti-Leishmania spp. and anti-Trypanosoma cruzi in two different populations of domestic cats (Felis catus domesticus) from North Paraná referred for surgical castration and to determine which characteristics of the animals studied may be associated with seropositivity. Serum samples from 679 cats were analyzed using enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence antibody test (IFAT) in series. Associations between age, sex, race, year of care and animal group were verified using the simple logistic regression. Percentage of 8.5% (58/679) of cats were positive for Leishmania spp. and 7.6% (51/673) for T. cruzi by the tests ELISA and IFAT. Animals collected by non-governmental animal protection organizations presented more seropositivity for Leishmania spp. (p<0.0001). Results shown that Leishmania spp. and T. cruzi are present in domestic cats in the northern part of the state of Paraná, as well as, owners of non-governmental animal protection organizations may be more exposed to leishmaniasis when compared to other animal owners evaluated in the present study.

[This corrects the article DOI: 10.1371/journal.pone.0180906.].

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.

Rearing free-range chicken is based on grazing feeding patterns, and these animals could be potential environmental contaminants of Cryptosporidium oocysts for humans and other animals. Therefore, the present study aimed to evaluate the molecular prevalence of Cryptosporidium spp. in free-range chickens from Brazil. A total of 351 fecal samples from chickens were examined from 20 farms. For detection of Cryptosporidium spp., 18S rRNA gene fragments were amplified using a nested PCR reaction. Positive samples were sent for sequencing. The overall prevalence of Cryptosporidium was 25.6% (95% CI = 21.2% - 30.6%). Sequencing of the amplified fragments allowed for the identification of three species: C. meleagridis in 57 (62.6%), C. baileyi in 15 (16.4%), C. parvum in 3 (3.2%) samples, and a new Cryptosporidium genotype (C. genotype BrPR1) in 3 (3.2%) samples. Cryptosporidium genotype BrPR1 has not yet been classified as a species, and its host spectrum is not known. Cryptosporidium, including zoonotic species, exists at a high prevalence in free-range chickens within the region studied.

Toxoplasmosis, caused by Toxoplasma gondii, has traditionally been considered an important water and foodborne protozoonosis with important public health considerations. Although felids play a well-established role as definitive hosts, canine epidemiological involvement in the parasite's life cycle remains questionable and controversial. The increasing closeness of the human-dog bond, particularly seen in urban settings, has been recognized as a historically unprecedented worldwide movement. Sharing daily lives in the same households, dogs may be exposed to similar associated risks of T. gondii infection as their owners. Thus, epidemiological assessment of the intra-domiciled environment, especially among socio-economically different human populations, may provide novel information regarding the actual role of dogs in animal and human toxoplasmosis. Despite spatial approaches being recently used for other water and foodborne diseases, no study has been conducted on the simultaneous spatial seroprevalence of both human and animal IgG anti-T. gondii antibodies in urban areas of major cities. Accordingly, the aim of the present study was to assess the seroprevalence and associated variables of Toxoplasma infection in owners and their domiciled dogs in Londrina, southern Brazil. Human and canine seroprevalence rates and variables associated with seroprevalence were investigated through representative random sampling among 564 households, which included 597 owners and 729 dogs. Overall, statistically significant differences between the seroprevalence of human and dog anti-T. gondii antibodies were found by Immunofluorescence Antibody Testing in 248/597 (41.54%) owners and 119/729 (16.32%) dogs. Through multiple analysis, significant concomitant variables for seropositivity of household individuals (people and dogs) were determined, including public sewer service, yard cleaning frequency, and having a dirty yard. Although no statistically significant multiple logistic model was observed among owners, univariate analysis detected associations with monthly income, soil contact, and occupation. Among dogs, the absence of other dogs and the absence of a dirty yard were concomitant significantly protective associated factors. Age differences between seropositive and seronegative individuals was significant only for human beings, with the median age of negative individuals significantly higher than positive individuals. Although no spatial clusters were identified for humans or residences, a significant cluster was identified for dogs. In conclusion, characteristics of urban toxoplasmosis may include significantly higher owner seroprevalence than their owned dogs, with canine seroprevalence directly associated with having more dogs and a dirty backyard, and spatial differences in both human and dog exposures. Although not a good indicator for human foodborne diseases, dogs may be a reliable sentinel for environmental infection. Moreover, such a holistic approach may provide crucial information for more focused prevention and monitoring programs, particularly in households with multiple pets and trash-filled backyards.

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.

Although the infection by Toxoplasma gondii in horses is usually asymptomatic, it may be characterized by the presence of tissue cysts and maintenance of detectable antibody titers. The aim of this study was to detect anti-T.gondii antibodies and to determine risk factors associated in sport horses from Paraíba State, Northeastern Brazil. A total of 138 apparently healthy sport horses of different age and gender were sampled. Anti-Toxoplasma gondii antibodies were detected in 16/138 (11.6%; 95% CI: 6.77-18.14%) horses by indirect immunofluorescence antibody assay (IFA). A significant association was found between the water source and seropositivity to T. gondii (p<0.05). The present study highlights the importance of providing good quality water to the animals.

This study aimed to detect parasites from Leishmania genus, to determine the prevalence of anti-Leishmania spp. antibodies, to identify circulating species of the parasite, and to determine epidemiological variables associated with infection in rats caught in urban area of Londrina, Paraná, Brazil. Animal capture was carried out from May to December 2006, serological and molecular methods were performed. DNA was extracted from total blood, and nested-PCR, targeting SSu rRNA from Leishmania genus, was performed in triplicate. The positive samples were sequenced twice by Sanger method to species determination. In total, 181 rodents were captured, all were identified as Rattus rattus and none showed clinical alterations. Forty-one of the 176 (23.3%) animals were positive for Leishmania by ELISA and 6/181 (3.3%) were positive by IFAT. Nine of 127 tested animals (7.1%) were positive by PCR; seven were identified as L. (L.) amazonensis, one as L. (L.) infantum. Four rats were positive using more than one test. This was the first description of synanthropic rodents naturally infected by L. (L.) amazonensis (in the world) and by L. (L.) infantum (in South Brazil). Regarding L. (L.) amazonensis, this finding provides new evidence of the urbanization of this etiological agent.

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.

We report the results of extensive computational investigation of the adsorption properties of water and ethanol on several Cu-, Pt-, and Au-based substrates, including the close-packed unreconstructed Cu(111), Pt(111), and Au(111) surfaces, defected metal substrates with on-surface low-coordinated sites generated by the intermixing of Pt-Cu and Pt-Au in the topmost surface layers and strained on-surface and sub-surface Pt-layers at Cu(111) and Au(111) substrates. The calculations are based on the density functional theory (DFT) within the van der Waals (vdW) correction. For all the substrates, we found that water and ethanol bind via the anionic O atom to the cationic one-fold coordinated on-top metal sites, which enhances the adsorbate-substrate Coulomb interactions. For water, both DFT and DFT + vdW calculations predict a flat geometry. For ethanol, the DFT and DFT + vdW results are in contrast, namely, DFT yields a perpendicular orientation of the C-C bond with respect to the surface, while we obtained a parallel orientation of the C-C bond using DFT + vdW, which maximizes the adsorption energies. Despite expected deviations due to the nature of the weak adsorbate-substrate interactions, we found that the adsorption energy of water and ethanol shows a linear dependence as a function of the position of the center of gravity of the occupied d-band, and hence, the magnitude of the adsorption energy increases as the d-band center position shifts towards the Fermi energy. Thus, it indicates hybridization between the O p- and metal d-states, which determines the magnitude of the adsorption energy of water and ethanol on clean, low-coordinated, and strained noble and transition-metal substrates.

Background: The Brazilian exportation of pet food has shown high growth rates in the last two years and determination of the exposure degree is one of the most important parameters for the risk assessment of chemical compounds. In this study the exposure degree of dogs to mycotoxins was estimated and acceptable daily intake (ADI) and safe pet dietary level (SPDL) were calculated. Thus the natural occurrence of fumonisins, zearalenone and aflatoxins was evaluated in 100 dry dog feed samples provided by pet owners in Paraná State, Brazil., Results: Despite the high frequency of fumonisins (68%), zearalenone (95%) and aflatoxins (68%) in feed samples, the mean levels detected were low. ADI for fumonisins and zearalenone was 20.0 and 1.00 µg kg(-1) body weight (BW) day(-1) respectively and SPDL for fumonisins was 2000 µg kg(-1) feed. The probable daily intake values (1.83 µg fumonisins, 0.93 µg zearalenone and 0.02 µg aflatoxins kg(-1) BW day(-1) ) were low., Conclusion: The exposure degree of dogs could be assumed to be very low. However, the co-occurrence of these three or other mycotoxins, and possible synergic or additive effects, should be taken into account when determining the maximum allowed levels or risk assessment. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

The role of rodents in the epidemiology of toxoplasmosis was investigated in Londrina, Paraná State, Brazil. One hundred and eighty-one Rattus rattus and one Mus musculus were caught in 37 places. Blood and tissues were collected and submitted to the indirect fluorescence antibody test (IFAT) and the bioassay. Serum samples from 61 contacting dogs were also collected. Sixteen rats (8.8%) were positive for Toxoplasma gondii, but just two of them were positive by serology and bioassay test. Antibodies were found in nine (4.9%) rats. Tissues of nine rats bioassayed were positive and four isolates were obtained. Restriction fragment length polymorphism (RFLP) analysis was performed using 12 markers (SAG1, SAG2, SAG2-alt, C22-8, C29-2, L358, PK1, BTUB, GRA6, SAG3, Apico, CS3). Genotyping revealed that the four strains isolated from this study have been isolated before in cats and chickens from Brazil. None of the isolates was identified like clonal archetypal T-types I, II, and III. The rats presented lower serologic Toxoplasma gondii prevalence (8.8%) compared to contacting dogs (70.5%).

The aim of this study was to determine the presence of mycotoxins on dogs feed and to explore the potential association between mycotoxins exposure and the chance of mamary tumors in a case-control study. The study included 256 female dogs from a hospital population, 85 with mammary tumors (case group) and 171 without mammary tumors (control group). An epidemiological questionnaire was applied to both groups, and the data were analyzed by the EpiInfo statistical package. For the study, 168 samples of the feed offered to dogs were analyzed for the presence of aflatoxins, fumonisins and zearalenone by high-performance liquid chromatography. Mycotoxins were found in 79 samples (100%) in the case group and 87/89 (97.8%) in the control group. Mycotoxins were detected in all types of feed, regardless feed quality. Level of aflatoxin B1 (p = 0.0356, OR = 2.74, 95%, CI 1.13 to 6.60), aflatoxin G1 (AFG1) (p = 0.00007, OR = 4.60, 95%, CI = 2.16 to 9.79), and aflatoxin G2 (AFG2) (p = 0.0133, OR = 9.91, 95%, CI 1.21 to 81.15) were statistically higher in case of mammary cancer. In contrast, neutering was a protective factor for mammary cancer (p = 0.0004, OR = 0.32, 95%, CI = 0.17 to 0.60)., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

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.

Despite extensive studies of transition metal (TM) clusters supported on ceria (CeO2), fundamental issues such as the role of the TM atoms in the change in the oxidation state of Ce atoms are still not well understood. In this work, we report a theoretical investigation based on static and ab initio molecular dynamics density functional theory calculations of the interaction of 13-atom TM clusters (TM = Pd, Ag, Pt, Au) with the unreduced CeO2(111) surface represented by a large surface unit cell and employing Hubbard corrections for the strong on-site Coulomb correlation in the Ce f-electrons. We found that the TM13 clusters form pyramidal-like structures on CeO2(111) in the lowest energy configurations with the following stacking sequence, TM/TM4/TM8/CeO2(111), while TM13 adopts two-dimensional structures at high energy structures. TM13 induces a change in the oxidation state of few Ce atoms (3 of 16) located in the topmost Ce layer from Ce(IV) (itinerant Ce f-states) to Ce(III) (localized Ce f-states). There is a charge flow from the TM atoms to the CeO2(111) surface, which can be explained by the electronegativity difference between the TM (Pd, Ag, Pt, Au) and O atoms, however, the charge is not uniformly distributed on the topmost O layer due to the pressure induced by the TM13 clusters on the underlying O ions, which yields a decrease in the ionic charge of the O ions located below the cluster and an increase in the remaining O ions. Due to the charge flow mainly from the TM8-layer to the topmost O-layer, the charge cannot flow from the Ce(IV) atoms to the O atoms with the same magnitude as in the clean CeO2(111) surface. Consequently, the effective cationic charge decreases mainly for the Ce atoms that have a bond with the O atoms not located below the cluster, and hence, those Ce atoms change their oxidation state from IV to III. This increases the size of the Ce(III) compared with the Ce(IV) cations, which builds-in a strain within the topmost Ce layer, and hence, also affecting the location of the Ce(III) cations and the structure of the TM13 clusters.

The objective of the present study was to evaluate infection of cats by Paracoccidioides brasiliensis. Serum samples of 136 cats from rural (n = 86) and urban areas (n = 50) were analyzed by indirect ELISA and immunodiffusion test using P. brasiliensis gp43 and exoantigen as antigens, respectively, and an overall reactivity of 31.6 % was observed by ELISA although no reactivity was detected by immunodiffusion. The positivity observed in animals living in rural areas (48.8 %) with free access to soil was significantly higher (P < 0.0001) than among urban animals (2 %) with limited access to soil, although no significant difference was observed in relation to age or sex. The high rates of positivity observed in cats from rural areas suggest that not diagnosed cases of this mycosis may be occurring in cats living in endemic areas for human paracoccidioidomycosis. This is the first report showing serological evidence of P. brasiliensis infection in cats.

The objective was to characterize the transmission of Toxoplasma gondii in goats experimentally infected vaginally with semen contaminated with the CPG strain (genotype III). Ten female goats were randomly allocated into 2 groups (G1 and G2), each with 5 animals, and inseminated during estrus. Goats in G1 were inseminated with semen containing 1 × 10(5) tachyzoites, whereas those in G2 (control) were inseminated with semen free from tachyzoites (insemination = day 0). In G1, seroconversion (indirect immunofluorescence reaction) and DNA (polymerase chain reaction) in the blood was present in 4/5 and 3/5, respectively, from the 7th day. In G2, all goats were negative in all tests. Embryonic reabsorption occurred in 4 of 5 goats from G1 between days 21 and 49. In conclusion, artificial vaginal insemination with semen containing tachyzoites of T. gondii -infected goats and is a potential transmission route of this parasite through semen.