Search

In Brazil, the wild boar (Sus scrofa) is the main invasive exotic species, responsible for causing various economic, environmental, and health damages to biodiversity. In view of the above, we aimed in our study to investigate the occurrence of endo and ectoparasites in free-ranging wild boars in the northwest region of the state of São Paulo, Brazil. In total, 45 animals of different sexes and ages were examined in this research. Fecal and urine samples were processed using coproparasitological techniques. Multiple incisions in muscles, liver, and pancreas were also made to investigate cestodes and trematodes, and the entire epidermal region of the animals was analyzed for ectoparasites search. In our study, we demonstrate an occurrence of 97.77% (44/45) and 43.33% (13/30) of endo and ectoparasites, respectively, in wild boars, with the identification of eggs of Metastrongylus spp., Strongyloides ransomi, Trichuris suis, Stephanurus dentatus, and nematodes of the order Strongylida, as well as oocysts of Eimeriidae protozoa. Ectoparasite infestations were mainly caused by ixodids of the genus Amblyomma spp. and by larvae of Cochliomyia hominivorax., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

This study assessed the seroprevalence of Toxoplasma gondii and risk factors among 428 pregnant women attending Basic Health Units (BHUs) in Araçatuba, São Paulo, Brazil. The seroprevalence was 55.14%, indicating high exposure to the parasite in this population. Using a multi-level logistic regression model, this study analyzed these predictors to determine their association with a higher seropositivity rate, with BHUs included as a random factor. Predictors associated with higher seropositivity included older age (36-45 years), with a 71.64% prevalence in this group, and multiparity (61.65%). Women with lower educational levels were also more likely to be infected, with 59.46% seropositivity recorded among those who had only completed elementary school. Despite identifying several risk factors, no significant correlation was found between undercooked meat consumption or contact with soil and infection. These findings highlight the need for targeted public health interventions, particularly for educating high-risk groups about toxoplasmosis prevention, such as safe food handling and avoiding raw dairy products. Additionally, BHUs play a critical role in early detection and prevention. These units are important for providing healthcare access and preventive education for vulnerable populations. Given the high seroprevalence, this study underscores the urgency of implementing prenatal screening and educational programs to reduce the risks of congenital toxoplasmosis in this region.

This study examined the seropositivity of T. gondii and coinfections with other TORCH pathogens among pregnant women attending 17 Basic Health Units (UBS) in Araçatuba, SP, Brazil. Of the 711 pregnant women seen at these UBS, only 297 were tested for T. gondii . Of the women tested for T. gondii ( n = 297), 26.9% had IgG antibodies, 6.7% had IgM, and 32.0% tested positive for either or both. Only 1.4% showed both IgG and IgM antibodies, while 67.7% were non-reactive. The seropositivity was 17.1% for syphilis, 63.2% for rubella, 0.9% for hepatitis C, 0.9% for dengue, 17.9% for COVID-19, and 0.9% for herpes simplex (types 1/2). Coinfections with syphilis, rubella, and herpes simplex were also noted. Higher education levels appeared to protect against T. gondii seropositivity. The findings highlight a significant prevalence of T. gondii among pregnant women, with variation across UBSs, pointing to socioeconomic, behavioral, and environmental factors as influential. We also observed co-occurrence with other infections, such as syphilis, rubella, and herpes simplex. The study underscores the need for targeted public health interventions to reduce the risks of congenital infections.

Although previous studies have characterized the helminth fauna of wild boars kept in captivity in Brazil, records on these helminths in free-ranging animals are still scarce. In view of this, we aimed in our work to investigate the occurrence and morphological and morphometric characteristics of gastrointestinal helminths in wild Sus scrofa from the northwest region of the State of São Paulo, Brazil. The digestive systems of 10 animals (5 males and 5 females of different ages) were used in this study. Each anatomical segment was washed and sieved under running water, and the helminths were separated and identified using light and scanning electron microscopy, according to their morphological characteristics. A total of 2750 (1152 males and 1598 females) nematode specimens were collected from the small intestine of these wild boars, and all of them presented the morphological characteristics of Globocephalus urosubulatus . However, one characteristic is of particular interest because it has not yet been reported in the literature: a marked asymmetry between the lobes and their respective rays of the copulatory bursa, with the left one being larger than the right one. In this research, we identified the presence of G. urosubulatus in all the examined free-ranging wild boars and reported for the first time in the literature the asymmetry in the copulatory bursa.

Visceral leishmaniasis is a zoonotic disease that affects humans and dogs. The infection is endemic in the municipality of Araçatuba, São Paulo, Brazil. Given the role of dogs in the epidemiology of leishmaniasis, strategies to enhance surveillance and reduce transmission are focused on dogs. In this study, we retrospectively analyzed records of canine visceral leishmaniasis from 2013 to 2022. According to this database, the prevalence of dogs testing positive for leishmaniasis fluctuated, with an average of 65.04% (6590/10,133). Cases were clustered in 10 statistically significant areas. Environmental analyses identified a significant geographical association between animals testing positive and higher vegetation density rates compared with animals testing negative. The period from sample collection to diagnosis and euthanasia, as recommended by the Brazilian Ministry, correlated with disease prevalence and decreased over time. These findings serve to implement different action plans against leishmaniasis for each geographic region and to understand the impact and efforts of strategies in an endemic area.

Background: Discarding pharmaceuticals in the garbage or into the sewage system are still the most common methods in many countries. This study aims to investigate the guidance provided by pharmacists to customers on the disposal of unused and expired household medications in São Paulo State, Brazil., Method: The study population consisted of 630 pharmacists from the State of São Paulo, who work in community pharmacies. They answered an online questionnaire with questions composed in three blocks: demographic, work, and academic information on the pharmacist; guidance about the disposal of household medications; and knowledge regarding the reverse logistics of these medications. An invitation to participate in the questionnaire was made via WhatsApp, individually and collectively. Inferential statistics were performed using the chi-square test and were considered significant when p < 0.05%., Results: Among the participating pharmacists, the majority were women under 60 years old,56 (8.89%) stated that they never orient the customer regarding the disposal of unused and expired household medications, while 574 (91,12%) indicated that they almost provide guidance. The frequency with which they provided guidance was influenced by the number of years since graduation (p = 0.0047), the time they had worked in pharmacies and drugstores (p = 0.0007), and whether or not they had a graduate degree (p = 0.0181). Regarding the disposal of medications, among the 643 responses provided by the pharmacists,516 (80.25%) indicated that they oriented customers to return them to a pharmacy., Conclusion: A small number of pharmacists always orient customers on the proper disposal that should be followed for unused and expired household medications, prioritizing their return to a pharmacy. In general, these pharmacists have longer periods of work experience and higher academic qualifications. Thus, it is important to increase knowledge through professional training and further education programs., (© 2023. The Author(s).)

We investigated the zoonotic transmission of Cryptosporidium among the children ( n = 188), dogs ( n = 133), and cats ( n = 55) living in 188 households. Fecal samples were examined using ELISA and confirmed via nested PCR. Coproantigens oocysts were detected in 3.7% of children, 8.3% of dogs, and 5.5% of cats. We found strong evidence of two cases of the zoonotic transmission of Cryptosporidium canis between children and dogs. Furthermore, four children and their respective pets (one dog and three cats) were infected with Cryptosporidium parvum , but we cannot exclude the hypotheses that the oocysts were transmitted from children to animals or that both hosts were infected by a shared source, such as contaminated water or food. The presence of an infected animal elevated the risk of zoonotic transmission by 129.7-fold (95% CI: 13.92-1209.68). Furthermore, sharing a bed with pets was identified as a risk factor for infection in children (OR: 9.9, 95% CI: 1.37-71.2). In conclusion, the zoonotic transmission of Cryptosporidium among children and pets cohabiting in the same household may be quite common, especially when infected animals lie or sleep on children's beds. These findings unequivocally highlight the public health concern surrounding C. canis .

We investigated the occurrence of gastrointestinal parasites in dogs in rural areas of the municipality of Painel, Santa Catarina, Brazil. For this, 91 canine feces samples were collected from 82 farms between August, 2017 and January, 2018. These fecal materials were processed using the techniques of spontaneous sedimentation, centrifugation-floatation in zinc sulfate and Ziehl-Neelsen staining. A questionnaire in the form of an interview was administered to the dogs' owner and information about the farm and the main care provided for the dogs was obtained. Among 91 sampled dogs, 27 (29.7%) were positive for at least one parasite species. Ancylostoma was the most prevalent genus (16.5%), followed by Giardia duodenalis (14.3%), Trichuris vulpis (6.6%), Toxocara canis (5.5%), Entamoeba spp. (4.8%), Cryptosporidium spp. (3.3%) and Taeniidae (1.1%). Most dogs do not receive veterinarian care and rarely received antiparasitic treatment. They were free to roam and had free access to animal remains and garbage, which was reflected in the significant associations with the occurrence of parasites that were found. We conclude that rural dogs harbor gastrointestinal parasites, but that their owners are unaware of the risks that these parasites can bring to human health.

Giardiasis is a major cause of diarrhea in humans and animals worldwide. Currently, there are nine species of Giardia, including Giardia duodenalis, which infects most vertebrates. The capybara (Hydrochoerus hydrochaeris) is the largest herbivorous rodent in the world. Although capybaras are hosts of several parasites of public health importance, including helminths and protozoa, there is a paucity of research on their zoonotic potential. We investigated the prevalence of Giardia spp. in populations of capybaras living in urban areas. Fecal samples from 247 capybaras were collected in Lagoa Maior, located in the municipality of Três Lagoas, and in Lago do Amor and Parque das Nações Indígenas, both located in the municipality of Campo Grande, state of Mato Grosso do Sul, Brazil. Fecal samples from capybaras originated from 133 adults (54%), 61 cubs (25%), and 53 juveniles (21%); 183 samples were collected in the rainy season and 64 in the dry season. Giardia spp. DNA was screened by the small-subunit ribosomal RNA (SSU rRNA) targeted PCR. Samples with DNA band sizes suggestive of Giardia spp. amplicons were examined by PCR targeting the glutamate dehydrogenase (GDH) and triosephosphate isomerase (TPI) genes. PCR amplicons were subjected to genetic sequencing. Nested PCR screening of the SSU rRNA gene revealed 16 samples showing faint DNA bands in gel electrophoresis with sizes similar to Giardia spp. amplicons. PCR amplicons of the SSU rRNA gene were analyzed by Sanger sequencing. Most of the sequencing reactions failed, and the chromatogram reads of some samples were ambiguous, suggesting nonspecific amplification. Therefore, all the capybara fecal samples were considered negative for Giardia spp. Two published studies on Giardia spp. in capybaras reported findings similar to ours, i.e., the absence or a low positivity rate for Giardia spp. However, further studies are needed to determine the possible role of capybaras in the epidemiology of giardiasis., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Diagnosing gastrointestinal parasites by microscopy slide examination often leads to human interpretation errors, which may occur due to fatigue, lack of training and infrastructure, presence of artifacts (e.g., various types of cells, algae, yeasts), and other reasons. We have investigated the stages in automating the process to cope with the interpretation errors. This work presents advances in two stages focused on gastrointestinal parasites of cats and dogs: a new parasitological processing technique, named TF-Test VetPet, and a microscopy image analysis pipeline based on deep learning methods. TF-Test VetPet improves image quality by reducing cluttering (i.e., eliminating artifacts), which favors automated image analysis. The proposed pipeline can identify three species of parasites in cats and five in dogs, distinguishing them from fecal impurities with an average accuracy of 98,6%. We also make available the two datasets with images of parasites of dogs and cats, which were obtained by processing fecal smears with temporary staining using TF-Test VetPet., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Giardia duodenalis is a flagellate protozoan that multiplies in the small intestine of a wide variety of hosts, animals and humans. It has a worldwide distribution, however it is considered a neglected disease by the World Health Organization (WHO). Nowadays, rabbits are being chosen as pets, especially by children. There are already reports of the occurrence of G. duodenalis in rabbits from other countries, but research has not been carried out in Brazil yet. Thus, the objective of our work was to verify the occurrence and molecularly characterize G. duodenalis that affect pet rabbits, through the polymerase chain reaction (PCR) in the northwest region of the state of São Paulo, Brazil. Fecal samples from 100 rabbits were collected, which later underwent a process of DNA extraction and amplification by nested-PCR (nPCR), using the SSU rRNA gene, and β-giardin (bg), glutamate dehydrogenase (gdh) and triosephosphate isomerase (tpi) to determine the assemblage. A questionnaire was answered by the owners with information about gender, age, deworming, diarrhea, water source, food, place of residence and contact with other animals. From those samples, 40 were positive for G. duodenalis. Good quality of the SSU rRNA gene by nPCR were obtained from two samples. For the first time, we report the occurrence of G. duodenalis assemblage A on pet rabbits in Brazil., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests, that research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Flying pigeons ( Columbia livia ) are extensively studied for their physical endurance and superior sense of orientation. The extreme physical endurance of which these birds are capable creates a unique opportunity to investigate the possible impact of long-distance flying on the taxonomy and metabolic function of the gut microbiota. This project was enabled by access to two groups of pigeons raised by the same breeder in the same conditions, except that one group was trained in long-distance flying and participated in multiple races covering a total distance of over 2600 km over a 9-week period. In contrast, the second group did not fly. The fecal microbiota was analyzed using 16S amplicon sequencing, and the taxonomy and metabolic function were inferred from this sequence data. Based on phylogenetic distance and metabolic function, flying and non-flying pigeons were found to harbor distinct bacterial microbiota. The microbiota taxonomy varied extensively between the birds, whereas the inferred metabolic potential was relatively stable. Age was not a significant determinant of the fecal microbiota profile. In flying birds, the metabolic pathways annotated with biosynthesis were enriched, representing 60% of the 20 metabolic pathways that were most closely associated with flying.

Cockatiels ( Nymphicus hollandicus ) are among the most commonly sold psittacines pets. The aim of this study was to evaluate the occurrence of Cryptosporidium spp. in domestic N. hollandicus and identify risk factors for this infection. We collected fecal samples from 100 domestic cockatiels in the city of Araçatuba, São Paulo, Brazil. Feces from birds of both genders and older than two months were collected. Owners were asked to complete a questionnaire to identify how they handle and care for their birds. Based on nested PCR targeting the 18S rRNA gene, the prevalence of Cryptosporidium spp. in the cockatiels sampled was 9.00%, 6.00% based on Malachite green staining, 5.00% based on modified Kinyoun straining, and 7.00% when the Malachite green was combined with Kinyoun. Applying multivariate logistic regression to test the association between Cryptosporidium proventriculi positivity and potential predictors showed that gastrointestinal alterations was a significant predictor ( p < 0.01). Amplicons from five samples were sequenced successfully and showed 100% similarity with C. proventriculi . In summary, this study demonstrates the occurrence of C. proventriculi in captive cockatiels.

Intestinal parasites inhabit the intestinal tract of humans and animals, causing damages whose severity depends on several factors related to the parasite and the host. Immunocompromised individuals are more likely to develop severe forms of parasitic infestation. The diagnosis of the gastrointestinal parasitosis is mainly performed by the examination of the feces, which consists of the direct visualization and identification of the parasites eliminated through the feces. These tests are generally low sensitive and the microscope slides contain a large number of impurities, which can impair the result of the diagnosis. In order to improve the diagnostic accuracy, a new parasitological technique called Three Fecal Test (TF‑Test) was developed. To further improve its diagnostic accuracy, few modifications of the original protocols have been made with the years. In this study the performance of these new techniques to detect gastrointestinal parasites in human and animal fecal samples was described and discussed in relation to the performance of other conventional coprological tests. It could be concluded that the TF‑Test conventional and modified can be used for the diagnosis of several human and animal parasites, with satisfactory results.

Objective: Ova and parasite (O&P) examination is recommended for the laboratory diagnosis of agents causing parasitic infections; however, this exam requires scientific and technological improvements to enhance its diagnostic validity. Dissolved air flotation (DAF) is an efficient technical principle separating suspended solids in a liquid medium. We aimed to develop and validate a new procedure for intestinal parasite detection with DAF., Methods: In this study, we collected samples from 500 volunteers, screened them by direct examination, and transferred the material to tubes using the Three Faecal Test (TF-Test) for triplicate DAF tests. We evaluated physical-chemical parameters and DAF prototype components through quantifying parasites recovered from floated and non-floated regions of the flotation column. The DAF operation protocol was validated with the gold standard results., Results: The 10% saturated volume proportion and cationic surfactant showed regularity and high parasite recovery (80%). Modifications of the needle device did not influence parasite recovery (p > 0.05). Sensitivity, specificity, accuracy and kappa agreement obtained with the DAF protocol were 91%, 100%, 93% and substantial (k = 0.64), respectively., Conclusion: The DAF principle could be used to process faecal samples in routine laboratory exams, enabling intestinal parasite detection., (© 2022 John Wiley & Sons Ltd.)

The Leishmania infantum (synonym, Leishmania chagasi) causes life-threatening infection, namely canine leishmaniosis (CanL), which is a chronic zoonosis prevalent in various countries and spread by the bite of the infected Lutzomyia female sandfly in South America. The objective of the study was to assess the effectiveness of a polymer matrix collar containing made up of 10% imidacloprid and 4.5% flumethrin for the prevention of canine leishmaniosis from the hyperendemic region falling under Araçatuba municipality (Brazil). The research included a total of 146 dogs chosen from 75 households. Test were initiated via physical examination; weighing and biological sample collection (blood, popliteal lymph node and conjunctival swab) of these dogs were done in March 2018 (Day 0; GA, control = 69, GB, treated = 77) to initiate laboratory tests. Post-inclusion, the animals were monitored on the 120th, 240th, 360th and 480th days, respectively. The usage of collars continued between 0 and 480 days before being substituted in second (D240) and fourth (D480) follow-up visits. On the whole, 25 dogs in GA (36.2%) and three in GB (3.9%) were found positive for L. infantum infection in a minimum of one diagnostic test used in the research. Therefore, the average collar effectiveness for protection from L. infantum infection was 89.2% (p < .01). In the last follow-up, the average incidence density rate for GA was 30.7%, whereas for GB, it was 2.9%. The imidacloprid/flumethrin collars evaluated in the research were found to be safe and extremely efficient for the prevention of L. infantum infection through Lutzomyia species among the large population of dogs in highly prone endemic regions. This is a dependable and efficient technique aimed at reducing the occurrence and propagation of this illness among the population of canines, which would eventually reduce the human-health-related hazards. In Brazil, Lutzomyia spp. is a leading vector of the infection; thus, the collar can be used to limit infection in dogs and humans., (© 2022 Wiley-VCH GmbH.)

Countless research is carried out until new discoveries are transformed into products or services available to the population. This trajectory can be slower and more costly or even impossible when irreproducible data are obtained in the most diverse fields of science. Thus, quality management appears as an essential tool to guarantee the reliability of academic research results. In this work, we demonstrate the applied strategy to implement a Quality Management System (QMS) in a research laboratory in Veterinary Parasitology and we highlight the adaptable quality requirements in this scientific research environment. For this, the Plan-Do-Check-Act (PDCA) quality tool was used, and two internal audits were performed, one before and one after implementation. The audits reached 67 (41.36%) and 157 (96.91%) points before and after implementation, respectively, with a significant difference between the moments studied. Thus, we demonstrate that the adoption of QMS principles in research is feasible. The methodology applied in this work can be adopted by managers from other laboratories interested in the implementation of quality standards as a support in the reproducibility of research., (Copyright © 2022 Elsevier Inc. All rights reserved.)

The aim of this study was evaluating the association and correlation between the diagnostics tests used for Leishmania spp. detection in dogs and ticks. We evaluated 99 dogs and 990 Rhipicephalus sanguineus. In dogs, we used bone marrow aspirates and lymph node fine-needle aspiration biopsy (FNAB) for direct parasitological examinations and real time-polymerase chain reaction (RT-PCR) and collected blood samples for enzyme-linked immunosorbent assays (ELISA). In ticks, two laboratory techniques [immunohistochemistry to lipophosphoglycan (IHC) and RT-PCR] were performed in the intestine, ovaries and salivary glands. With respect to the measurement of diagnostic performance in dogs, lymph node RT-PCR proved to be the best test followed by ELISA and bone marrow RT-PCR. In ticks, intestine IHC were considered as a gold standard for diagnosis of leishmaniasis with intestinal RT-PCR being the best diagnostic test. To arrive at the correlation between laboratory techniques for dogs and their ticks, we evaluated the diagnostic test used for dogs with tests performed in R. sanguineus, which used lymph node FNAB as the gold standard. The intestine IHC technique showed strongest association. We demonstrated that the best tissue for Leishmania spp. detection in dogs was the lymph node and the intestine in case of ticks. As for laboratory techniques, the isolated analysis of each species presented a strong agreement between immunohistochemistry and RT-PCR when compared to its gold standard. In addition, we concluded that the immunohistochemistry of ticks' intestines was a better technique for diagnosing Leishmania spp. in R. sanguineus, thereby showing almost perfect correlation with the lymph node FNAB., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

The parasitological examination of feces is recommended for the laboratory diagnosis of intestinal parasites due to its practicality, low-cost, and moderate diagnostic sensitivity. Dissolved Air Flotation (DAF) is an efficient technical principle used in other areas to separate dispersed solids. This study sought the preliminary evaluation of a proof-of-concept prototype as a tool for detecting species of parasites by adjusting DAF. Two DAF prototype units were developed to evaluate microbubbles' generation amidst fecal suspension and parasites' capture. For this evaluation, samples were screened and processed by the TF-Test technique (Control) and simultaneously by DAF device. The dimensional and attachment characteristics in the formation of the microbubbles were evaluated, and the percentage of parasitic recovery in floated and not-floated regions compared by Student's t-test. The second prototype unit proved to be more efficient in forming microbubbles with diameters between 34 and 170µm. The flotation tests showed a recovery of 73.27%, 58.12%, 37.85%, and 91.89% for Ascaris lumbricoides, Hymenolepis diminuta, Giardia duodenalis, and Strongyloides stercoralis, respectively. This study confirmed the selective interaction between microbubbles and parasite eggs and larvae during the flotation process using the DAF principle for the first time through imaging., (Copyright © 2021. Published by Elsevier B.V.)

The increasingly close proximity between people and animals is of great concern for public health, given the risk of exposure to infectious diseases transmitted through animals, which are carriers of more than 60 zoonotic agents. These diseases, which are included in the list of Neglected Tropical Diseases, cause losses in countries with tropical and subtropical climates, and in regions with temperate climates. Indeed, they affect more than a billion people around the world, a large proportion of which are infected by one or more parasitic helminths, causing annual losses of billions of dollars. Several studies are being conducted in search for differentiated, more sensitive diagnostics with fewer errors. These studies, which involve the automated examination of intestinal parasites, still face challenges that must be overcome in order to ensure the proper identification of parasites. This includes a protocol that allows for elimination of most of the debris in samples, satisfactory staining of parasite structures, and a robust image database. Our objective here is therefore to offer a critical description of the techniques currently in use for the automated diagnosis of intestinal parasites in fecal samples, as well as advances in these techniques., Competing Interests: The 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 Inácio, Gomes, Falcão, Martins dos Santos, Soares, Nery Loiola, Rosa, Nagase Suzuki and Bresciani.)

Toxoplasma gondii is a widespread protozoan parasite that causes infections in animals and humans. The presence of tissue cysts in cattle musculature evidences their epidemiological significance in transmission to humans. The present study aimed to evaluate the seroprevalence of and risk factors of T. gondii seropositivity in beef cattle in the municipality of Unaí, Minas Gerais state, Brazil. Bovine blood samples (n = 612) were collected from 31 herds. Risk factors associated with the animal-level and herd-level seroprevalence were analysed using a questionnaire that investigates the epidemiological aspects of toxoplasmosis. A search for anti-T. gondii antibodies was performed by means of an indirect fluorescent antibody test (IFAT). Analysis of 612 samples showed 369 animals positive for anti-T. gondii antibodies (IFAT ≥64), with a seroprevalence of 60.29% (95% confidence interval (CI): 56.35-64.10). The herd-level seroprevalence was 96.77% (95% CI: 79.00-99.58). Applying a multivariate logistic regression analysis, a test of the association between seroprevalence of T. gondii infection and the potential predictors showed that cats (domestic or wild) with access to the drinking water of cattle were a significant (P = 0.001) predictor. The seroprevalence of T. gondii in cattle destined for human consumption is widely distributed throughout the entire study area. Thus, integrated control measures and strategies are suggested to prevent T. gondii infection in cattle., (Copyright © 2021 Elsevier B.V. All rights reserved.)

This research had as objective to evaluate the occurrence and to characterize genetically the infections by Cryptosporidium in Mazama gouazoubira. By a non-invasive harvest methodology using trained sniffer dogs to locate fecal samples of cervids, 642 fecal samples were obtained from six Brazilian localities. The cervids species responsible for the excretion of each fecal sample were identified by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), using the mitochondrial cytochrome b target gene (cyst b) and the restriction enzymes Sspl, AflIII and BstN. From this identification, 437 fecal samples of M. gouazoubira were selected for research of Cryptosporidium spp. performed through negative staining with malachite green and polymerase chain reaction (nPCR), with the subunit of 18S rRNA gene, followed by sequencing the amplified products. In the samples that were diagnosed the presence of parasite species with zoonotic potential, genotyping was also performed using nPCR with the subunit of GP60 gene. Statistical analysis consisted of the Fisher exact test to verify the association of the presence of the enteroparasite in relation to the presence of cattle in each locality, and the McNemar tests and Kappa correlation coefficient used to compare the results obtained between the two diagnostic techniques. In the fecal samples of M. gouazoubira the occurrences of Cryptosporidium were diagnosed in 1.6% (7/437) and 1.1% (5/437), respectively, through nPCR and microscopy. Cryptosporidium. parvum was diagnosed in 100% (7/7) of the samples submitted to sequencing (18S gene). The IIaA16G3R1 subtype was diagnosed in five of the C. parvum samples submitted to genotyping (GP60 gene). This is the first world report of C. parvum in M. gouazoubira and subtype IIaA16G3R1 in cervids., (Copyright © 2020. Published by Elsevier B.V.)

Cryptosporidium spp. and Giardia duodenalis infect the gastrointestinal tracts of animals and humans. Both parasite groups are distributed worldwide and cause significant economic losses in animal productivity. Infected hosts presenting with and without clinical manifestations can eliminate infective forms of these protozoa, which are particularly important to One Health. Compared to the published research on cattle, relatively few studies have examined the epidemiology of cryptosporidiosis and giardiasis in buffaloes. This short review describes the global occurrence of Cryptosporidium spp. and G. duodenalis in buffaloes, including the molecular techniques employed for the identification of species/assemblages and genotypes of these protozoa. Genetic analyses of isolates of G. duodenalis and Cryptosporidium spp. from various sources (environmental, animal, and human) have been performed to investigate their epidemiology. In buffaloes, the species Cryptosporidium parvum, Cryptosporidium ryanae, Cryptosporidium bovis , and Cryptosporidium suis -like have been characterized, as well as assemblages A and E of G. duodenalis . We demonstrate that buffaloes can be infected by species of Cryptosporidium spp. and G. duodenalis assemblages with zoonotic potential. Epidemiological studies that utilize molecular biology techniques represent an important resource for efforts to control and prevent the spread of these protozoans., (Copyright © 2020 de Aquino, Inácio, Rodrigues, de Barros, Garcia, Headley, Gomes and Bresciani.)

Background: Prenatal toxoplasmosis remains a neglected disease worldwide and few government programs focusing on its prevention are available. Success in these programs has been extensively reported in the literature, yet the strategies used for their implementation, as a model for such actions in different communities, have not been described., Objective: To describe the aspects of prenatal care strategies in 13 municipalities within the regional healthcare unit of Araçatuba, in the northwestern region of the state of São Paulo in 2017, focusing on congenital toxoplasmosis., Design and Setting: Descriptive study on prenatal healthcare within the Brazilian National Health System, in 13 participating municipalities., Methods: Data on serological tests, referral clinics, notifications, healthcare strategies, health education and drugs for infected children were requested through a questionnaire that was sent via e-mail to people responsible for healthcare services in these municipalities., Results: Major differences relating to diagnoses, reference outpatient clinics, notifications, health education and healthcare and drugs for infected children were reported among the prenatal strategies of these 13 municipal healthcare services., Conclusion: The lack of standardized prenatal strategy in the study area may compromise the prevention of infection. However, our identification of each aspect of prenatal care corroborates the need to implement a healthcare surveillance program relating to congenital toxoplasmosis.

Toxoplasmosis and neosporosis are diseases with worldwide distribution that are associated with reproductive problems in livestock and responsible for economic losses. This review presents an overview of the current knowledge relative to these diseases in water buffalo ( Bubalus bubalis ). In general, buffalo are considered resistant to clinical toxoplasmosis because there are studies only reporting serological evidence of natural infection in these animals. Studies have described age, poor hygienic status of the farm, and presence of cats as risk factors for the development of Toxoplasma gondii infection in buffalo. It must be highlighted that buffalo meat, which does not receive adequate freezing treatment, could be a potential source for toxoplasmic human infection as well as the importance of raw buffalo milk in the transmission of toxoplasmosis to human beings. Neospora caninum is considered one of the major causes of abortion and responsible for huge economic losses in cattle. Vertical transmission is the main route to infect calves, and is responsible for maintaining the parasite within a herd. In buffalo, vertical transmission is also described; moreover, although there are indications that N. caninum may be associated with abortion in dairy buffalo, the reproductive importance of neosporosis is apparently lower in buffalo relative to cattle. Most studies have identified a higher time of exposition to N. caninum oocysts relative to age. The household system was also described as a risk factor for infection, possibly due to persistent contact between the home-raised buffalo and canids. The fetal immune competence of buffalo is similar to bovine, and buffalo fetus are highly susceptible to infection during the first trimester of pregnancy, indicating that N. caninum may be an abortigenic agent in buffaloes. Alternatively, it is interesting to note there is evidence that the inflammatory response in pregnant buffalo infected with N. caninum is mild enough to avoid abortion in most cases. It is proposed that the possible transmission of toxoplasmosis through unprocessed milk and buffalo meat may occur, which is important in terms of public health. Additionally, there is strong evidence to suggest that N. caninum may be associated with abortion in buffalo., (Copyright © 2020 de Barros, Garcia, Bresciani, Cardim, Storte and Headley.)

In this study, we evaluated the performance of a new enzyme-linked immunosorbent assay (ELISA) variant known as indirect "plasmonic ELISA" (pELISA) for the detection of Leishmania spp. infection. Serum samples from 170 dogs from an area where canine leishmaniosis (CanL) is endemic and from 26 healthy dogs from a nonendemic area were tested by indirect pELISA, and the results were compared to those of an indirect ELISA (both with recombinant antigen rK28) and those of an immunochromatographic test (dual-path platform, TR-DPP®) using real-time PCR on blood samples or conjunctival swabs as the gold standard. The pELISA, indirect rK28 ELISA and the TR-DPP® immunochromatographic test presented sensitivities of 94.7%, 89.5% and 79.0% and specificities of 100%, 92.7% and 91.5%, respectively. The analysis of the results revealed that the specificity of the indirect pELISA was greater than that of the method recommended by the Ministry of Health in Brazil and may increase the feasibility of diagnosis in resource-constrained countries because it does not require sophisticated instruments to read. Thus, this method can be used as an additional tool for the detection of Leishmania spp. infection in these areas., (© 2019 John Wiley & Sons Ltd.)

Since the early 20th century, the detection of intestinal parasites has improved with the development of several techniques for parasitic structures recovery and identification, which differ in sensitivity, specificity, practicality, cost, and infrastructure demand. This study aims to review, in chronological order, the stool examination techniques and discuss their advantages, limitations, and perspectives, and to provide professionals and specialists in this field with data that lays a foundation for critical analysis on the use of such procedures. The concentration procedures that constitute the main techniques applied in routine research and in parasitological kits are a) spontaneous sedimentation; b) centrifugation-sedimentation with formalin-ethyl acetate; and c) flotation with zinc sulfate solution. While selecting a technique, one should consider the purpose of its application and the technical-operational, biological, and physicochemical factors inherent in the procedures used in stool processing, which may restrict its use. These intrinsic limitations may have undergone procedural changes driven by scientific and technological development and by development of alternative methods, which now contribute to the improvement of diagnostic accuracy.

Introduction: Canine visceral leishmaniasis (CVL) is a public health problem, and its prevalence is associated with the coexistence of vectors and reservoirs. CVL is a protozoonosis caused by Leishmania infantum that is endemic in the southeast region of Brazil. Thus, vector and canine reservoir control strategies are needed to reduce its burden. This study aimed to verify the CVL seroprevalence and epidemiology in a municipality in Southeast Brazil to initiate disease control strategies., Methods: A total of 833 dogs were subjected to Dual Path Platform (DPP) testing and enzyme-linked immunosorbent assays. For seropositive dogs, epidemiological aspects were investigated using a questionnaire and a global position system. The data were submitted to simple logistic regression, kernel estimation, and Bernoulli spatial scan statistical analysis., Results: The overall CVL-confirmed seroprevalence was 16.08%. The 28.93% in the DPP screening test was associated with dogs maintained in backyards with trees, shade, animal and/or bird feces, and contact with other dogs and cats, with sick dogs showing the highest chances of infection (odds ratio, 2.6; 95% confidence interval, 2.38-1.98), especially in residences with elderly people. A spatial analysis identified two hotspot regions and detected two clusters in the study area., Conclusions: Our results demonstrated that residences with elderly people and the presence of trees, shade, feces, and pet dogs and cats increased an individual's risk of developing CVL. The major regions where preventive strategies for leishmaniasis were to be initiated in the endemic area were identified in two clusters.

Objectives: Human enteroparasites are considered a serious public health problem in underdeveloped countries located in world regions with tropical, subtropical and equatorial climates. These parasites are commonly diagnosed by the Parasitological Examination of Faeces (PEF), performed by conventional techniques and/or commercial kits that result in tests with low-to-moderate sensitivity, due to the use of destructive chemical solvents to parasite structures, and to present excess adipose substance and digestive residues in their microscopic slides. In order to improve the efficacy of these tests/examinations, this study aimed to investigate a new alternative for the PEF, with the use of Aqueous Biphasic System (ABS)., Methods: For this, four ABSs containing poly (ethylene glycol), PEG (PEG-4000 and PEG-6000), dipotassium phosphate and sodium citrate at different concentrations in the biphasic systems were evaluated with faecal samples containing eggs of Ascaris lumbricoides., Results: The ABS consisting of PEG-4000 and dipotassium phosphate, at concentrations of 55% w/w and 20% w/w, respectively, achieved 100% satisfactory results compared to the conventional TF-Test technique in terms of preservation and concentration of A. lumbricoides eggs, with adequate separation of digestive residues, without using a centrifuge or chemical solvents that may cause harm to the parasites., Conclusions: This study presents ABS as a new low-cost technical principle for the detection of parasite eggs in PEF. The new technique is simple, fast, non-toxic, not harmful to the parasite and does not require a centrifuge., (© 2019 John Wiley & Sons Ltd.)

Based on our initial observations showing that mice consuming a probiotic product develop more severe cryptosporidiosis, we investigated the impact of other dietary interventions on the intracellular proliferation of Cryptosporidium parvum and C. tyzzeri in the mouse. Mice were orally infected with oocysts and parasite multiplication measured by quantifying fecal oocyst output. High-throughput sequencing of 16S ribosomal RNA amplicons was used to correlate oocyst output with diet and with the composition of the intestinal microbiota. On average, mice fed a diet without fiber (cellulose, pectin and inulin) developed more severe infections. As expected, a diet without fibers also significantly altered the fecal microbiota. Consistent with these observations, mice fed a prebiotic product sold for human consumption excreted significantly fewer oocysts. The fecal microbiota of mice consuming no plant polysaccharides was characterized by a lower relative abundance of Bacteroidetes bacteria. Since bacterial metabolites play an important role in the physiology of intestinal enterocytes, we hypothesize based on these observations that the impact of diet on parasite proliferation is mediated primarily by the metabolic activity of the anaerobic microbiota, specifically by the effect of certain metabolites on the host. This model is consistent with the metabolic dependence of intracellular stages of the parasite on the host cell. These observations underscore the potential of dietary interventions to alleviate the impact of cryptosporidiosis, particularly in infants at risk of recurrent enteric infections., Competing Interests: The authors have declared that no competing interests exist.

We performed molecular characterization of Giardia duodenalis in buffalo calves from the Southwest region of São Paulo State, Brazil. A total of 183 fecal samples of Murrah breed buffaloes up to six months of age were collected. We examined these samples by the polymerase chain reaction (PCR) targeting the small-subunit ribosomal RNA gene and positive samples were characterized using additional PCR assays targeting a portion of the beta-giardin, the glutamate dehydrogenase and the triose-phosphate isomerase genes. Based on the SSU rRNA nPCR, the presence of G. duodenalis was confirmed in 12 (6.56%) of fecal samples, of these, five, four and three samples were positive for the tpi , bg and gdh genes, respectively. Assemblage identification by sequencing was successful in 6 of 12 samples and sequence analysis showed 100% genetic similarity with G. duodenalis assemblage E. This observation represents the first detection of G. duodenalis assemblage E in buffaloes calves in Brazil., Competing Interests: The authors declare no conflict of interest., (© 2019 Published by Elsevier Inc. on behalf of International Association of Food and Waterborne Parasitology.)

Human cryptosporidiosis is caused primarily by two species of apicomplexan parasites, Cryptosporidium parvum and C. hominis. Although infection of cell monolayers with sporozoites does not support the complete parasite life cycle, the in vitro system is used to study the asexual phase of multiplication, which consists of two generations of merogony. To better understand host-parasite interaction and to gain insight into gene regulatory processes driving the complex life cycle of Cryptosporidium parasites, we analyzed the transcriptome of C. parvum in oocysts, sporozoites and infected cell monolayers 2-48 h post-infection. Analysis of RNA-Seq data from replicate oocyst, sporozoite and intracellular samples revealed significant differences between transcriptomes expressed outside and inside the host cell. Compared to the transcriptome found in the host cell, the oocyst transcriptome is less diverse. Biological processes significantly over-represented intracellularly relate to biosynthetic processes. Genes significantly overexpressed in oocysts show evidence of specialized functions not found in other Apicomplexa. A more comprehensive view of gene regulation during the Cryptosporidium life cycle will require the analysis of later time points during the infection, particularly of the poorly studied sexual phase of the life cycle.

Objectives: Laboratory diagnosis of Schistosoma mansoni eggs is routinely performed by conventional quantitative techniques through the parasitological examination of human faeces. However, the diagnostic sensitivity of this type of exam varies from low to moderate. We aimed to develop a new parasitological technique called TF-Test Quantified (TFT-Quant), for the quantitative detection of S. mansoni eggs in human faeces., Methods: Four study stages were performed using 43 mice infected by S. mansoni cercariae. These experiments allowed the definition of an operational protocol for TFT-Quant and a comparison of this new technique to the conventional Helm-Teste technique., Results: The results show a good diagnostic efficacy of TFT-Quant, which reached 100% in sensitivity and specificity, indicating an 'Almost Perfect' Kappa (k) agreement., Conclusions: This new technique provided a quantitative gain in the detection of S. mansoni eggs, largely free of faecal debris. The next stage of this study is the field validation of the TFT-Quant technique with the use of human faecal samples from an endemic region for mansonic schistosomiasis in Brazil (Microregion Jequitinhonha, State of Minas Gerais). In parallel to this validation, computational algorithms will be developed to allow the automated quantitative diagnosis of S. mansoni eggs., (© 2019 John Wiley & Sons Ltd.)

Canine monocytic ehrlichiosis (CME) is a disease caused by the obligate intracellular bacterium Ehrlichia canis. Tropical lineages of Rhipicephalus sanguineus ticks play an essential role in the transmission of this pathogen. The aim of the present study was to evaluate the prevalence of E. canis DNA in tissue from R. sanguineus ticks in areas endemic for CME in Brazil and quantify levels of E. canis DNA in dissected tissues from these samples. A total of 720 ticks were collected from 72 dogs (36 dogs from the city Araçatuba in São Paulo state and 36 from Campo Grande in the state of Mato Grosso do Sul). Ticks were dissected to collect the guts, ovaries and salivary gland. A quantitative polymerase chain reaction (qPCR) targeting the disulphide bond formation (dsb) protein gene was performed to quantify the level of E. canis infection. The E. canis dsb-qPCR assay was positive for 31.9, 10, and 15.2% of the gut, ovary, and salivary glands, respectively. The average gut, ovary, and salivary gland bacterial load estimated by qPCR was 1.21 × 103, 2.60 × 103, and 4.92 × 103 gene copies/µl, respectively. This is the first report of E. canis DNA in ovaries of R. sanguineus ticks parasitizing dogs in these CME-endemic areas. These observations raise the possibility of E. canis trans-ovarial transmission., (© The Author(s) 2018. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

The aim of this study was to evaluate the prevalence of and diagnostic methods for Cryptosporidium spp. in caged adult exotic parrots from Southern and Southeastern Brazil. Oocysts were purified from fecal samples from 463 psittacines by centrifugal-flotation in Sheather's sugar solution. Cryptosporidium spp. were detected by malachite green negative staining and nested PCR targeting the 18S rRNA gene. Cryptosporidium species were identified by sequencing nested PCR amplicons. Samples were also tested by duplex real-time PCR targeting the 18S rRNA gene of Cryptosporidium galli and Cryptosporidium avian genotype III. The prevalence rates of Cryptosporidium spp. determined by microscopy and nested PCR were 3.0% (14/463) and 5.0% (23/463), respectively. The nested PCR/sequencing identified avian genotype III (1.7%; 8/463), Cryptosporidium parvum (0.9%; 4/463) and Cryptosporidium canis (0.2%; 1/463). Duplex real-time PCR was positive for gastric Cryptosporidium in 9.5% (44/463) of the samples. Among them, 1.9% (9/463) were positive for C. galli, 5.8% (27/463) were positive for avian genotype III and 1.7% (8/463) showed mixed infections with C. galli and avian genotype III. With regards to the positive detection of Cryptosporidium spp., there was no statistically significant difference between nested PCR and microscopic analysis (p = .1237), and a fair agreement existed between them (Kappa = 0.242). A statistically significant difference (p < .0001) and fair agreement (Kappa = 0.317) were obtained between nested PCR/sequencing and duplex real-time PCR for the detection of gastric Cryptosporidium. We determined that nested PCR and duplex real-time PCR are the best options for the detection of Cryptosporidium spp. and gastric Cryptosporidium, respectively, and that avian genotype III is the most common Cryptosporidium genotype/species in psittacines., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Despite of the role of domestic dogs as reservoirs for threatening viral diseases for wild carnivores, few studies have focused to identify circulation of viruses among dogs living in human/wildlife interfaces. To identify canine parvovirus (CPV) types circulating in dogs living in an Atlantic forest biome, faecal samples (n = 100) were collected at the same period (one week) corresponding to each of four areas, during 2014 to 2016 and corresponded to 100 different individuals. CPV was isolated in cell culture from 67 out 100 (67%) samples from healthy dogs. Cytopathic effects were characterized by total or partial cell culture lysis. Genome sequences of CPV-2a (10%), CPV-2b (7%) and CPV-2c (50%) were concomitantly detected by PCR and nucleotide sequencing. The current study addresses the importance of monitoring CPV circulation among dogs presenting potential contact with wildlife species.

The carrier pigeon and the domestic pigeon are different breeds of the species Columba livia. Carrier pigeons are used for recreational activities such as bird contests and exhibitions. Due to the close contact with humans, these birds may potentially represent a public health risk, since they can host and disseminate zoonotic parasites, such as those belonging to the genus Cryptosporidium (phylum Apicomplexa). The purpose of this work was the detection by microscopic and molecular techniques of Cryptosporidium spp. oocysts in fecal samples of carrier pigeons, and subsequently to sequence the 18S ribosomal RNA marker of positive samples to identify the species. A total of 100 fecal samples were collected individually in two pigeon breeding facilities from Formiga and Araçatuba, cities located in Minas Gerais state and São Paulo state, Brazil, respectively. The age of the birds ranged from one to 12 years; 56 were females and 44 males. Fecal smears were stained with negative malachite green, whereas the molecular characterization was based on the sequence of a ∼800bp fragment of the 18S rRNA gene. Microscopic examination of fecal smears revealed 4% (4/100) oocyst positivity. On the other hand, 7% (7/100) of positivity were found using nested PCR. Three samples were 99% to 100% similar to Cryptosporidium parvum 18S rDNA type A (Genbank AH006572) and the other three samples had 99% to 100% similarity to C. parvum 18S rDNA type B (Genbank AF308600). To our knowledge, this is the first report of C. parvum oocysts in carrier pigeons., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Mosquitoes can act as vectors of important diseases such as malaria, dengue, Zika virus, yellow fever, Chikungunya and Mayaro fever, in addition to filariasis. The use of insecticides, larvicides, bed nets and repellents, besides the use of drugs as chemoprevention and the treatment of the sick are currently the pillars of the control of these vectors. We studied the biological control against of Anopheles darlingi , Aedes aegypti and Culex quinquefasciatus larvae using shrimps of the species M. pantanalense , M. amazonicum , M. brasiliense and M. jelskii . Larvae of mosquitoes were collected from the breeding environment and placed in a 500 and 1000 l tank containing 60 shrimps/m 2 . The predatory activity was evaluated for 30 days and, in all groups it was observed that 100% of the larvae were consumed in few minutes. In the environment, these same species of crustaceans were released in water bodies with the presence of larvae of these insects. In just 72 h there was a marked reduction of the larvae in the release sites of shrimps. Similarly, there was a reduction in the number of adult mosquitoes caught near the breeding sites, allowing to infer that, in places where the crustaceans were released, the predatory activity on the larvae of mosquitoes was sufficient to reduce the number of adult mosquitoes p ≤ 0,05. This is the first description of the predatory activity of M. pantanalense , M. amazonicum , M. brasiliense and M. jelskii on An. darlingi , A. aegypti and C. quinquefasciatus larvae, constituting an important tool of biological control of these parasites-vectors.

This study aimed to investigate the presence of Toxoplasma gondii in semen, testicle and epididymis tissues of cats experimentally infected by this coccidium. A total of 12 male felines without a definite breed that were of reproductive age and serologically negative for T. gondii were selected and distributed to the following three experimental groups: GI, inoculated with 600 tissue cysts of the P strain of T. gondii (isolate III); GII, inoculated with 2×10 5 tachyzoites of the RH strain (isolate I); and GIII, not inoculated (control group). Prior to inoculation (day -7 and 0) and on post inoculation days (PIDs) 7, 14, 21, 28, 42, 56, and 70, all felines were subjected to assessments of anti-T. gondii IgG by indirect immunofluorescence (IIF) and assessments of parasitemia. Collection of semen (electroejaculation) was performed on the specified dates, followed by nested PCR and bioassays in mice to detect T. gondii. On PID 70, all 12 felines were orchiectomized, and the presence of the parasite in the testicles and epididymides was evaluated by nested PCR, murine bioassay, and histopathological and immunohistochemical analyses. All felines inoculated with T. gondii (GI and GII) seroconverted to the toxoplasmic infection after PID 14; on PID 7, seroconversion of three felines (P4, RH2 and RH4) could observed, and all exhibited detectable titers by PID 64. The GII felines exhibited greater serological titers compared with GI felines. The maximum serological titer (IgG) was observed in feline RH3 (titer 1024), while in other experimental felines, a maximum titer of 256 was detected. Parasitemic peaks were diagnosed in all felines of groups I and II from PIDs 7-42. A total of five parasitemic peaks were diagnosed in GI and nine in GII. In none of the experimental time points was the presence of T. gondii diagnosed in seminal samples collected from the felines or in the testicle or epididymis tissues collected from these animals. Thus, sexual transmission in domestic cats does not appear to be a major route of T. gondii infection, possibly demonstrating the tendency of this protozoan to develop a response directed to the formation and excretion of oocysts in the feces of these definite hosts, which act as its main route of perpetuation in the environment., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Due to its important zoonotic potential, cryptosporidiosis arouses strong interest in the scientific community, because, it was initially considered a rare and opportunistic disease. The parasitological diagnosis of the causative agent of this disease, the protozoan Cryptosporidium spp., requires the use of specific techniques of concentration and permanent staining, which are laborious and costly, and are difficult to use in routine laboratory tests. In view of the above, we conducted the feasibility, development, evaluation and intralaboratory validation of a new parasitological technique for analysis in optical microscopy of Cryptosporidium spp. oocysts, called TF-Test Coccidia, using fecal samples from calves from the city of Araçatuba, São Paulo. To confirm the aforementioned parasite and prove the diagnostic efficiency of the new technique, we used two established methodologies in the scientific literature: parasite concentration by centrifugal sedimentation and negative staining with malachite green (CSN-Malachite) and Nested-PCR. We observed good effectiveness of the TF-Test Coccidia technique, being statistically equivalent to CSN-Malachite. Thus, we verified the effectiveness of the TF-Test Coccidia parasitological technique for the detection of Cryptosporidium spp. oocysts and observed good concentration and morphology of the parasite, with a low amount of debris in the fecal smear., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The prevalence study of Leishmania spp. in hematophagous insects captured from the environment in bat roosts and pigeon nests, or feeding their hosts (cattle, pigs, horses, dogs and humans) in urban, peri-urban and rural areas, between 2012 and 2014. For this study, the amastigotes present in these insects were detected by histochemical and PCR techniques. Positive gene amplification for Leishmania was found in two horseflies of the species Tabanus importunus collected in the environment, and amastigote forms of Leishmania spp., as well as erythrocytes and leukocytes, were histochemically detected in one of that insect. The other analyzed insects were not positive by PCR our by direct parasitological examination. Only horseflies captured in urban and peri-urban areas were positive. During the collection, no phlebotomine sand flies were captured in rural areas far from the city limits. It can be concluded that the discovery of horseflies positive for Leishmania spp. in urban and peri-urban areas indicates the likelihood that urban areas and their surroundings provide vector parasites with an environment suitable for the spread and consequent perpetuation of the biological cycle of this protozoan.