Infecção concomitante experimental de Rattus norvegicus por Toxocara canis e Toxoplasma gondii: estudo comportamental e histopatológico

A hipótese de \"manipulação comportamental\" supõe que um parasito pode alterar o comportamento de seu hospedeiro visando aumentar a probabilidade de completar seu ciclo evolutivo. Tais alterações aumentariam a taxa de transmissão hospedeirohospedeiro, assegurando ao parasito ou a seus propágulos o encontro de novo hospedeiro. A possibilidade de infecções parasitárias provocarem mudanças comportamentais em seus hospedeiros e a elevada frequência com que o acometimento de seres humanos por larvas de Toxocara e cistos de Toxoplasma ocorre, têm chamado à atenção de pesquisadores interessados no estudo das relações hospedeiro-parasita. Na infecção por Toxoplasma gondii e Toxocara canis, cistos e larvas estão presentes em diversos locais anatômicos incluindo musculatura, coração, pulmões, olhos e cérebro. A presença de parasitos no cérebro dá oportunidade de manipulação do comportamento do hospedeiro. Entretanto, não se sabe qual ou quais mecanismos estão envolvidos no processo de manipulação do comportamento. Os objetivos do presente estudo foram verificar alterações na ansiedade, medo, memória e aprendizagem de Rattus norvegicus experimentalmente infectados por Toxocara canis e/ou Toxoplasma gondii em dois períodos após infecção, bem como a localização das larvas e cistos e presença de placas beta amiloide ( A) na região do hipocampo no tecido cerebral desses roedores corado pela técnica de Hematoxilina e Eosina (HE), e Vermelho de Congo, respectivamente. Foram utilizadas 40 exemplares fêmeas da espécie Rattus norvegicus, com seis a oito semanas. Os animais foram divididos em quatro grupos: Toxocara - 10 ratos infectados com 300 ovos de Toxocara canis, Toxoplasma -10 ratos infectados com 10 cistos de Toxoplasma gondii, Infecção dupla - 10 ratos infectados com 300 ovos de Toxocara canis e 10 cistos de Toxoplasma gondii, e controle - 10 ratos sem infecção. Nos dias 40, 41, 70 e 71 após a infecção, os animais dos grupos infectados e controle foram submetidos à avaliação no Labirinto em Cruz Elevado e Campo aberto. Aos 120 após infecção foi feita avaliação da memória, aprendizado e aversão a urina de gato dos animais no Labirinto de Barnes. No final das análises comportamentais os animais foram levados a eutanásia para retirada do cérebro e confecção dos cortes histológicos preparados em HE e Vermelho de Congo. Os resultados mostraram efeito ansiolítico para ambas as infecções, principalmente para Toxoplasma gondii. Não houve comprometimento da memória e aprendizado no LB, porém os animais infectados por Toxocara canis ou Toxoplasma gondii apresentaram menor tempo para encontrar a toca com urina e entrar nela. A leitura dos cortes histológicos corados com HE mostraram larvas de Toxocara canis e cistos de Toxoplasma gondii em regiões do sistema nervoso central dos animais relacionadas com memória e aprendizado. As lâminas coradas com Vermelho de Congo apresentaram placas beta amiloides ( A) em metade dos animais infectados por Toxoplasma gondii. Conclui-se que a infecção por ambos os parasitos apresenta efeito ansiolítico quando ocorre infecção única. Quando a Infecção ocorre concomitantemente há modulação no comportamento. Além disso, ratas infectadas com infecção única apresentam-se menos aversivas à urina de gatos.

Heterogeneity in testing practices for infections during pregnancy: national survey across Switzerland.

QUESTION Detection and treatment of infections during pregnancy are important for both maternal and child health. The objective of this study was to describe testing practices and adherence to current national guidelines in Switzerland. METHODS We invited all registered practicing obstetricians and gynaecologists in Switzerland to complete an anonymous web-based questionnaire about strategies for testing for 14 infections during pregnancy. We conducted a descriptive analysis according to demographic characteristics. RESULTS Of 1138 invited clinicians, 537 (47.2%) responded and 520 (45.6%) were eligible as they are currently caring for pregnant women. Nearly all eligible respondents tested all pregnant women for group B streptococcus (98.0%), hepatitis B virus (HBV) (96.5%) and human immunodeficiency virus (HIV) (94.7%), in accordance with national guidelines. Although testing for toxoplasmosis is not recommended, 24.1% of respondents tested all women and 32.9% tested at the request of the patient. Hospital doctors were more likely not to test for toxoplasmosis than doctors working in private practice (odds ratio [OR] 2.52, 95% confidence interval [CI] 1.04-6.13, p = 0.04). Only 80.4% of respondents tested all women for syphilis. There were regional differences in testing for some infections. The proportion of clinicians testing all women for HIV, HBV and syphilis was lower in Eastern Switzerland and the Zurich region (69.4% and 61.2%, respectively) than in other regions (range 77.1-88.1%, p <0.001). Most respondents (74.5%) said they would appreciate national guidelines about testing for infections during pregnancy. CONCLUSIONS Testing practices for infections in pregnant women vary widely in Switzerland. More extensive national guidelines could improve consistency of testing practices.

Análise do perfil de encaminhamento da atenção primária para pré-natal de alto risco: desafios para o ensino de competências em saúde materno-infantil

OBJECTIVE: to identify a profile of the main causes of inappropriate referrals from primary care to specialized services, as strategy for the curriculum development of core competencies related to maternal health. METHODS: a cross-sectional study was performed using document analysis of all referrals of pregnant women from primary care to the high-risk pregnancy service, state of Rio Grande do Norte, Brazil. All pregnant women referred from June to December 2014 (n = 771) were included. According to their causes the referrals were categorized as adequate, inadequate or inconclusive. RESULTS: a total of 188 referrals were classified as inadequate (24.4%) and 93 inconclusive (12.1%) totalizing 36.5% of inappropriate referrals. The main causes identified in these inappropriate referrals were: low-risk pregnancy (12.8%), unconfirmed hypertension (12.1%), risk of abortion (8.9%), teenage pregnancy (7.1%) , toxoplasmosis (5.3%), Rh incompatibility (4.6%) and urinary tract infection (4.3%). These data contributed to the formulation of the following products: 1) a continuing education program for health professionals working in primary care, undergraduate students and residents; and 2) development of a virtual platform to support professionals who need to refer patients to high-risk pregnancy service. CONCLUSION: the results of this study are relevant in the current context of education of health professionals, with potential for positively impact not only in the development of skills related to maternal health in undergraduate and graduate education, as well as contributing for improvement of the health care of the population.

Parasitos de importância para a caprinocultura e a ovinocultura potiguar

The goat and sheep industry shows up as an agricultural activity of great importance for the semiarid Northeast. However, the sheep and goats production is made with various difficulties. Among them, parasitic infections, particularly helminth infections of the gastrointestinal tract, the eimeriosis and toxoplasmosis; this one related to problems in reproduction. For this reason, the aim of this study is to to make a survey of the occurrence and some determinants of parasitic diseases that affect small ruminant flocks of the microregions Natal, Macaíba, Litoral Sul, Angicos, Vale do Açu and Borborema Potiguar. Thereunto, epidemiological tools were applied with producers, keepers or guardians of herds and also held collections of blood and feces of animals in eight properties located in seven municipalities of these microregions. The parasite load of the animals was determined through eggs and oocysts counting per gram of feces EPG and OPG, respectively. In addition, the recovery of infective larvae was made. Blood samples were used to measure the globular cell volume and the search for anti-Toxoplasma gondii IgG in sheep serum, by Enzyme Linked Immune Sorbent Assay (ELISA). For categorical variables, the statistical analysis was performed using Poisson regression, with significance level of 0.05. The analysis of the instruments showed that ivermectin is the anthelmintic used in 85,71% of properties. From the total of feces samples of the sheep (n = 179), 53,07% were positive for helminth eggs and 48,04% were positive for oocysts of Eimeria. From the samples of faeces of goats (n = 133), 72,18% were positive for helminth eggs and 96,99% for oocysts of Eimeria. The lowest EPG and OPG count was observed in the micro region of Angicos. Most of the EPG count was found in the micro region Litoral Sul and the OPG count in the micro-region Borborema Potiguar. Both cases the difference was statistically significant(p- value0,000)The most prevalent helminth genus found was Haemonchus, present in 49,87% of the sheep and 80,42% of goats. The average of hematocrit ranged from 22,91 to 33,25 in sheep and from 22,62 to 28,25 in goats. The prevalence of anti-Toxoplasma gondii IgG ranged from 63,33% to 100,00%. The goats showed to be more susceptible to infections by parasites of the gastrointestinal tract than the sheep. In all the properties was observed high prevalence of infection by T. gondii, with the lowest percentages recorded in the micro regions Angicos and Borborema Potiguar.

Utilização de peptídeos sintéticos derivados de moléculas imunodominantes de Toxoplasma gondii para diagnóstico sorológico da toxoplasmose em ovinos e galinhas caipiras

Chapter I - The obligate intracellular parasite Toxoplasma gondii is the causative agent of toxoplasmosis, a disease that affects humans and generates economic losses in farm animals. When prevention fails disease refers to the diagnosis and subsequent treatment if the individual is diagnosed as positive. Therefore, the development of new accurate diagnostic tools for detecting T. gondii infection is a need in particular to determine the environmental source of infection which can result in more appropriate public health policies against different routes of infection and prevent potential damage that toxoplasmosis can cause when animals are infected. Chapter II - The domestic chicken (Gallus gallus domesticus) are considered epidemiological sentinels, still representing a major source of recombinant strains when predated by cats, it is common to find them with multiple infections. We evaluate the diagnostic potential of six synthetic peptides (SAG2Y, MIC1, M2AP, GRA10, ROP2 and ROP7) predicted in silico from tachyzoites immunodominant markers of T. gondii in samples from naturally infected chickens, comparing synthetic peptides with antigen total soluble (STAg). In general, our results demonstrated that reactivity rates and positivity for these peptides are similar to the STAg, and the ROP7 peptide and the combination of peptides MIC1+ROP2 have significant sensitivity, confirming them as potential diagnostic tools for the diagnosis of toxoplasmosis in chickens. Chapter III - Sheep (Ovis aries) are commonly infected with Toxoplasma gondii due to his eating habits. Infection in pregnant sheep can have serious consequences such as embryonic death, fetal resorption, mummification, and neonatal death. One concern regarding the infection in these animals is that the meat can be a source of contamination to humans and other carnivores. Therefore perform accurate diagnosis in these animals is of fundamental importance. In the present study we evaluated the potential of new synthetic peptides as a diagnostic tool. Synthetic peptides (SAG2Y, SRS52A, MIC14, GRA4, GRA10 and GRA15) were predicted in silico from immunodominant proteins of T. gondii. We determine the levels of IgG antibodies using sera obtained from two farms in the city of Uberlândia. Analyzing the results together, the peptide combination GRA10+GRA15 (Accuracy = 0,82) showed better characteristics compared with the other mixtures. This preparation could be better analyzed with an antigenic mixture potential use in the diagnosis of toxoplasmosis in sheep and other species.

Enrofloxacina e toltrazuril reduzem a infecção por Toxoplasma gondii em células trofoblásticas humanas e em explantes de vilos placentários humanos de terceiro trimestre

The classical treatment for congenital toxoplasmosis is based on combination of sulfadiazine and pyrimethamine plus folinic acid. Due to teratogenic effects and bone marrow suppression caused by pyrimethamine, the establishment of new therapeutic targets is indispensable to minimize the undesirable effects and improve the control of the infection. Previous studies demonstrated that enrofloxacin and toltrazuril were able to control the infection triggered by Neospora caninum and Toxoplasma gondii. Therefore, the aim of this present study was evaluate the efficacy of enrofloxacin and toltrazuril in the control of T. gondii proliferation in human trophoblast cells (BeWo lineage) and in human villous explants from third trimester. BeWo cells and villous were treated with several concentrations of enrofloxacin, toltrazuril, sulfadiazine, pyrimethamine or association (sulfadiazine + pyrimethamine) in other to verify their viability by MTT or LDH assay, respectively. Next, BeWo cells were infected with T. gondii RH (2F1 clone) or ME49 strain, whereas villous were infected only with RH strain (2F1 clone), after, both cells and villous were treated or not with the same antibiotics and analyzed to T. gondii intracellular proliferation by beta-galactosidase assay (for RH strain) or blue toluidine staining (for ME49 strain). ELISA was performed in the supernatant to evaluate the cytokine profile. Enrofloxacin and toltrazuril did not change strongly the viability in cells and villous. Furthermore, the drugs decreased the parasite intracellular proliferation regardless T. gondii strain in BeWo cells and villous explants when compared to untreated and infected conditions. In BeWo cells infected by RH, enrofloxacin induced high levels of IL-6 low levels of MIF, while both cytokines were upregulated by enrofloxacin and toltrazuril in BeWo cells infected by ME49 strain. Additionally, in villous explantes, enrofloxacin induced high MIF production. Thus, enrofloxacin and toltrazuril were able to control the parasitism in BeWo cells and villous explants, and probably it occurs by modulation of immune response in these cells or tissues and direct action on parasite, but future experiments are necessary to verify this hypothesis.

Infección de transmisión vertical por Toxoplasma gondii: seguimiento de los hijos de mujeres con primoinfección en una institución de asistencia médica colectiva; 2010-2015

La toxoplasmosis congénita (TC) afecta 1 a 2 niños cada 1.000 nacimientos al año. La mayoría de los recién nacidos infectados son asintomáticos pero la ausencia de tratamiento puede determinar secuelas oftalmológicas y neurológicas. Objetivo: describir el seguimiento de los hijos de mujeres con primoinfección por Toxoplasma gondii durante el embarazo derivados a una Policlínica de Infectología de la Médica Uruguaya entre diciembre de 2010 y mayo de 2015. Material y método: se incluyeron los hijos de mujeres con primoinfección por T.gondii durante el embarazo entre el 1 de diciembre de 2010 y el 31 de mayo de 2015. Se confirmó primoinfección mediante determinación inmunoenzimática de IgG e IgM específicas, complementada por IgM por inmunofluorescencia indirecta o test de avidez de IgG según el caso. El diagnóstico de infección congénita se realizó por la presencia de IgM o títulos de IgG estables o en aumento en los primeros 9 meses de seguimiento del niño. Resultados: se diagnosticó primoinfección en 34 mujeres. La mayoría controló adecuadamente el embarazo y ninguna presentó infección por VIH, sífilis o Chagas. Se confirmó TC en 3 niños nacidos a término, con peso adecuado, hijos de mujeres con primoinfección adquirida en el tercer trimestre y tratadas con espiramicina. Uno presentó coriorretinitis, los otros fueron asintomáticos. En todos la IgM fue negativa, el diagnóstico se confirmó con curva de IgG. Todos recibieron piremetamina, sulfadiazina y ácido folínico sin efectos adversos. A la fecha continúan en tratamiento y seguimiento dos de los tres niños. Discusión y conclusión: la captación temprana de la mujer embarazada, la indicación oportuna de medidas de prevención constituyen pilares fundamentales para reducir la TC. El tratamiento oportuno y adecuado puede prevenir las secuelas.

Fever, Hemiparesis and Hyponatraemia in a 63-year-old. HIV, another Great Masquerader?

The authors describe the unusual case of a 63-year-old patient who was referred with fever and lethargy, and was found to be hyponatraemic. The patient subsequently developed hemiparesis, and neuroradiology showed several space-occupying brain lesions. The cause was later identified as cerebral toxoplasmosis in undiagnosed Acquired Immunodeficiency Syndrome (AIDS).

Cerebro-meningeal infections in HIV-infected patients: a study of 116 cases in Libreville, Gabon.

Background: Cerebro-meningeal pathology is common in human immunodeficiency virus (HIV) infection and the aetiology is often difficult to ascertain with certainty. Objective: To describe the major suspected and identified causes of meningeal or encephalitic syndromes in HIV infection in Libreville, Gabon. Methods: A descriptive study using clinical records of patients hospitalised in the Department of Medicine in the Military Hospital of Libreville (Gabon) between January 2006 and May 2010. Clinical features were evaluated using multivariable logistic regression to evaluate association with the outcome of a clinical improvement or death. Results: The most frequent neurological symptoms were reduced level of consciousness (54.3%), headache (55.2%), motor deficit (38.7%), and convulsions (36.2%). Cerebral toxoplasmosis represented 64.7% of diagnoses, followed by cryptococcal neuromeningitis in 12.9% of cases. Tuberculoma was diagnosed in 4 cases and lymphoma in 2 cases. In 9.5% of cases, no aetiology was determined. Toxoplasmosis treatment led to clinical improvement in 69.3% of cases with suspected cerebral toxoplasmosis. Overall mortality was 39.7%. Conclusion: The diagnosis of neurological conditions in HIV positive patients is difficult, particularly in a low-resource setting. A trial of treatment for toxoplasmosis should be initiated first line with all signs of neurological pathology in a patient infected with HIV.

Análisis de la capacidad diagnóstica de zoonosis neurológicas de tipo viral en équidos en Cundinamarca

Tesis (Médico Veterinario). -- Universidad de La Salle. Facultad de Ciencias Agropecuarias. Programa de Medicina Veterinaria, 2014

Determinación de la prevalencia serológica de brucella abortus, toxoplasma gondii, sarcocystis spp y los factores de riesgo asociados en estudiantes de medicina veterinaria de la Universidad de la Salle

Tesis (Médico Veterinario). -- Universidad de La Salle. Facultad de Ciencias Agropecuarias. Programa de Medicina Veterinaria, 2013

Plan estratégico para el control de roedores y palomas en la corporación de abastos de Bogotá S.A.

Tesis (Médico Veterinario). -- Universidad de La Salle. Facultad de Ciencias Agropecuarias. Programa de Medicina Veterinaria, 2014

Pathogenicity and phenotypic sulfadiazine resistance of Toxoplasma gondii isolates obtained from livestock in northeastern Brazil

Toxoplasma gondii is the causative protozoan agent of toxoplasmosis, which is a common infection that is widely distributed worldwide. Studies revealed stronger clonal strains in North America and Europe and genetic diversity in South American strains. Our study aimed to differentiate the pathogenicity and sulfadiazine resistance of three T. gondii isolates obtained from livestock intended for human consumption. The cytopathic effects of the T. gondii isolates were evaluated. The pathogenicity was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using a CS3 marker and in a rodent model in vivo. Phenotypic sulfadiazine resistance was measured using a kinetic curve of drug activity in Swiss mice. IgM and IgG were measured by ELISA, and the dihydropteroate synthase (DHPS) gene sequence was analysed. The cytopathic effects and the PCR-RFLP profiles from chickens indicated a different infection source. The Ck3 isolate displayed more cytopathic effects in vitro than the Ck2 and ME49 strains. Additionally, the Ck2 isolate induced a differential humoral immune response compared to ME49. The Ck3 and Pg1 isolates, but not the Ck2 isolate, showed sulfadiazine resistance in the sensitivity assay. We did not find any DHPS gene polymorphisms in the mouse samples. These atypical pathogenicity and sulfadiazine resistance profiles were not previously reported and served as a warning to local health authorities.

Ocorrência da infecção por Toxoplasma gondii e avaliação da imunização em caprinos do sertão do cabugi, Rio Grande do Norte

Toxoplasmosis is one zoonosis caused by Toxoplasma gondii protozoan. Goats, amongst the production animals, are one of the species most susceptible to this parasite, being one them main involved agents in ovine and goat abortions, determining great economic losses and implications for public health, since the presence it parasite in the products of goat origin, consist in one of the main sources of infection for the man. In this study 244 blood samples in 8 farms situated in 4 cities from the Sertão do Cabugi region, Rio Grande do Norte State, northeast of Brazil and, tested by ELISA assay. The results had shown a prevalence of 47.13% for anti- T. gondii antibodies and a significant association between positivity and variable evaluated as age, locality and property. The IgG avidity assay evaluated in 115 positive samples was carried to discriminate acute and chronic infection. Twelve samples (10.4%) had presented antibodies of low avidity while 103 (89.6%) presented high avidity antibodies; indicating that most of the animals was precocious exposure to the parasite. Significant difference was verified only for the variable sex. We also evaluate the capacity of recombinant adenoviruses codifying SAG1, SAG2, SAG3 and CMV in inducing activation of specific immune response in goat. These 109 animals received 109 pfu of the AdSAG1, AdSAG2, AdSAG3, AdCMV or PBS in vaccine protocol with 3 immunizations. Serum samples of the each animal, before and after mmunization, had been submitted to the ELISA. The results demonstrate that the immunizations had induced the production of IgG antibodies specific against T. gondii proteins

Oral administration of encapsulated bovine lactoferrin protein nanocapsules against intracellular parasite Toxoplasma gondii

Toxoplasma gondii is a deadly intracellular parasite known to reside in every nucleated cell and known to cause severe complications in immunocompromised host. Standard drugs are cost effective and cause side effects, therefore, there is a necessity for a new drug molecule with immunomodulatory potential. Lactoferrin (Lf) is a natural milk protein, which has shown antimicrobial properties in its nanoformulation using alginate chitosan calcium phosphate bovine lactoferrin nanocapsules (AEC-CCo-CP-bLf-NCs). The present study was aimed to analyze and compare the effect of bovine Lf (bLf) in its native as well as nanoformulation (AEC-CCo-CP-bLf-NC) against coccidian parasite T. gondii. In vitro analysis has shown a significant increase in nitric oxide production and low parasitemia in in vitro cell culture model. In vivo BALB/c mice model have been used to develop human toxoplasmosis model. After treatment with NCs it has substantially increased the bioavailability of the protein and showed comparatively increased levels of reactive oxygen species, nitric oxide production, and Th1 cytokine which helped in parasite clearance. The mechanism of action of NCs has been clarified by immunoreactivity analysis, which showed accumulation of Lf in macrophages of various visceral organs, which is the site of parasite multiplication. Effect of NCs has significantly decreased (P<0.05) the parasite load in various organs and helped survival of mice till day 25 postinfection. Fe metabolism inside the mice has been found to be maintained even after administration of mono form of Lf, this indicates novelty of Lf protein. From the present study we concluded that nanoformulation did not reduce the therapeutic potential of Lf protein; however, nanoformulation has enhanced the stability of the protein and shown anti-toxoplasmal activity. Our study presents for the first time nanoformulation of Lf protein against Toxoplasma, which has advantages over the standard drug therapy without any side effects.