First molecular-based detection study of Leishmania infantum in the Tapirapé indigenous population in the Brazilian Amazon.

Species of the genus Leishmania parasitize mammals and have life cycles that alternate between vertebrate and invertebrate hosts. Most species develop in a hematophagous arthropod and infect a specific vertebrate host that may belong to diverse orders and families. Visceral leishmaniasis is a chronic zoonosis with a wide geographic distribution, affecting 350 million people globally, mostly in areas with a high risk of infection. In Brazil, this disease not only has a high incidence but is also expanding to new areas, both in urban centers and rural areas, including territories with tribal communities, due to increasing human intervention. The objective of this study was to perform cathepsin L-like gene-based molecular diagnosis of Leishmania infantum in the indigenous Tapirapé ethnic group in the state of Mato Grosso. From the 372 individuals assessed, only 0.8% (3/372) tested positive for L. infantum, all from the same village (Urubu Branco). Despite the small number of infected individuals, this study demonstrates the first human cases of Leishmania infantum infection in this population, suggesting the need for regular monitoring of visceral leishmaniasis in the area and leading to a broad discussion on the planning and implementation of public health measures for the indigenous population, while respecting their distinctive territories and culture.

First molecular-based detection study of Leishmania infantum in the Tapirapé indigenous population in the Brazilian Amazon

Species of the genus Leishmania parasitize mammals and have life cycles that alternate between vertebrate and invertebrate hosts. Most species develop in a hematophagous arthropod and infect a specific vertebrate host that may belong to diverse orders and families. Visceral leishmaniasis is a chronic zoonosis with a wide geographic distribution, affecting 350 million people globally, mostly in areas with a high risk of infection. In Brazil, this disease not only has a high incidence but is also expanding to new areas, both in urban centers and rural areas, including territories with tribal communities, due to increasing human intervention. The objective of this study was to perform cathepsin L-like gene-based molecular diagnosis of Leishmania infantum in the indigenous Tapirapé ethnic group in the state of Mato Grosso. From the 372 individuals assessed, only 0.8% (3/372) tested positive for L. infantum, all from the same village (Urubu Branco). Despite the small number of infected individuals, this study demonstrates the first human cases of Leishmania infantum infection in this population, suggesting the need for regular monitoring of visceral leishmaniasis in the area and leading to a broad discussion on the planning and implementation of public health measures for the indigenous population, while respecting their distinctive territories and culture.

Allometric coefficients of major chemical components of meat quail raised in different thermal environments.

The objective of the present study was to estimate and compare allometric coefficients of the major chemical components of meat quail raised in different thermal environments, based on protein weight of feather free body (FFB) and feathers. In total, 300 meat quail, males and females, were distributed in a completely randomized design with 2 treatments (climatized environment, 26°C, and non-climatized environment, 29°C) and 6 replicates of 25 birds each. On the first day, 36 birds were selected to form the reference group and from this day on, 2 quail were weekly sampled from each cage. All selected birds were fasted for 24 h, weighed, slaughtered, plucked, and reweighed. The FFB and feathers were ground separately to obtain homogeneous samples, which were freeze-dried to determine the water content, and thereafter, ground again in a micromill before analyzing for protein, lipid, and ash using AOAC procedures. The adjustment of the allometric equations was made using crude protein (CP) weight as the independent variable and water, lipid, and ash weight as the dependent variables. The data of each dependent variable were transformed into natural logarithm (ln), regressed according to lnCP, and subjected to a parallelism test. In the FFB, water showed early development and lipid and ash showed late development in relation to the CP weight. In feathers, water and ash weight showed early development in relation to the feathers protein weight, whereas lipids showed late development. The environments of 26°C and 29°C did not affect the allometric coefficients that described the growth between the chemical components in the body and in the feathers, except for lipids in male FFB, that showed higher allometric coefficient at 29°C than 26°C. Describing the allometric relationships between the major chemical components of meat quail body is an important step in supporting future research comprised modeling of body growth and nutrition for meat quail.

Prevalência de anticorpos anti-Rickettsia spp. em cães da cidade de Belo Horizonte, MG / Prevalence of antibodies anti-Rickettsia in dogs from Belo Horizonte, MG, Brazil

A cross-sectional study was conducted to determine the seroprevalence of anti-Rickettsia spp. in dogs from Belo Horizonte, MG. For this purpose, serum samples from 453 dogs were collected during the rabies vaccination campaign and tested by the indirect immunofluorescence assay (IFA) using five antigens: Rickettsia bellii, Rickettsia amblyommii, Rickettsia rickettsii, Rickettsia parkeri, and Rickettsia felis. The endpoint titer reacting with each antigen was determined and serum was considered positive if it reacted at the 1:64 dilution. Serum showing titer for a Rickettsia species at least four-fold higher than that observed for any other Rickettsia species was considered homologous to the first Rickettsia species. Only three (0.66%) dogs reacted positively to at least one Rickettsia species and one serum showed to be homologous to R. rickettsii. These results showed a low prevalence of antibodies anti-Rickettsia spp. in dogs from Belo Horizonte city. However, other serosurvey needs to be performed for surveillance of the endemic status of the disease in the municipal district.

Purification and biochemical characterization of Eumiliin from Euphorbia milii var. hislopii latex.

A protease, which we designate Eumiliin, was isolated from the latex of Euphorbia milii var. hislopii by a combination of ion-exchange chromatographic steps using DEAE-Sephacel and gel-filtration with Sephadex G-75. Eumiliin is a monomeric protein with an apparent molecular mass of 30 kDa by SDS-PAGE under reducing conditions and gave one main peak at 29,814 KDa in MALDI-TOF/TOF mass spectrometry. Eumiliin has caseinolytic and fibrinogenolytic activities, but no hemorrhagic or defibrinating activities. The enzyme readily hydrolyzes the Aalpha-chain of fibrinogen and, more slowly, the Bbeta-chain. Its fibrinogenolytic activity is inhibited by beta-mercaptoethanol and leupeptin. In contrast, EDTA and benzamidine did not affect the activity of Eumiliin. The caseinolytic activity of Eumiliin had a pH optimum of 8.0 and was stable in solution at up to 40 degrees C; activity was completely lost at >or=80 degrees C. Intraplantar injection of Eumiliin (1-25 microg/paw) caused a dose- and time-dependent hyperalgesia, which peaked 1-5h after enzyme injection. Intraplantar injection of Eumiliin (1-25 microg/paw) also caused an oedematogenic response that was maximal after 1h. Morphological analyses indicated that Eumiliin induced an intense myonecrosis, with visible leukocyte infiltrate and damaged muscle cells 24h after injection.

An alternative method for DNA extraction and PCR identification of Entamoeba histolytica and E. dispar in fecal samples.

Since it is known that Entamoeba dispar is non-pathogenic and morphologically similar to E. histolytica, there are many targets used in PCR for differentiating these species. However, obtaining high quality DNA from fecal samples is fundamental for PCR. Most methods are laborious or use kits that make diagnosis expensive. In the present work, a new simple, fast and cheap technique of DNA extraction from fecal samples was combined with a PCR for an episomal target in order to identify E. histolytica and E. dispar in feces.

Resuscitative procedures at birth in late preterm infants.

OBJECTIVE: Evaluate the need for resuscitative procedures at birth, in late prematures. STUDY DESIGN: This prospective cohort study enrolled all liveborn infants from 1 to 30 September 2003, with 34 to 41 weeks of gestation without congenital anomalies, born in 35 public hospitals of 20 Brazilian state capitals. Logistic regression analyzed variables associated with the need for bag and mask ventilation. RESULT: Of the 10 774 infants studied, 1054 were late preterms and 485 required resuscitative measures. Of the 1054, 338 (32%) received only free-flow oxygen, 143 (14%) were bag and mask ventilated, 27 (3%) were intubated and 10/27 received chest compressions and/or medications. Bag and mask ventilation in late preterms was associated with twin gestation, maternal hypertension, nonvertex presentation, cesarean delivery and lower gestational age. CONCLUSION: Improving control of maternal hypertension, prolonging gestation for 1 to 2 weeks and restricting operative deliveries could decrease the need of resuscitation of late preterms at birth.

Effects of glycerol on the thermal dependence of the stability of human erythrocytes.

Incubation of human blood in saline solution of 0-36% (v/v) ethanol for 30 min produces lysis or stabilization of erythrocytes depending on the ethanol concentration. Under less elevated concentrations of ethanol, erythrocytes are present in expanded shapes (R state) that present lower stability and suffer lysis with increase in the ethanol concentration. Under more elevated concentrations of ethanol, erythrocytes are present in contracted shapes (T state) that have higher stability and suffer lysis at even more elevated ethanol concentrations. This work evaluated the effects of glycerol (0 to 2.0 M) and temperature (7 to 47 degrees C) on the stability of the R erythrocytes, characterized by the ethanol concentration at the mid-transition point (D (50R )) of the hemolysis curve (D (50R )). D (50R ) declined sigmoidally with increase in the glycerol concentration or temperature, due to transition of the R to the T state erythrocytes. In 1.5 M glycerol, the erythrocytes stability decreased below 32 but increased above 37 degrees C. The combination of temperature, glycerol and ethanol actions generates a critical value of osmotic pressure below which the R state predominates and above which the T state predominates. At 7 degrees C 1.5 M glycerol decreased the erythrocytes stability against ethanol but increased the erythrocytes stability against hypotonic shock. Those conditions favor the R state, which has a lower stability against ethanol; however, in the absence of ethanol, glycerol determines less water entrance in the erythrocytes, making more difficult its lysis by hypotonicity.

Purification and functional characterization of two fibrinogenolytic enzymes from Bothrops alternatus venom

Two fibrinogenolytic enzymes, Bothrops alternatus metalloprotease isoform (BaltMP)-I and II, were purified from Bothrops alternatus venom using Diethylaminoethyl (DEAE) Sephacel, Sephadex G-75 and Heparin-Agarose column chromatography. Purified BaltMP-I and II ran as single protein bands on analytical polyacrylamide gel electrophoresis and showed molecular weights of 29000 and 36000, respectively, under reducing conditions in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). BaltMP-II, but not BaltMP-I, displayed blood-clotting activity in bovine plasma, which was about 10-fold higher than that of the crude venom. Both enzymes were proteolytically active against bovine fibrinogen as substrate. When fibrinogen and each enzyme were incubated at 37°C, at a ratio of 1:100 (w/w), BaltMP-II cleaved preferentially the Aalpha -chain and more slowly the Bbeta -chain. The action of BaltMP-I was similar, but lower. None of the proteases degraded the gamma-chain of fibrinogen. The fibrinogenolytic activity of the enzymes was inhibited by 1,10-phenanthroline, suggesting they are metalloproteases. Since both enzymes were found to cause defibrinogenation when intraperitoneally (i.p.) administered to mice, they can be of medical interest as a therapeutic agent in the treatment and prevention of arterial thrombosis.(AU)

Atividade antimicrobiana de bactérias lácticas de embutidos curados frente a Listeria monocytogenes / Antimicrobial activity of lactic acid bacteria isolated from Brazilian dry fermented sausages against Listeria monocytogenes

Cepas de bactérias lácticas recuperadas de 336 colônias isoladas e selecionadas foram submetidas ao teste de atividade antimicrobiana direta, que identificou as produtoras de substâncias antimicrobianas capazes de inibir in vitro o desenvolvimento de duas cepas indicadoras de Listeria monocytogenes. As 108 cepas que inibiram diretamente pelo menos uma das cepas indicadores receberam a denominaçäo DTEI e foram selecionadas para o teste de atividade antimicrobiana indireta contra as mesmas cepas de L. monocytogenes, assim como frente a outras cepas de bactérias lácticas de origens diversas. Essa atividade inibidora indireta foi avaliada por meio de sobrenadantes isentos de células, esterilizados por meio de microfiltraçäo, eliminando-se os principais compostos responsáveis por ela, como por exemplo os ácidos orgânicos e o peróxido de hidrogênio, mediante o ajuste do pH e a liofilizaçäo dos sobrenadantes. Oito cepas de bactérias lácticas apresentaram atividade antimicrobiana indireta frente a pelo menos um dos microrganismos indicadores utilizados, sugerindo terem produzido substâncias semelhantes a bacteriocinas. Três destas cepas foram caracterizadas e identificadas como pertencentes ao gênero Lactobacillus sp

Epidemiological aspects of Babesia equi in horses in Minas Gerais, Brazil.

The prevalence of Babesia equi in two climatic regions of Minas Gerais state was determined using the indirect fluorescent antibody test (IFAT) with blood samples obtained from horses in two slaughterhouses. Of 399 samples, 241 (60.4%) showed a positive reaction. Anti-B. equi antibody was detected in every county studied, the prevalence being 59.7% for horses in the area where the temperature rises above 18 degrees C in winter and 61.4% in the area where it remains below 18 degrees C, indicating that climatic variation has no substantial effect on the prevalence of the infection in Brazil. Blood samples collected from all 95 horses on a ranch in the state of Minas Gerais, Brazil, on which clinical babesiosis had never been reported, were subjected to the IFAT Anti-B. equi antibodies were detected in horses of all ages, but with a significantly lower prevalence in animals less than 6 months old.

Neutralizing potency of horse antibothropic antivenom. Correlation between in vivo and in vitro methods.

The correlation coefficients between in vivo neutralization of lethal toxicity (ED50), neutralization of the hemolytic activity (PLA2) and levels of antibodies measured by ELISA, was investigated to test the potency of horse anti-bothropic antivenom. Twenty six horses were hyperimmunized with Bothrops venoms (B. alternatus, B. jararaca, B. jararacussu, B. neuwiedii and B. moojeni). To set up an indirect ELISA, for neutralization of PLA2 activity and for determination of ED50 in Swiss mice, the whole Bothrops jararaca venom (reference venom for assessing the bothropic antivenom potency in Brazil) was used. The toxic fraction (purified from B. jararaca venom by Sephadex G-100 chromatography) was also used as antigen for ELISA. All antivenoms analyzed effectively neutralized the lethal activity in the range of 1.6 to 9.6 mg/ml of antivenom. The correlation coefficient between ED50 and ELISA antibody titers against the crude venom and toxic fraction was r = 0.65 (P < 0.001) and r = 0.85 (P < 0.0001), respectively. Correlation between ED50 and neutralization of PLA2 activity was r = 0.52 (P < 0.01), and the correlation between ELISA antibody titers and neutralization of PLA2 activity was r = 0.58 (P < 0.002). Thus, the ELISA which measures only the antibody against the major toxic fraction of the B. jararaca venom should be most suitable for use as an in vitro assay of bothropic antivenom potency.

Estudo anátomo-patológico de cäes infectados experimentalmente pelo Angiostrongylus vasorum (Baillet, 1866) Kamenski 1905 / Angiostrongylus vasorum (Baillet, 1886) Kamenski 1905: pathological manifestations in experimentally infected dogs

Dez cäes mestiços, dois mantidos como controle e oito infectados experimentalmente com uma amostra de Angiostrongylus vasorum, foram usados para estudar a infecçäo experimental pelo parasito. Caramujos da espécie Biomphalaria glabrata, criados em laboratório, serviram como hospedeiros intermediários para o parasito. Aos seis meses de idade os animais foram inoculados com diferentes doses de larvas infectantes de A. vasorum, e a cada 30 dias foram sacrificados dois cäes (do 86§ ao 210§ dia após a inoculaçäo). Como o hábitat do A. vasorum é a artéria pulmonar e o átrio direito, as lesöes mais freqüentes e severas foram observadas nos pulmöes dos animais inoculados. Sufusöes hemorrágicas e hematomas foram encontrados no cérebro, no coraçäo, nos pulmöes, no estômago e no rins. Parasitos adultos, além de numerosos ovos e larvas de A. vasorum, foram também encontrados em vasos sangüíneos do pâncreas, do coraçäo e dos linfonodos, formando verdadeiros "ninhos" no coraçäo, rins, pâncreas e linfonodos. Foi observado pela primeira vez que o A. vasorum, independente de sua presença no tecido pancreático, pode causar pancreatite crônica difusa inespecífica, pancreatite crônica produtiva granulomatosa difusa e arterite crônica com trombose ocludente. No coraçäo os adultos causam endarterite obliterante, enquanto os "ninhos" de ovos e larvas provocam o desenvolvimento de miocardite crônica difusa granulomatosa, cuja confluência tem a aparência macroscópica de enfarte do miocárdio. Fibrilopoiese difusa, näo relacionada diretamente com os focos inflamatórios, nos diferentes órgäos foi observada em dois casos. Nos rins, a presença de parasitos estava associada com glomerulonefrite mesângio-proliferativa difusa, com granulomas confluentes e tromboses arteriais múltiplas

Control of schistosomiasis mansoni in Ravena (Sabará, state of Minas Gerais, Brazil) through water supply and quadrennial treatments.

In this study, the results obtained in a control programme of schistosomiasis in Ravena (Sabará, Minas Gerais) between 1980 and 1992 are evaluated. Control measures used in this programme were: specific treatment of the people infected with Schistosoma mansoni at four year-intervals (1980/84/88) and the supply of tap water to 90% of the residences in 1980. A significant reduction of the prevalence (36.7% to 11.5%, p < or = 0.05) and of the intensity of the infection (228.9 eggs per gram of feces (epg), s = 3.7 to 60.3 epg, s = 3.5, p < or = 0.05) was observed. No cases of the severe form of the disease were diagnosed in the area. Factors independently associated with the infection were in 1980 daily sand extraction and the lack of tap water in residences and in 1992 daily sand extraction and fishing and weekly swimming. Concluding, the supply of tap water together with quadrennial treatments significantly diminished both the prevalence and intensity of the S. mansoni infection, with the additional gain of persistent low indices even after four-year intervals between the treatments.

Activity of 9-acridanone-hydrazone drugs detected at the pre-postural phase, in the experimental schistosomiasis mansoni.

The compound Ro-15.5458/000, derivative in the class of 9-acridanone-hydrazones, was found to be effective against Schistosoma mansoni in mice, killing almost all the skin schistosomules (24 hr after infection), when administered at the dose of 100 mg/kg. In experiments carried out with Cebus monkeys, the drug was shown to be fully effective at 25 mg/kg, 7 days after infection. These data, associated with the good results obtained earlier at the post-postural phase of schistosomiasis, allow the inference that this promising compound may be important in the set of antischistosomal drugs, depending on further toxicological and clinical tests.

Effective immune protection of pigs against cysticercosis.

A scolex protein antigen (SPA) was prepared from cysticerci of Taenia solium obtained from naturally infected pigs. Yorkshire pigs were vaccinated with SPA plus incomplete Freund's adjuvant (IFA) or with SPA plus Corynebacterium parvum (CP). Controls were given IFA plus phosphate-buffered saline (PBS) or CP plus PBS. All animals were given three subcutaneous injections at 20-day intervals. Ten days after the third injection, the pigs were fed with 10(4) viable eggs of T. solium. All pigs developed a delayed type hypersensitivity, and a transient eosinophilia after the first dose of vaccine. High titers of specific antibodies were detected in the sera of vaccinated animals and in infected controls. A protection level of 71.43% was recorded in animals vaccinated with SPA plus IFA and of 75.00% in those vaccinated with SPA plus CP.

Efeito de infecçöes experimentais por Ascaris suum e Isospora suis em suínos da raça Piau / Effect of experimental infection with Ascaris suum and Isospora suis in swine \"Piau\" breed

Vinte e sete leitöes da raça Piau foram divididos em seis grupos e inoculados, por via oral, com oocistos de Isospora suis e ovos de Ascaris suum. Os leitöes dos Grupos I e II, cada um com seis animais, receberam, aos 10 dias de idade, respectivamente, 100.000 oocistos de Isospora (Grupo I) e 50.000 oocistos + 500 ovos de Ascaris (Grupo II). Os leitöes dos demais grupos, aos 45 dias de idade, foram submetidos aos seguintes tratamentos: Grupo III - Quatro animais inoculados com 100.000 oocistos de Isospora; Grupo IV - Quatro animais inoculados com 50.000 oocistos + 500 ovos de Ascaris; Grupo V - Quatro animais inoculados com 500 ovos de Ascaris; Grupo VI - Três animais mantidos como testemunhas. A infecçäo por Isospora suis somente tornou-se patente em leitöes inoculados aos 10 dias de idade. Os Grupos I e II apresentaram 85 por cento de mortalidade, provavelmente provocada por um surto de salmonelose agravado pela inoculaçäo de Ascaris e Isospora. Os leitöes inoculados aos 45 dias de idade pelos mesmos parasitos infectaram-se somente com Ascaris suum. A diferença de ganho de peso final e inicial, respectivamente aos 126§ e 42§ dias de idade, foi menor nos animais inoculados com Ascaris suum

Transmissäo congênita de Babesia bovis: relato de um caso autóctone em Minas Gerais - Brasil / Prenatal infection of calf with Babesia bovis: first autoctonous case in Brazil

It is discussed first autoctonous prenatal case of Babesia bovis in Brazil. After a normal birth a male Brown Swiss calf, had not sucked colostrum and died at three days after birth. Clinical and laboratory examinations revealed petechial hemorrhages in oral and conjuntival mucosas, red urine and parasitemia due to B. bovis as shown in blood smears Giemsa stained. Post-mortem examination revealed hepato-splenomegaly, congestion and edema in lungs, brain and kidneys. Optical and eletronic microscopies showed severe congestion, edema, red cells aglutination, and most of the erythrocites in the brain capillaries contained Babesia organisms, which were also detected outside the cells. In the kidneys interlobular interstitial nephritis, glomerulonephritis mesangioproliferative diffuse and rare parasites in the capillaries were observed. 1.8 per cent of the erythrocites in peripheral circulation were parasitized in the brain capillaries more than 90 per cent of those cells had Babesia organisms