Comparação de Meios de Enriquecimento e de Plaqueamento Utilizados na Pesquisa de Salmonella em Carcaças de Frango e Fezes de Aves

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Recent Studies on Neoechinorhynchus curemai Noronha, 1973 (Acanthocephala: Neoechinorhynchidae), in Prochilodus lineatus Valenciennes, 1836, from Volta Grande Reservoir, MG, Brazil

O presente trabalho estudou a helmintofauna do curimbatá, Prochilodus lineatus Valenciennes, 1836, do reservatório de Volta Grande, MG, Brasil. Foram analisados 18 peixes com comprimento médio de 46,7 ± 1,1 cm e peso médio de 1.674,8 ± 75,6 g, sendo que 15 apresentaram acantocéfalos no intestino com prevalência de 83,3%. O helminto foi identificado como Neoechinorhynchus curemai Noronha, 1973 (Acanthocephala: Neoechinorhynchidae), que diferiu das outras espécies descritas pelas dimensões dos caracteres e pela morfologia. da descrição original de N. curemai difere pelas maiores dimensões dos testículos, pela glândula de cimento alongada, pela presença de núcleos nos lemniscos, pelas dimensões dos ovos e pelos maiores ganchos da probóscide presentes na segunda e na terceira fileiras nos machos e na primeira fileira nas fêmeas. Foi observada menor porcentagem ocupada pelo sistema reprodutivo em relação ao tronco da fêmea. A observação dos parátipos de N. curemai de Noronha (1973) mostrou grande semelhança com os do presente trabalho. Este fato complementa a descrição do helminto em outra localidade.

Toxoplasma gondii: cloning, sequencing, expression, and antigenic characterization of ROP2, GRA5 and GRA7

Toxoplasma gondii is an intracellular obligate protozoan, which infects humans and warm-blooded animals. The aim of the present study was to clone the rop2, gra5 and gra7 genes from T. gondii RH strain and to produce recombinant proteins. The rop2, gra5 and gra7 gene fragments produced by polymerase chain reaction were cloned into the pET102/D-TOPO(R) vector which contains thioredoxin and polyhistidine tags at the C-and N-ends, respectively, and is expressed in Escherichia coli BL21(DE-3). The expression fusion proteins were found almost entirely in the insoluble form in the cell lysate. These recombinant proteins were purified with an Ni-NTA column. Concentrations of the recombinant antigens produced in the E. coli BL21-star ranged from 300 to 500 mu g/mL growth media, which was used to immunize rabbits. We observed an identity ranging from 96 to 97% when nucleotide sequences were compared to GenBank database sequences. Immunocharacterization of proteins was made by indirect immunofluorescence assay. These proteins will be used for serodiagnosis and vaccination.

Transgenic lettuce seedlings carrying hepatitis B virus antigen HBsAg

The obtainment of transgenic edible plants carrying recombinant antigens is a desired issue in search for economic alternatives viewing vaccine production. Here we report a strategy for genetic transformation of lettuce plants (Lactuca sativa L.) using the surface antigen HBsAg of hepatitis B virus. Transgenic lettuce seedlings were obtained through the application of a regulated balance of plant growth regulators. Genetic transformation process was acquired by cocultivation of cotyledons with Agrobacterium tumefaciens harboring the recombinant plasmid. It is the first description of a lettuce Brazilian variety Vitória de Verão genetically modified.

Eficácia anti-helmíntica comparativa da associação albendazole, levamisole e ivermectina à moxidectina em ovinos

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Avaliação de dois clones de Escherichia coli recombinante quanto ao crescimento e expressão de antígenos de Leishmania chagasi (kmp11 e P36)

Visceral leishmaniosis caused by Leishmania chagasi, also known as calazar, presented, in the period from 1990 to 2005, tax of incidence in Brazil varying between 1 and 3 cases for 100 000 inhabitants. The Northeast region that up to the year of 2000 contributed with almost 90% of the registered cases is reducing his participation in the current decade, reaching 56% in 2005. Conventional leishmaniasis treatment is costly and it shows high toxicity, demanding more research for alternative treatments, with special interest in development of vaccines and diagnosis kits which include production of recombinant antigens by host cells. Escherichia coli has been the microorganism most studied and used as a host for recombinant protein production. Therefore, the aim of this work was to study the influence of induction on cellular growth and to verify the type of Leishmania chagasi antigens expression (intra or extracellular) during two recombinant E. coli clones (kmp11 and P36) cultivation in rotary incubator (shaker) using three different media (2xTY, TB, FASS+EL). For that, tests were carried out using conditions established in the literature for E. coli (37°C and 200 rpm) and media supplemented with antibiotics to guarantee that only competent cells grows. First, tests were carried out without induction in order to verify the two microorganisms kinetic behavior (growth and substrate consumption) in different media. Next, the induction was carried out through the addition of IPTG (1mM as final concentration), at the first hour of cultivation. It was observed that protein expression were intracellular for all clones and media tested, however the highest level of expression was clearly observed by the electrophoresis band density (intensity) for 2xTY medium and kmp11 protein. Although it contains the lowest substrate concentration, consequently, a reduced cellular concentration when compared to other media, it appeared that this medium and clone combination is the most indicated for recombinant protein production. Therefore, the objective of this work was achieved, since the interested proteins were produced. Consequently, this result motivates new studies for production optimization using different cultivation strategies

Influência das condições de cultivo na produção de antígenos recombinantes de Leishmania i. chagasi utilizando Escherichia coli M15 cultivada em incubador rotativo e biorreator

Escherichia coli has been one of the most widely used hosts in recombinant protein production, in both laboratory and industrial scale since the advent of recombinant DNA technology. Despite the substantial progress of studies on the molecular biology and immunology of infections, there is currently no medication-based prophylaxis capable of preventing leishmaniasis. As such, there is a great need to identify specific antigens for the development of vaccines and diagnostic kits against visceral leishmaniasis. Thus, the primary goal of the present study is to assess the influence of cultivation conditions on the production of Leishmania chagasi antigens, carried out in a rotating incubator and bioreactor. To that end, several assays were conducted to evaluate the kinetic behavior of antigens (648, 503) of Leishmania. i. chagasi in two different compositions of media (2xTY, TB), with and without an inducer. In order to improve expression, assays were performed in a benchtop bioreactor using the best conditions obtained in a rotating incubator, in addition to assessing the influence of stirring speed. Results show that high complexity of the cultivation medium favored kinetic growth of clones (648, 503). However, in assays submitted to induction by IPTG, this elevated complexity did not promote the expression of recombinant proteins. Expression of antigens 648 and 503 exhibited behavior associated with growth and, in terms of location, proteins 648 and 503 are intracellularly stored. Lactose may be the most adequate inducer in protein expression, when considering factors, cost, toxicity and stability. Elevated stirring may increase cell growth in clone 53, although it may not result in high concentrations for the protein of interest. On the other hand, positive results were obtained for all recombinant clones (648, 503) tested, confirmed by the electrophoretic profile

Avaliação da eficiência de filtros anaeróbios na remoção de coliformes fecais e ovos de helmintos

The technology of anaerobic reactors for sanitary wastewater treatment has been extensively developed in Brazil, and today it is practically consolidated. They present several advantages, such as low construction and operating costs, and low sludge production, the anaerobic reactors are an attractive alternative to minimize problematic lack of basic sanitation in urban areas, and also of the rural areas. The anaerobic filters have been widely used in Brazil. It produces an effluent with low concentration of organic matter and solids suspended, besides conserving the nutrients, therefore, it is good for use in irrigation, but the practice must be associated with knowledge of the pathogens presence. The main objective of this study was to evaluate the efficiency of anaerobic filters in removal faecal coliforms and helminth eggs, and to verify if the effluent can be used for agricultural purposes, according to the World Organization of Health (WHO, 1989). The protocol used to enumerate helminths eggs was the modified Bailenger method, (Ayres and Mara, 1996) recommended by WHO for evaluation of raw effluent and treated effluent. The membrane filtration method was utilized to determine the concentrations of faecal coliforms. Three different systems of sewer treatment composed by anaerobic filters were analyzed. The results, in a general analysis, showed that all the researched systems reached a larger removal than 93% to helminth eggs, resulting in an effluent with smaller average than 1 egg/L. One of these systems, Sistema RN, reached a larger removal than 99%, confirming the good performance of the anaerobic filters in removal helminths eggs. Even with low concentrations of eggs in the influent, the filters were capable to remove this parameter efficiently. About faecal coliforms, it was observed for all the researched systems an effluent with 106 CFU/100mL. The high concentrations to faecal coliforms in the effluent just allow reuse for restricted irrigation, in agreement with the guidelines of WHO. Although the researched systems have not removed faecal coliforms efficiently, the results indicated a good efficiency of the anaerobic filters in removal helminth eggs

Caracterização dos resíduos esgotados de sistemas de tratamento individual de esgotos de Natal

In Natal still dominates the use of individual disposal systems for domestic sewage, once only 29% of the city has a sewarage system. Wastes that are accumulated in these individual treatment systems should be exhausted periodically, service provided by collector entrepreneurs. Some of these companies causing major damage to the environment. In Natal, only two companies have their own septage (RESTI) treatment system, which were designed with parameters from domestic sewage generating strain and inefficient systems. Therefore, the characterization becomes essential as a source of parameters for their design. Thus, this work presents the physical-chemical and microbiological characterization of waste pumped from individual sewage treatment systems. Samples collections were made weekly from 5 different trucks at the reception point on the treatment plant on the point of the preliminary treatment. From each truck it was taken 5 samples during the discharge in order to make a composite sample. Afterwards, samples were carried out to laboratory and analyses for determination of temperature, pH, conductivity, BOD, COD, nitrogen (ammonia e organic), alkalinity, oils, phosphorus, solids, faecal coliforms and helminth egg. The results were treated as a single database, and ranked according to its generating source (multi and single house, lodging, health, services and / or food), area of origin (metropolitan, south and north) and type of system (cesspits, septic tank and / or sink). Through these data it was possible to verify that the type of system adopted by most in Natal and the metropolitan region is cesspit, besides to confirm the difference between the septage of areas with a population have different social and economical characteristics. It was found that the septage have higher concentrations than domestic sewage, except for thermotolerant coliforms that showed concentrations of 1,38E+07. Among the parameters studied, is the median values identified for COD (3,549 mg / L), BOD (973mg / L) and total solids (3.557mg / L). The volatile fraction constitutes about 70% of the total solids of the septage. For helminths has been a median of 7 eggs/L. In general, the characteristics of the waste followed the variability found in the literature reviewed for all variables, showing high amplitudes

Protection induced in BALB/c mice by the high-molecular-mass (hMM) fraction of Paracoccidioides brasiliensis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Caracterização do conteúdo de fossas e tanques sépticos na cidade de Natal

Only 32% of the population of Natal is attended by sewage, while the remaining population use pits and septic tanks. The characterization of the contents of septic tanks and pits contributes to the performance of such system and may guide the decision on treatment of these contents. The main of this research is to characterize the contents of interior residential pits and septic tanks in the greater Natal, with the following specific goals: to develop and manufacture a sampler capable of collecting a representative sample of the entire column (the surface scum, the clarified liquid and sludge bottom); to compare the contents of the tanks with the pits; to compare the contents of the septage from vacuum trucks; to relate the composition content with socioeconomic characteristics of households; to compare the content in both chambers of the septic tanks in series; to assess the situation of the content before and six months after the cleanness; and ultimately propose a pilot scale plant for treatment of septage. Once the sampler was developed, samples were collected within 14 septic tanks and 10 pits in many districts of Natal. Medians of the 24 systems were obtained: temperature, pH, conductivity, oil and grease, total solids, total suspended solids and sediments of 28.0 °C, 6.95; 882 mS/cm, 75.2 mg/L; 10,169 mg/L, 6,509 mg/L and 175 mL/L respectively; 111.0 mgN/L for ammonia, 130.5 mgN/ L for organic nitrogen, 0.2 mgN/L for nitrite, 0.4 mg/L for nitrate; 8935 mgO2/L for COD, 29.2 mgP/L for total phosphorus, thermotolerant coliforms from 9.95 E +06 CFU/100mL helminth eggs and 9.2 eggs/L with a maximum concentration of 688 eggs/L and minimum of 0 eggs/L. Medians of organic nitrogen and TKN were significantly different between groups of tanks and pits. The systems with cleanness gap from 11 and 20 years presented the higher concentrations for most variables. The effluent from the toilets and bathrooms participate more effectively in contributing fractions of solids, alkalinity, nitrogen, COD, total phosphorus, thermotolerant coliforms and helminth eggs. The systems used by socioeconomics class with income from R$ 3,700.00 to R$ 7,600.00, presented higher concentrations for COD, nitrogen, solids and helminth eggs. The first of the two chambers had always presented higher concentrations over the second compartment. The analysis of variance for most variables, showed that the values of septic tanks, pits and septage from vacuum trucks belong to the same group. In the samples taken after cleanness, the median of pH and temperature increased, while alkalinity, COD, organic nitrogen, total phosphorus, ammonia and helminth eggs decreased. The oils and greases and thermotolerant coliforms had slightly varied due to the continuous release of sewage into the systems that maintained their steady state concentrations.

Uso urbano não potável de água de lagoas do sistema de drenagem de Natal

The detention and infiltration ponds of urban drainage system has function to protect the population from undesirable effects of floods. In general these ponds are not completly used and it potential is wasted. As it are disseminated at different places at cities it can reduce costs with water transport and permit water preservation of best quality. Some it utilities includes use in green areas irrigation, industrial among than cooling towers and boiler, soil compaction, cleaning urban road, pisciculture and fire fighting system. The quality these water is influenced by anauthorized sewage in the drainage system. This study was performed in six detention and infiltration ponds at drainage system of Natal, which aim was to verify the non-portable use these ponds on urban environment. As indirect aim would to incentive the water utilization these ponds as a water source to another uses in urban environment. These ponds represents the characteristics of detention and infiltration of Natal city and consequently of Brazil. As the water quality, the ponds presents following characteristics: three has apparently good quality, other showed intermediate condition and two had water polluted. Were performed twenty sample in each ponds and the following parameters assessed: pH, temperature, dissolved oxygen, turbidity, electrical conductivity, coliform thermotolerant, ammonia, organic nitrogen, TKN, nitrate, total phosphorus and alkalinity, bicarbonate, chloride, total hardness, calcium and magnesium hardness, total solids, TSS, TDS, COD and SAR°. To utilization on pisciculture we recommend use in ponds which presents the best quality. Only one pond presented higher pollutants and it not being appropriated for use in cooling tower. The content of solids suspension restricted the water use in all ponds in boilers. As water use in soil compaction, cleaning urban road and fire fighting system the water from these ponds are not appropriated. However, the recommended limits from literature are to domestic sewage, due to this, the parameters are restrictive to diminish the sanitary risk that could be offered with it utilization. The infiltration velocity of water these assessed ponds restrict a moderato use, however the effects are potentiated only in soil less sandy, which not occur in Natal. It is recommend the unrestricted irrigation utilization in ponds with best quality of water, and a restricted irrigation to ponds with worst quality of water. As load of pollutants, it is recommended a complementary treatment in those ponds whit higher load to diminish sanitary risks. In only one pond was found one helminth eggs and due to the convention formula of results, it was found for all ponds less than 1 helminth/liter eggs.

The influence of natural rubber/Au nanoparticle membranes on the physiology of Leishmania brasiliensis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Altered expression of the costimulatory molecules CD80, CD86, CD152, PD-1 and ICOS on T-cells from paracoccidioidomycosis patients: Lack of correlation with T-cell hyporesponsiveness

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Susceptibility to Yersinia pseudotuberculosis Infection is Linked to the Pattern of Macrophage Activation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)