Mutagenic and antimutagenic effects of aqueous extract of rosemary (Rosmarinus officinalis L.) on meristematic cells of Allium cepa.

Polyphenolic compounds present in rosemary were found to have antioxidant properties, anticarcinogenic activity, and to increase the detoxification of pro-carcinogens. The aim of the study was to determine the effect the aqueous extract of rosemary (AER) on mutagenicity induced by methylmethane sulfonate in meristematic cells of Allium cepa, as well as to describe its mode of action. Anti-mutagenicity experiments were carried out with 3 different concentrations of AER, which alone showed no mutagenic effects. In antimutagenicity experiments, AER showed chemopreventive activity in cultured meristematic cells of A. cepa against exposure to methylmethane sulfonate. Additionally, post-treatment and simultaneous treatment using pre-incubation protocols were the most effective. Evaluation of different protocols and the percent reduction in DNA indicated bioantimutagenic as well desmutagenic modes of action for AER. AER may be chemopreventive and antimutagenic.

Glyphosate adsorption on C60 fullerene in aqueous medium for water reservoir depollution.

The indiscriminate use of pesticides has caused several damages to the environment, in particular the pollution of water reservoirs, so that this has motivated the development of techniques to minimize its consequences. One of the main surface water pollutants is glyphosate, which is a widely used herbicide for weed control. Therefore, in this work, computational simulations were used with density functional theory and molecular dynamics to theoretically verify if C60 fullerene is capable of adsorbing glyphosate in aqueous media. As a result, we showed through the adsorption energies, molecular dynamics methods, and infrared absorption that C60 can adsorb glyphosate molecules in at least three distinct configurations, either in vacuum or in water, which theoretically indicates it as a good candidate for removal of this herbicide from water by nanotechnology techniques.

Morphology and development of Anaplasma marginale in midgut of engorged female ticks of Boophilus microplus.

Morphology and development of Anaplasma marginale were studied in midgut epithelial cells of adult Boophilus microplus females using light and transmission electron microscopy. Hemoparasite-free B. microplus larvae were used to infest Holstein calves experimentally inoculated with A. marginale. When A. marginale parasitemia varied from 0.4% to 4.3%, 140 engorged female ticks were collected and kept individually in steel wire tubes and incubated outdoors on Brachiaria decumbens grass pasture. Female ticks in groups of 20 were dissected 14, 19, 26, 33, 40, 47 and 50 days after detachment. Engorged female ticks collected from an A. marginale-free calf were kept under the same conditions as control ticks. Gut sections of infected and control ticks were processed using light and electron microscopy. Infected gut sections were processed by the immunocytochemical peroxidase-antiperoxidase technique. Under light microscopy A. marginale colonies were found in midgut epithelial cells of engorged B. microplus females from 19 days on after detachment from donor calves. Colonies were 6.1-10.3 microns in diameter, ovoid or irregular and contained round or shaped organisms. Two distinct morphological types of colonies (BM1 and BM2) were found. BM1 colonies had a dense matrix strongly stained in which organisms were barely visible; BM2 had large separated organisms densely stained inside a matrix which was lightly stained. A. marginale colonies immunocytochemically stained were found in gut tissue sections using light microscopy. Using transmission electron microscopy organisms of different morphologies were found in midgut epithelial cells of engorged females, suggesting sequential phases of development of A. marginale in the vector. Modifications occurred in colonies and in the organisms within them. No symbiotic rickettsiae were found in intestinal cells of control ticks.

Eimeria caprina sp. n. from the domestic goat, Capra hircus, from the USA.

The ellipsoidal to slightly ovoid oocysts of Eimeria caprina sp. n. from the domestic goat, Capra hircus, are 27--40 x 19.5--26 micrometers (mean 31.8 x 23.1 micrometers); their sporocysts are 13--17 x 7--10 micrometers (mean 15.3 x 8.5 micrometers). The oocyst wall is 1.7 micrometers thick, smooth, dark brown to brownish-yellow, and 2-layered. Micropyle, polar granule, and sporocyst residuum are present; micropylar cap and oocyst residuum are absent. These features distinguish the new species from other species in the genus which have a micropylar cap, or are smaller, or have a small micropyle with a small internal plug.

Endogenous development of Eimeria minasensis in experimentally infected goats.

The endogenous development of Eimeria minasensis was studied in 9 coccidia-free goat kids inoculated with 10(5) sporulated oocysts/kg body weight. Kids were killed 4, 7 (2 animals), 10, 13, 16, 18, 19, and 22 days after inoculation (DAI). In tissue sections of the intestines stained with hematoxylin and eosin and examined by light microscopy, 2 generations of meronts, gamonts, gametes, and oocysts were found. The first generation of meronts developed in cells deep in the lamina propria of the jejunum and ileum. Mature giant meronts (299.4x243.8 microm) found 16 DAI were visible to the naked eye and contained a large number of crescent-shaped merozoites. The second generation of meronts developed in the epithelial cells of crypts of the ileum and above the host cell nuclei. Mature meronts (11.5x10.1 microm) with 18-28 comma-shaped merozoites were first seen 16 DAI. Gametogenesis took place in epithelial cells of the crypts and villi of the terminal part of the ileum, cecum, and colon. Macrogametes (27.8x17.6 microm), mature microgamonts (21.3x17.0 microm), microgametes, and oocysts (30.5x19.4 microm) were found 19 DAI. Sexual stages were below the host cell nucleus.

Development of Eimeria ninakohlyakimovae Yakimoff & Rastegaieff, 1930 emend. Levine, 1961 in experimentally infected goats (Capra hircus).

The endogenous development and prepatent and patent periods of Eimeria ninakohlyakimovae were studied in 43 1-3-wk-old coccidia-free kids inoculated with 5.0 x 10(4), 1.5 x 10(5), 2.0 x 10(5), or 9.0 x 10(5) sporulated oocysts/kg. Twenty-five kids were killed at 24- or 48-hr intervals, 2-18 days after inoculation (DAI). Two generations of meronts, gamonts, gametes, and oocysts were found in sections stained with hematoxylin and eosin and examined using under light microscopy. The first generation of meronts developed in the endothelium of the lacteals, in the lamina propria, and in the lymphatic vessels of the ileum submucosa. Mature, first-generation meronts, 165.5 x 123.6 microm, were first found 10 DAI. Second-generation merogony developed in the crypt epithelial cells of the cecum and colon; mature meronts, 16.8 x 11.6 microm, were first seen 12 DAI. Gametogenesis occurred in the cecum and colon epithelium; mature microgamonts (16.1 x 13.0 microm), microgametes, macrogametes (14.7 x 12.5 microm), and oocysts (18.3 x 13.3 microm) were seen at 13 DAI. The course of the infection was followed in 18 kids examined every day until 24 DAI. The prepatent period was 14.7 (13-17) days and the patent period 6.8 (4-10) days. The sporulation time at 30 C, with constant aeration, was 2-3 days.

Experimental infection of bovines with oocysts of Toxoplasma gondii.

Calves aged 3 months were readily infected with oocysts and cysts of Toxoplasma gondii administered by oral route. Fever, respiratory distress, nasal discharge and hyperemia of the conjunctivas were the most significant clinical signs noted in the infected animals. Parasitemia was demonstrated in all infected calves. It occurred on different days and up to 62 days after the infection. Toxoplasma was demonstrated in tissues of all infected calves, and the organ most frequently parasitized was the lymph node. Parasitism of the retina was demonstrated in two calves. All infected animals had antibody against T. gondii in their serum. The Sabin-Feldman dye test and the indirect immunofluorescent test were both useful in detecting antitoxoplasma antibody.

Besnoitia sp. (Protozoa:Toxoplasmatinae) from Akodon montensis (Rodentia:Cricetidae) in Santa Catarina State, Brazil.

Tissue cysts of Besnoitia sp. were found in muscles and several organs from a naturally infected Akodon montensis captured in the rural area of the municipality of Timbó, Santa Catarina State, in southern Brazil. Indirect fluorescence and enzyme-linked immunosorbent assays carried out with sera from mice chronically infected with Toxoplasma gondii and Besnoitia sp. showed, as expected, a stronger reaction against homologous than heterologous antigens. No cross-protection was observed in mice immunized with T. gondii when challenged with Besnoitia sp. This is the first description of a natural infection of A. montensis by parasites of the genus Besnoitia sp. in Brazil.

Outbreak of cryptosporidiosis in dairy goats in Brazil.

An outbreak of cryptosporidiosis is reported among 22 suckling kids aged one to two weeks which were maintained for experimental purposes at the veterinary hospital of the University of Minas Gerais. They were divided into three groups. Group A consisted of 10 animals with acute diarrhoea; initially their faeces were pasty but later they were excreted in watery streams. The animals were treated with gentamicin and fluid therapy but did not respond to treatment and died within a week. Postmortem examination revealed liquid intestinal contents, enlarged mesenteric lymph nodes, and hyperaemia and haemorrhage were observed in the final third of the small intestine of some of the animals. Of the 10 animals in group B four had diarrhoea and six were normal; postmortem examination showed that the macroscopic changes were similar to those observed in group A. Three of the animals with diarrhoea had a massive Cryptosporidium infection in the final third of the small intestine, caecum and colon. Four of the six normal animals had a moderate Cryptosporidium infection in the same organs. The two kids in group C died; they had pasty faeces, and there were many Cryptosporidium oocysts in the faecal smears.

Surgical repair of a pseudoaneurysm of the ascending aorta after aortic valve replacement.

We report the case of a patient with a pseudoaneurysm of the ascending aortic clinically diagnosed 5 months after surgical replacement of the aortic valve. Diagnosis was confirmed with the aid of two-dimensional echocardiography and helicoidal angiotomography. The corrective surgery, which consisted of a reinforced suture of the communication with the ascending aorta after opening and aspiration of the cavity of the pseudoaneurysm, was successfully performed through a complete sternotomy using extracorporeal circulation, femorofemoral cannulation, and moderate hypothermia, with no aortic clamping.

Enraizamento de estacas de genótipos de Camellia sinensis L. em meio ácido, presença de alumínio e ácido indolbutírico / Rooting of Camellia sinensis L. in acid medium, the presence of aluminum and indolebutyric acid

RESUMO Experimentos foram conduzidos para avaliar o efeito do meio ácido e do alumínio, assim como para determinar a concentração mais apropriada de ácido indolbutírico (AIB) para o enraizamento de estacas de diferentes genótipos de Camellia sinensis L (planta de chá). Para tal, foram coletados ramos de plantas-matrizes em Pariquera-Açu-SP, Brasil, no inverno de 2012 e preparadas estacas semi-lenhosas, contendo uma gema e uma folha, que foram mantidas em viveiro com 70% de sombreamento. A irrigação do substrato foi feita com água e soluções contendo ácido fosfórico e sulfato de alumínio a pH 5,5, 4,5; 3,5. No primeiro experimento, o delineamento experimental foi o inteiramente casualizado em esquema fatorial 3 x 7, três genótipos (F 15, IAC 259 e Comum) e sete condições diferentes de enraizamento (vermiculita a pH 6,5, vermiculita acidificada com ácido fosfórico a pH 3,5; 4,5 e 5,5 ou vermiculita acidificada com sulfato de alumínio a pH 3,5; 4,5 e 5,5). No segundo experimento, o tratamento que promoveu o maior enraizamento no primeiro experimento (vermiculita acidificada com sulfato de alumínio a pH 3,5) foi combinado ao tratamento com AIB. O delineamento experimental utilizado foi o inteiramente casualizado em esquema fatorial 3 x 6, sendo três genótipos e seis concentrações de AIB (0, 2000, 4000, 6000, 8000 e 1000 mg L-1). A vermiculita acidificada com sulfato de alumínio a pH 3,5, combinada a aplicação de 10000 mg L-1 de AIB por 30 segundos, foi o tratamento mais adequado para o enraizamento de estacas semi-lenhosas dos genótipos F15, IAC 259 e Comum.

ABSTRACT The experiments were carried out to evaluate the effect of medium acidic and aluminum, as well as determine the most suitable concentration of indolebutyric acid (IBA) for rooting cuttings of different genotypes of Camellia sinensis L (tea plant). Such, stems were collected from mother plants in Pariquera-Açu, São Paulo state, Brazil, in winter 2012 and prepared semi-hardwood cuttings, with one bud and one leaf, which were kept in a nursery with 70% of shading. Irrigation substrate was taken with water and solutions containing phosphoric acid and aluminum sulfate at pH 5.5, 4.5; 3,5. In the first experiment, the experimental design was completely randomized in a factorial scheme 3x7, three genotypes (F 15, IAC 259 and Comum) and seven different conditions of rooting (vermiculite at pH 6.5, acidified with phosphoric acid or aluminum sulfate vermiculite at pH 3.5, 4.5 and 5.5). In the second experiment, the treatment that promoted the highest rooting in the first experiment (vermiculite acidified with aluminum sulfate at pH 3.5) was combined with treatment with IBA The experimental design was completely randomized in a factorial scheme 3x6, three genotypes and six concentrations of IBA (0, 2.000, 4.000, 6.000, 8.000 and 10.000 mg L-1). Vermiculite acidified with aluminum sulphate to pH 3.5, combined application of 10.000 mg L-1 IBA for 30 seconds was the most appropriate treatment for cutting propagation of genotypes F15, IAC 259 and Comum. Vermiculite acidified with aluminum sulfate at pH 3.5, combined treatment with 10,000 mg L-1 IBA for 30 seconds, was the most suitable conditions for the rooting of cuttings Camellia sinensis L.

Eimeria caprovina sp. n. from the domestic goat, Capra hircus, from the USA.

Oocysts of Eimeria caprovina sp. n. from the domestic goat, Capra hircus, are ellipsoidal, subspherical or slightly ovoid, usually flattened at the micropylar end. They measure 29.7 (26-36) x 23.7 (23-28) micron. The sporocysts are elongate ovoids, measuring 14.3 (13.7) x 8.3 (8-9) micron, with Stieda bodies at the narrow ends. The oocyst wall is 1.6 micron thick, smooth, dark-brown to brownish-yellow, and 2-layered. A micropyle, 6.2 (4-10) micron in diameter, polar granule, and sporocyst residuum are present: micropylar cap and oocyst residuum are absent.

Life cycle of Eimeria christenseni Levine, Ivens & Fritz, 1962 form the domestic goat, Capra hircus L.

The endogenous development of Eimeria christenseni was studied in 10 two-to four-week-old kids inoculated with 10(6)-10(7) sporulated oocysts. They were killed at intervals from two to 26 days after inoculation, and their tissues were examined for endogenous stages of the coccidian by light microscopy. Such stages were found in the small intestine and mesenteric lymph nodes. In the sexual cycle, two generations of meronts were found. The first generation developed in endothelial cells of lacteals in the jejunum In the sexual cycle, two generations of meronts were found. The first generation developed in endothelial cells of lacteals in the jejunun and ileum and mesenteric lymph nodes, and mature meronts were first seen 14 days after inoculation. The second generation developed in epithelial cells of the glands of Lieberkuehn in the jejunum and ileum and in mesenteric lymph nodes, and its mature meronts were first seen by 16 days. Sexual stages were present mostly in epithelial cells of the tips and sides of the villi and less frequently in crypt cells of the jejunum and ileum. Mature macrogametes and microgamonts and oocysts were also first seen by 16 days. The prepatent period was 17(14-23) days; the patent ranged from 8 to more than 30 days. Sporulation time was 3-4 days at 30 degrees C. E. christenseni was found to be pathogenic, kids inoculated with 1-5 X 10(5) sporulated oocysts exhibited the following signs: severe diarrhea, anorexia, polydipsia, poor hair coat, and extreme weakness. They recovered about a month later, but their growth rates appeared to be lower than those of uninoculated animals kept under the same conditions. One kid died 20 days after inoculation with 10(7) oocysts.

Eimeria minasensis n. sp. (Apicomplexa: Eimeriidae) in the domestic goat Capra hircus, from Brazil.

Eimeria minasensis n. sp. is described in the domestic goat Capra hircus from Brazil. Oocysts ellipsoidal are 35 x 24.5 (32-37.7 x 20.9-27.9) microns. Sporocysts elongate-ellipsoid are 15.2 x 9 (12.3-18.4 x 7.8-10.2) microns, with a Stieda body at the narrow end. Oocyst wall smooth and bilayered; outer layer about 1.2 (0.8-1.6) microns and colorless; inner layer about 0.5 (0.4-0.8) micron and dark-brown. Micropyle, a mound-shaped micropylar cap 1.6 x 8.9 (0.8-2 x 7-10.2) easily dislodged; one or more oocyst polar granules present. Oocyst residuum absent. Sporocyst residuum present, composed of many scattered granules. Sporozoites elongate, lying lengthwise, "head to tail" in the sporocysts; one or two refractile globules are usually visible. Sporulation time was 120 hr at 27 degrees C, prepatent period, 19 to 20 days and patent period 15 to 25 days. Gamonts, gametes and oocysts present in cecum and colon. Prevalence was 12.8% (6/47) in goats from Minas Gerais, Brazil.

Sporogony and experimental transmission of Babesia equi by Boophilus microplus.

The development of Babesia equi in salivary glands of adult female Boophilus microplus was observed under a light microscope using semithin sections stained with toluidine blue. Engorged nymphs were obtained from splenectomized foals experimentally infected with B. equi. As adults, they were then fed on rabbits for 5 days and the salivary glands of manually collected individuals were removed at intervals of 24 h. Sporozoites were found in type III granular acini cells between the 2nd and 5th days following feeding on the rabbits. Sporoblasts and sporozoites were observed in the same or adjacent acini cells in all the glands examined. The formation of the sporozoites occurred following the multiple division of the sporoblasts through a process of radial budding from the periphery of bodies resulting from multiple fission. Sporozoites were detected in smears of adult males stained with Giemsa, between the 2nd and 5th days following feeding by the ticks. Adults of B. microplus, fed during the nymphal phase on foals with patent parasitemia, transmitted sporozoites of B. equi to a splenectomized foal. The role of B. microplus in the transmission and epidemiology of B. equi is discussed.

Development of an ELISA system for detection of anti-Anaplasma marginale antibodies in cattle in Brazil.

An ELISA test was developed for detecting antibodies against Anaplasma marginale in bovine sera. Four antigenic preparations were produced from infected red blood cells. Some aliquots of this preparation were stored at -70 degrees C with 30% DMSO in phosphate-buffered saline (PBS) and others were lysed with 0.9% NH4Cl and stored at -20 degrees C. Typical anaplasmal structures were seen by electron microscopy in the antigenic preparations containing the erythrocytes that had been stored with DMSO. The performance of the ELISA test was evaluated by testing 298 positive serum samples collected from immunized cattle, 39 negative serum samples collected from cattle imported from areas free of A. marginale and 50 samples collected from cattle naturally infected in the field. The test gave a specificity of 94.87% and a sensitivity of 100%.

Antimalarial activity of saccharidic polymers of dapsone and sulfadimethoxine.

With the purpose of obtaining pro-drugs of dapsone and sulfadimethoxine, those chemotherapeutic agents were attached through covalent bonding to starch polymeric dialdehyde (Sumstar-190). The antimalarial activity of the two resulting compounds - the dapsone saccharidic polymer (PS6) and the sulfadimethoxine saccharidic polymer (PS7) - in mice experimentally inoculated with Plasmodium berghei was significantly increased with this molecular modification. Mice infected with malaria and kept without treatment together with others which received different doses of PS6 and PS7 were also partially or totally cured, possibly due to the ingestion of excrements containing the parent chemotherapeutic agents.

Ultrastructure of endogenous stages of Eimeria ninakohlyakimovae Yakimoff & Rastegaieff, 1930 Emend. Levine, 1961 in experimentally infected goat.

The ultrastructure of endogenous stages of Eimeria ninakohlyakimovae was observed in epithelial cells of cecum and colon crypts from a goat experimentally infected with 2.0 x 10(5) oocysts/kg. The secondary meronts developed above the nucleus of the host cell. The nucleus first divides and merozoites then form on the surface of multinucleated meronts. Free merozoites in the parasitophorous vacuole present a conoid, double membrane, one pair of rhoptries, micronemes, micropore, anterior and posterior polar ring, a nucleus with a nucleolus and peripheral chromatin. The microgamonts are located below the nucleus of the host cell and contain several nuclei at the periphery of the parasite. The microgametes consist of a body, a nucleus, three flagella and mitochondria. The macrogamonts develop below the nucleus of the host cell and have a large nucleus with a prominent nucleolus. The macrogametes contain a nucleus, wall-forming bodies of type I and type II. The young oocysts present a wall containing two layers and a sporont.