Caenorhabditis elegans as a model to screen plant extracts and compounds as natural anthelmintics for veterinary use

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Detection of IgG antibodies to Neospora caninum and Toxoplasma gondii in dogs examined in a veterinary hospital from Brazil

A total of 163 dogs with neuromuscular, respiratory and/or gastrointestinal disorders, was admitted at the Veterinary Hospital, Federal University of Uberlandia, Brazil, and submitted to serology for Toxoplasma gondii and Neospora caninum. Assays for T gondii included indirect haemagglutination (IHA), indirect fluorescent antibody (IFAT-Tg), immunoenzymatic (ELISA), and immunoblotting (IB-Tg). Assays for N, caninum included IFAT-Nc and immunoprecipitation (IP-Nc). Based on concordant results by three serological tests (IHA, IFAT-Tg and ELISA) for T gondii, and divergent results further confirmed by IB-Tg for reactivity to TgSAG1, the 163 sera were divided into two groups: 59 (36%) Tg-seropositive samples and 104 (64%) Tg-seronegative samples. Antibodies to Neospora were detected in 11 (6.7%) out of 163 analyzed dog sera, with 5 (3.1 %) samples reactive to both parasites (Tg+/Nc+), and 6 (3.7%) reactive only to Neospora (Tg-/Nc+). Antibodies only to T: gondii were found in 54 (33%) samples. Among the 11 Neospora-positive sera analyzed by IB-Tg, the five sera Tg+/Nc+ showed strong reactivity to Toxoplasma antigens, especially to TgSAG1 (p30). No reactivity was observed to TgSAG1 in the six samples Tg-/Nc+. By TP-Nc, two highly immunodominant antigens (29 and 35 kDa proteins) were recognized by all 11 IFAT-Nc positive sera. Our results suggest that the infection by N, caninum can be concomitantly present in dogs from this area, although less common, and therefore should be considered in the differential clinical diagnosis with T. gondii in dogs presenting neuromuscular, respiratory and/or gastrointestinal disorders. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Nenneguya garavelli n. sp and Myxobolus peculiaris n. sp (Myxozoa : Myxobolidae) in the gills of Cyphocharax nagelli (Osteichthyes : Curimatidae) from Rio do Peixe Reservoir, Sao Jose do Rio Pardo, São Paulo, Brazil

The present work describes myxozoans found in Cyphocharax nagelli (Characiformes: Curimatinae) commonly called saguiru collected from Rio do Peixe Reservoir, Sao Jose do Rio Pardo, São Paulo, Brazil. From a total of 38 examined fish, 24 were infected with Henneguya garavelli n. sp. (63% prevalence) and two with Myxobolus peculiaris n. sp. (5% prevalence) in the gills. Spores were studied by staining and fresh spores were observed by differential interference contrast optics. Henneguya garavelli n. sp. differs from Henneguya iheringi , Henneguya occulta, Henneguya cesarpintoi, Henneguya santae, Henneguya pisciforme, Henneguya amazonica, Henneguya striolata, Henneguya leporinicola and Henneguya chydadea in spore length and from Henneguya travassosi, Henneguya adherens, Henneguya malabarica, Henneguya piaractus and also Henneguya chydadea in polar capsule length and tail length. Myxobolus peculiaris n. sp. was very different when compared to other species of Myxobolus in its morphology and the biggest size of spore body. The authors present tables with comparative measurements of Brazilian myxozoan parasites. (c) 2006 Elsevier B.V. All rights reserved.

Neoechinorhynchus curemai (Acanthocephala : Neoechinorhynchidae) in Prochilodus lineatus (Osteichthyes : Prochilodontidae)from the Parana River, Brazil

This work evaluated, parasitic infections by Neoechinorhynchus curemai (Acanthocephala: Neoechinorhynchidae) in Prochilodus lineatus captured between August 2000 and August 2001 in the Parana River, Presidente Epitacio, São Paulo, Brazil. of 87 fishes examined, 59 were infected (25 males and 34 females). High mean intensities occurred in August 2000 (45.2, range 2-204), September 2000 (28.5, range 11-73), October 2000 (59.3, range 2-250) and February 2001 (27.3, range 3-73). There was no relationship of rainfall with mean intensity and prevalence. Males were more parasitized (P < 0.05) than females. This work contributes to the knowledge of helminth parasites of fish from a little studied region of the Parana River, showing diversity in their fauna when compared to other places in the same river. (c) 2005 Elsevier B.V. All rights reserved.

Ocorrência e tratamento de monogenoides em alevinos de pampo (Trachinotus carolinus) cultivados experimentalmente na região norte do estado de São Paulo

This work had as objective tests therapeutic treatments seeking the monogenea eradication in fingerlings Florida Pompano (Trachinotus carolinus) servants in cage net in the area of Ubatuba, State of São Paulo. The fingerlings presented an accentuated weigh loss and strong discoloration and through observation of scraped of gills under light microscope were identified a high amount of monogenea. Three treatments were tested in form of baths: T1 (fresh water for 5 minutes); T2 (formalin: 1: 1.000 for 20 minutes) and T3: (formalin: 1:4.000 for 30 minutes). The treatments were appraised through scraped of gills, mounted among you laminate and laminulas and observed to the light microscope. It was observed that in the tested conditions all of the treatments were efficient in the elimination of the monogenea without presenting lethality to the fish. The treatment is recommended T I (take a bath in fresh water for 5 minutes) for the facility in the application and for the absence of use of chemical products.

Anthelmintic efficacy of oral trichlorfon solution against ivermectin resistant nematode strains in cattle

Infected calves from two different rural estates in Brazil were studied to assess the anthelmintic efficacy of oral trichlorfon against naturally occurring ivermectin resistant parasitic nematode strains. In experiment 1, infected animals were from a region where ivermectin resistant populations of Haemoncus placei, Cooperia punctata,Cooperia spatulata and Trichuris discolor have recently been identified. Six calves with natural gastrointestinal nematode infections were treated with 48.5 mg/kg aqueous trichlorfon administered orally and six calves acted as a non-treated control group. In experiment 11 24 naturally infected calves were selected to enter one of four treatment groups, six animals each received: 48.5 mg/kg oral trichlorfon; 200 mu g/kg subcutaneous 1% ivermectin; 630 mu g/kg subcutaneous 3.15% ivermectin; or no treatment (control group). Gastrointestinal helminths were counted and identified post-mortem at 7 days (trichlorfon and 1% ivermectin treated and untreated animals) or 14 days (3.15% ivermectin treated and untreated animals) after administration of the test agents. Experiment I identified a high level efficacy for oral trichlorfon against four helminth species that have previously been shown to be ivermectin resistant in this geographical region: percentage efficacy was 99.82% against adult H. placei, 99.18% against C. punctata, 99.33% against C. spatulata, 81.06% against T. axei, 98.46% against Oesophagostomum radiatum and 100% against T. discolor. Trichlorfon also showed activity against the ivermectin (1% and 3.15%) resistant helminth species identified in experiment 11, attaining efficacy levels of 99.17% against H. placei, 98.46% against C punctata and 100.00% against T. discolor. These findings indicate that oral trichlorfon is an effective treatment option in the management of cattle infected with ivermectin resistant helminths. (C) 2009 Elsevier B.V. All rights reserved.

An investigation of Leishmania spp. in Didelphis spp. from urban and peri-urban areas in Bauru (São Paulo, Brazil)

Blood and bone marrow samples were taken from 112 Didelphis spp., collected between March 2005 and February 2006, from urban and peri-urban areas of Bauru, São Paulo State, Brazil, to evaluate the hypothesis that these animals might constitute a reservoir of Leishmania spp. Anti-Leishman ia ssp. antibodies were screened in the serum samples using an enzyme-linked immuno-sorbent assay (ELISA) and the polymerase chain reaction (PCR). PCR was performed on fragments of DNA samples from Leishmania spp. using primers 13A and 13B, and showed a positive outcome in 91.6% of the 112 samples tested. of the 107 samples analyzed by ELISA, 71 % were positive. Evidence of epidemiological risk factors such as a circulating parasite and freely moving vectors suggests that Didelphis spp. may participate in the transmission cycle of Leishmania spp. in Bauru. (c) 2007 Elsevier B.V. All rights reserved.

Experimental toxoplasmosis in pregnant bitches

Three Toxoplasma gondii free bitches (No, 3-5) were fed 15 000 sporulated T. gondii oocysts at 56, 40 and 32 day of gestation and the outcome of the pregnancy was monitored. Two of the three dogs infected during pregnancy showed evidence of congenital infection and one aborted. Two control bitches not fed oocysts delivered eight uninfected healthy pups. This study demonstrated that I gondii can be congenitally transmitted in dogs when bitches are infected during pregnancy. (C) 1999 Elsevier B.V. B.V. All rights reserved.

Systemic inflammatory response indicators in rabbits (Oryctolagus cuniculus) experimentally infected with sporulated oocysts of Eimeria stiedai (Apicomplexa: Eimeriidae)

Hemograms and acute-phase proteins in adult male New Zealand White rabbits that had been experimentally infected orally with sporulated oocysts of Eimeria stiedai were evaluated over a 28-day period. Fifty animals were used, divided into two groups: group A infected with 1 x 10(4) sporulated oocysts of E. stiedai and group B inoculated with distilled water. on the seventh day after infection, the infected animals presented anemia and leukocytosis with neutrophilia and monocytosis. Protein fractionation by means of electrophoresis identified 19 acute-phase proteins with molecular weights ranging from 24 to 238 kD. Ceruloplasmin, transferrin and haptoglobin showed high levels on the seventh day after infection, with gradual increases in their concentrations until the end of the experimental period. Thus, from the data of the present study, E. stiedai is considered to be a pyogenic etiological agent for which the infection level can be monitored through the leukocyte count and serum concentrations of ceruloplasmin, transferrin and haptoglobin, and these can be recommended as complementary tests.

First isolation and molecular characterization of Toxoplasma gondii from finishing pigs from São Paulo State, Brazil

Toxoplasma gondii infection is widely prevalent in humans in Brazil. Among the food animals, pigs are considered the most important meat source of T. gondii for infection in humans. In the present study, we report the first isolation of viable T. gondii from finishing pigs in Brazil. Antibodies to T. gondii were found in 49 (17%) of 286 pigs prior slaughter using the modified agglutination test (MAT) at a serum dilution of 1:25. Attempts were made to isolate T. gondii from 28 seropositive pigs. Samples of heart, brain, and tongue from each pig were pooled, digested in acid pepsin, and bioassayed in five mice per pig. Viable T. gondii was isolated from seven pigs; all isolates were lethal for mice. Restriction fragment length polymorphism on products of SAG2 locus amplified by PCR revealed that two isolates were Type I and five were Type III. The results indicate that phenotypically and genetically T. gondii isolates from pigs from Brazil are distinct from isolates of T gondii from pigs in the USA. (c) 2005 Elsevier B.V. All rights reserved.

Natural infection of gastrointestinal nematodes in long-nosed armadillos Dasypus novemcinctus Linnaeus, 1758 from Pantanal wetlands, Aquidauana sub-region, Mato Grosso do Sul State, with the description of Hadrostrongylus speciosum n. gen. et n. sp (Molineidae : Anoplostrongylinae)

This study evaluated the gastrointestinal helminth fauna of long-nosed armadillos, Dasypus novemcinctus, from the Pantanal wetlands, Aquidauana sub-region, Aquidauana County, Mato Grosso do Sul State, Brazil. Thirteen species of nematodes, comprising seven genera and four families, were recovered from their gastrointestinal tracts. The following descriptors of infection were determined: prevalence, variation of intensity, average intensity and abundance. Hadrostrongylus speciosum n. gen. et n. sp. is first described here. (c) 2006 Published by Elsevier B.V.

Observations on helminth infections of free-living and captive rheas (Rhea americana) in Brazil

The present work describes helminth infection of eight free-living and 12 captive rheas (Rhea americana) from, respectively, Pantanal of Mato Grosso do Sul State, and Jaboticabal, São Paulo State, Brazil. Captive birds were young and had a high mortality rate, while free-living birds were adult and apparently healthy. Infections were evaluated by post-mortem examination of internal organs and recovery of helminths using standard parasitological procedures. Seven species of nematodes (Sicarius uncinipenis, Torquatoides crotophaga, Deletrocephalus dimidiatus, D. cesarpintoi, Paradeletrocephalus minor, Capillaria venteli and Dicheilonema rheae) and two species of cestodes (Houttuynia struthionis and Chapmania tauricolis) were identified. P. minor, which inhabits the large intestine, was the most common helminth in free-living birds (63.9%). In captive rheas, a mean parasitic load of 173 helminths per host was found. The gizzard of these birds was the most parasitized organ and S. uncinipenis was most common (92.5%). Parasitism of free-living and captive birds and associated pathology are discussed. (c) 2005 Published by Elsevier B. V.

Abundância e sazonalidade de dípteros (Insecta) em granja aviária da região nordeste do Estado de São Paulo, Brasil

Modem production systems accommodate broody hens in high densities, leading to the accumulation of excrement under the cages. This substrate is excellent for the development of sinantropic flies. Thus, the accomplishment of surveys in these places becomes essential, in order to plan better strategies of control. The present work aimed at studying the entornofauna and the seasonality of the species of dipterous present in the Crisdan poultry house located in the Municipality of Sao Joao da Boa Vista, the State of São Paulo, Brazil. In the period of January of 2001 to December of 2002, 1,012,595 flies were captured using the "jug-trap". The species were identified: Drosophi-la repleta (Wollaston, 1858), Musca domestica (Linnaeus, 1758), Ophyra spp., Hennetria illucens (Linnaeus, 1758), Fannia canicularis (Linnaeus, 1761), Chrysomya megacephala (Fabricius, 1794), and Sepsidae. More frequently D. repleta and M. domestica had added 99.47% of the dipterous. Increased rainfall and the collection months influenced the sampling of dipterous (P < 0.05). Drosophila repleta was the most abundant species, representing 91% of all captured flies. However, this diptera did not develop at the surveyed site since immatures were not captured therein.

Chaunus ictericus (Spix, 1824) as paratenic host of the giant kidney worm Dioctophyme renale (Goeze, 1782) (Nematoda: Enoplida) in Sao Cristovao district, Tres Barras county, Santa Catarina state, Brazil

Dioctophyme renale larvae have been found in cysts in the gastric wall of 5.17% (3/58) Chaunus ictericus specimens from Sao Cristovao district, Tres Barras municipality, Santa Catarina state, Brazil. However, larvae of this nematode were not found in sympatric Chaunus schneideri. The larvae caused a mild granulomatous reaction. This is the first report of paratenic hosts for D. renale in Brazil, and probably is also the first in the Neotropical region. (C) 2009 Elsevier B.V. All rights reserved.

Antibody and blood leukocyte response in Rhipicephalus sanguineus (Latreille, 1806) tick-infested dogs and guinea pigs

The dog is considered to be the natural host of Rhipicephalus sanguineus and is unable to develop appreciable resistance even after repeated feedings. The guinea pig develops strong resistance after one infestation with adult ticks. Antibody (IgG) titres against tick salivary gland antigens (SGAs) and blood leukocyte numbers in dogs and guinea pigs undergoing experimental R. sanguineus tick infestations were measured to detect a possible correlation with susceptibility or resistance of hosts. Since infested dogs develop an immediate hypersensitivity reaction to R. sanguineus antigens, total and anti-R. sanguineus SGA IgE levels were also measured in this host species. IgG and IgE antibody levels were determined by an enzyme-linked immunosorbent assay (ELISA) along three consecutive infestations of both hosts. Most dogs and guinea pigs displayed low IgG levels against R. sanguineus SGAs, though marked differences in individual response were observed. Although dog's total serum IgE levels increased significantly after infestations, no change in the amount of anti-salivary gland IgE was detected. Total and differential blood cell counts were determined in dogs and guinea pigs during primary and secondary infestation. In dogs, a tertiary infestation and a subsequent higher infestation level were also evaluated. Infested dogs did not display any alteration in blood leukocyte counts throughout the experiment. Guinea pigs, on the other hand, developed a significant basophilia during primary infestation which increased further during secondary infestation. These data reveal similarities and differences in the reactions of resistant and non-resistant hosts to ticks. They contribute for the understanding of such host-parasite relationships and will hopefully aid in the development of immune control of ticks. (C) 2003 Elsevier B.V. All rights reserved.