Increased natural killer activity does not prevent progression of experimental kala-azar

Kala-azar is the visceral form of leishmaniasis and it is caused by intracellular parasites from the complex Leishmania donovani. Golden hamster (Mesocricetus auratus) infected with Leishmania donovani develop a disease very similar to human Kala-azar. There is conspicuous hipergammaglobulinaemia and their T cells do not respond to stimulation with parasite antigens. We used this experimental model to evaluate the natural killer (NK) activity during the initial phase of the disease. Outbred hamsters infected by intravenous route with 5.106 amastigotes of L. donovani 1S showed a concurrent increase in the spleen weight and in the spleen cell number. Using the single cell assay we detected a significant increase in the percentage of NK effector cells on the 4th day of infection. Imprints from spleen and liver showed at days 14 and 28 a significant increase in the parasite burden. These results show that the increased NK activity in the beginning of the infection was not able to restrain the progression of the disease in this experimental model.

Ticks (Acari: Ixodidae) associated with wild animals in the Pantanal region of Brazil

This paper describes the identification of ticks from wild animals of the Pantanal region in Brazil as part of a comprehensive study about established and emerging tick-host relationships and related pathological aspects. Eighty-one animals were captured (representing 13 species six orders), and ticks were found on 63 (78%). Tick species identified included Boophilus microplus (Canestrini), Amblyomma cajennense (F.), Amblyomma parvum Aragão, Amblyomma pseudoconcolor Aragão, Amblyomma scalpturatum Neumann, Amblyomma nodosum Neumann, Amblyomma ovale Koch, and Amblyomma tigrinum Koch. Dragging from grasslands yielded negative results compared with the high concentration of ticks that were collected from leaves in the forests.

The detection of Toxoplasma gondii by comparing cytology, histopathology, bioassay in mice, and the polymerase chain reaction (PCR)

The objective of this study was to compare the different methods of detecting Toxoplasma gondii in sheep tissue, tested serologically positive by the indirect immunofluorescent antibody test (IFAT). Brain, diaphragm, and blood samples were collected from 522 sheep slaughtered at the São Manuel abattoir, São Paulo State, Brazil. Brain and diaphragm samples from IFAT seropositive animals were digested by both trypsin and pepsin and then injected into mice. Part of the digested samples was used to prepare slides for Giemsa staining and in the polymerase chain reaction (PCR). Tissue fragments were fixed in formalin and examined using hematoxilin-eosin (HE). Forty of the sheep (7.7%) were IFAT positive. T. gondii was isolated in 23 (59.0%) of the 39 mice with pepsin-digested brain samples and in 27 (69.0%) of the 39 with trypsin-digested brain samples. Injection of diaphragm samples led to T. gondii isolation in 26 (66.7%) of the 39 pepsin-digested samples and 21 (53.8%) of the 39 trypsin-digested samples. Cytological and hystopathological examination of both brains and diaphragms was negative in all examined sheep. PCR was positive in 7 (17.9%) of the trypsin and 2 (5.1%) of the pepsin-digested samples, while 9 (23.1%) of the trypsin and 3 (7.7%) of the pepsin-digested samples showed T. gondii DNA. T. gondii isolation rate in mice (n = 34; 85.0%) was significantly higher than detection by PCR (n = 15; 37.5%). © 2001 Elsevier Science B.V.

Invasive enteritis by Strongyloides stercoralis presenting as acute abdominal distress under corticosteroid therapy.

Overwhelming helminthiasis is still a problem in endemic areas, especially in immunocompromised patients. We report a case of invasive intestinal strongyloidiasis that was clinically expressed as acute abdominal distress in a 73-year-old man from São Paulo who had been receiving methylprednisone, 20 mg/day, for one year for osteoarthritis. A surgical specimen from the ileum revealed invasive enteritis with severe infestation by Strongyloides stercoralis. The patient died of sepsis 6 days after surgery. The possibility of invasive strongyloidiasis should be considered in the differential diagnosis of acute abdominal distress in patients undergoing immunosuppressive therapy.

Ticks (Acari: Ixodidae) associated with domestic dogs in Franca region, São Paulo, Brazil

Ticks on 140 domestic dogs from both urban and rural areas of Franca region in São Paulo state were identified with 102 dogs from urban areas and 38 from rural areas. Of urban dogs, 27.5% were infested exclusively by Rhipicephalus sanguineus ticks. Of the rural dogs, 36.8% were infested with the following tick species: R. sanguineus, Boophilus microplus, Amblyomma ovale and A. cajennense. Mixed infestations included a dog hosting A. cajennense and A. ovale and another with B. microplus and R. sanguineus. The most intense infestations were detected on urban dogs. Hemolymph tests of these ticks performed to detect rickettsial or Borrelia bacteria yielded negative results.

Pelecitus helicinus Railliet & Henry, 1910 (Filarioidea, Dirofilariinae) and other nematode parasites of Brazilian birds

We report Pelecitus helicinus Railliet & Henry, 1910 from 13 species of birds of 2 orders and 7 families, collected from the states of São Paulo and Mato Grosso, Brazil. All 13 constitute new host records for this nematode. In addition, we report the first record of Aprocta golvani Diaz-Ungria, 1963 from Brazil and Monasa nigrifrons (Bucconidae), as well as a number of other nematode records from Neotropical birds.

Ticks (Acari: Ixodidae) on Wild Marsh-Deer (Blastocerus dichotomus) from Southeast Brazil: Infestations before and after Habitat Loss

The lake from Porto-Primavera hydroelectric power station inundated an area of 2,200 km2 at the border of São Paulo and Mato-Grosso do Sul States, Brazil. Infestations by ticks were evaluated on 135 marsh deer, Blastocerus dichotomus (Illiger), captured before and after inundation. Ticks were collected for identification, and infestation level of animals was assessed by scoring. Deer were divided into four groups according to capture location and temporal relation to the inundation. Groups 1, 2, and 3 were captured before inundation. Group 4 was captured after inundation. Four tick species were found: Amblyomma cajennense (F.), Amblyomma triste Koch, Anocentor nitens (Neumann), and Boophilus microplus (Canestrini). Groups 1, 2, 3, and 4 had 30, 45, 100, and 96%, respectively, of animals carrying B. microplus ticks. A. triste was observed on 16, 22, 22, and 88% of animals from groups 1,2,3, and 4, respectively. A. nitens and A. cajennense were observed only on group 4, on 32 and 16% of the animals, respectively. Groups 1 and 2 had only 4.8 and 6.1% of animals with high infestation levels, respectively, and no ticks on 46.8% and 45.5% of the animals, respectively. Conversely, groups 3 and 4 lacked noninfested animals and had high infestation levels on 77.8 and 50% of deer, respectively. Marsh area shrinkage was blamed for higher infestation levels on deer from groups 3 and 4. The widespread presence of A. triste on marsh deer, a Neotropical tick species, raises the possibility of a natural host-parasite relationship.

Antigens from Rhipicephalus sanguineus ticks elicit potent cell-mediated immune responses in resistant but not in susceptible animals

In the present study we compared the immunological reactions between Rhipicephalus sanguineus tick-infested susceptible (dogs and mice) and tick-resistant hosts (guinea pigs), elucidating some of the components of efficient protective responses against ticks. We found that T-cells from guinea pigs infested with adult ticks proliferate vigorously in the presence of concanavalin A (ConA), whereas ConA-induced cell proliferation of tick-infested mice and dogs was significantly decreased at 43.1 and 94.0%, respectively, compared to non-infested controls. Moreover, cells from mice and dogs submitted to one or three successive infestations did not exhibit a T-cell proliferative response to tick antigens, whilst cells from thrice tick-infested guinea pigs, when cultured with either a tick extract or tick saliva, displayed a significant increase in cell proliferation. Also, we evaluated the response of tick-infested mice to a cutaneous hypersensitivity test induced by a tick extract. Tick-infested mice developed a significant immediate reaction, whereby a 29.9% increase in the footpad thickness was observed. No delayed-type hypersensitivity (DTH) reaction was detected. Finally, the differential cell count at the tick attachment site in repeatedly infested mice exhibited a 6.6- and 4.1-fold increase in the percentage of eosinophils and neutrophils, respectively, compared to non-infested animals, while a decrease of 77.0-40.9 in the percentage of mononuclear cells was observed. The results of the cutaneous hypersensitivity test and the cellular counts at the tick feeding site for mice support the view that tick-infested mice develop an immune response to R. sanguineus ticks very similar to dogs, the natural host of this species of tick, but very different from guinea pigs (resistant host), which develop a DTH reaction in addition to a basophil and mononuclear cell infiltration at the tick-attachment site. In conclusion, saliva introduced during tick infestations reduces the ability of a susceptible animal host to respond to tick antigens that could stimulate a protective immune response. As a consequence, the animals present a lack of DTH response and disturbed cellular migration to tick feeding site, which can represent a deficient response against ticks. © 2003 Elsevier B.V. All rights reserved.

Two Digestive Trypsins Occur in Three Species of Neotropical Anophelines

Trypsin activity increases in the midgut of Anopheles aquasalis, Anopheles albitarsis, and Anopheles darlingi after a bloodmeal. The activity returns to basal levels at the time the blood is completely digested. Affinity chromatography, reversed-phase high performance liquid chromatography (HPLC), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were used to sequentially purify the mosquito trypsins found in the midguts at 24 h after feeding. Amino-terminal sequencing of the purified trypsins showed the occurrence of two distinct trypsins in the midgut of each of the mosquitoes studied. The sequences obtained are similar to those of the trypsins of other hematophagous insects.

Ehrlichiosis in anemic, thrombocytopenic, or tick-infested dogs from a hospital population in South Brazil

Ehrlichia canis has a worldwide distribution, but clinical manifestations may vary geographically. We selected 129 dogs to determine prevalence of ehrlichiosis in dogs with anemia, thrombocytopenia, or ticks presented to a Veterinary Teaching Hospital in South Brazil. Of the 129 dogs, 68 carried the brown dog tick (Rhipicephalus sanguineus), 61 had thrombocytopenia (platelet count <150,000/μl), and 19 had anemia (PCV < 22%). Twenty dogs fulfilled more than one inclusion criteria. Ehrlichiosis was diagnosed by positive amplification of ehrlichial DNA by PCR using primers ECC and ECB that amplify a sequence of the 16S rRNA gene. Presence of E. canis was confirmed by cleavage of the amplified DNA using endonucleases HaeIII and AvaI. Fourteen of 68 (21%) dogs with ticks had ehrlichiosis, whereas 12 of 61 (20%) dogs presented with thrombocytopenia and 4 of 19 (21%) anemic dogs had ehrlichiosis. Similar results were obtained in dogs with thrombocytopenia and anemia (one of eight positive) and in dogs with thrombocytopenia and ticks (two of seven positive). All four dogs with anemia and ticks, and the dog that fulfilled all inclusion criteria yield no amplification of ehrlichial DNA by PCR. Based on our results, one in each five dogs infested by the brown dog tick, with anemia or thrombocytopenia had ehrlichosis. Contrary to widespread believe, ehrlichiosis was not the main cause for thrombocytopenia in our region. © 2003 Elsevier B.V. All rights reserved.

Amblyomma cajennense ticks induce immediate hypersensitivity in horses and donkeys

Since host immune reaction to ticks interferes with tick-borne pathogen transmission, it is important to recognize naturally occurring tick-host immune relationships to better understand the epidemiology of such infectious diseases. Amblyomma cajennense is an important tick-borne disease vector in the Neotropical region and horses maintain it in domestic environments. In the present work intradermal testing of A. cajennense tick exposed horses and donkeys using crude tick antigens was used to evaluate the type of hypersensitivity induced by infestations. Animals sensitized by A. cajennense infestation displayed an immediate hypersensitivity reaction at the antigen inoculation site. Foals sensitized with experimental infestations and field sensitized horses presented the most intense reactions (40% of ear thickness increase). Field sensitized donkeys presented less intense reaction reaching no more than 22% of mean thickness increase. Control horses (non-sensitized) had the least intense reaction, with a peak of no more 12% of increase. The presence of a prominent immediate hypersensitivity in equids sensitized experimentally or by field infestations indicates that A. cajennense ticks induce in this host an immune response that is associated with IgE production and which is known to be inappropriate against intracellular pathogens. Differences observed between horses and donkeys are discussed.

Avaliação da eficiência dos métodos Direto, COPROTEST e Rugai no diagnóstico da estrongiloidíase

Strongyloidiasis, a relatively common parasitism in tropical and sub-tropical areas, is the result of the infection by the smaller nematode, the Strongyloides stercoralis. Humans can be infected by this parasite, which has in its vital cycle free-life forms of male and female individuals able to live in the ground, and with another step necessary parasitism in the intestinal wall. The diagnostic of the infection is routinely done by the microscopic observation of the larva in stool samples and the high sensibility of urn method over another one allows an trustable and efficient diagnostic. The efficiency of three methods (Direct, COPROTEST and Rugai) used in the Parasitology Sector of the NAC-LACAL in Araraquara (SP) to diagnosis the strongiloidiasis were evaluated. A number of 2346 samples of stool of patients from NAC-LACAL and Nestor Goulart Reis Hospital were analyzed in the period between August and December of 2002. The Rugai Method with an positivity index of 65 % was elected as the most efficient of thee ones.

Resistance of Santa Inês and Ile de France suckling lambs to gastrointestinal nematode infections.

A trial was carried out to determine the resistance to natural infection by gastrointestinal nematodes in 12 Santa Inês and nine Ile de France lambs before weaning. Faecal samples were obtained for faecal nematode egg counts (FEC). Blood samples were collected to determine packed cell volume (PCV), total plasma protein levels and peripheral eosinophil counts. Most Ile de France lambs (77.8%) were treated with an anthelmintic at 43 days of age, while 50% off Santa Inês lambs were treated at weaning, 57 days of age. The mean PCV values were normal in Santa Inês lambs, while in Ile de France lambs showed lower values reaching 22.3% at 43 days of age. The lowest mean plasma protein values were observed in Ile de France lambs (4.13 g/dl) at 43 days of age and in Santa Inês lambs (5.0 g/dl) at 57 days of age. Before weaning, Santa Inês lambs were susceptible to natural infections by gastrointestinal nematodes but with a greater capacity to stand the adverse effects of parasitism compared to Ile de France lambs.

Espécies do gênero Eimeria Schneider, 1875 (Apicomplexa: Eimeriidae) em caprinos leitros mantidos em sistema intensivo na região de São Jose do Rio Preto, estado de São Paulo, Brasil.

Were examined 58 dairy goats, 41 kids and 17 adults, Saanen and Alpine breeds, males and females, in intensive system, in the São José do Rio Preto region, São Paulo state, for detection and identification of Eimeria spp. oocysts. The 58 feces samples analyzed by the Centrifugal-flotation technique, were positive for, at least, one Eimeria species. The Eimeria species found in this research were: E. ninakohlyakimovae (77,6%), E. jolchijevi (72,4%), E. alijevi (63,8%), E. christenseni (63,8%), E. arloingi (62,1%), E. caprovina (56,9%), E. hirci (50,0%) and E. caprina (48,3%). It follows that the high rate of the positive animals and the high frequency of the Eimeria species among the animals demonstrated that the disease is common in dairy goats, kids and adults, in intensive system.