Circulation of Eastern equine encephalitis, Western equine encephalitis, Ilhéus, Maguari and Tacaiuma viruses in equines of the Brazilian Pantanal, South America

Neutralizing antibodies to EEE (6.7%), WEE (1.2%), ILH (26.6%), MAG (28.2%) and TCM (15.7%) viruses were found in sera of 432 equines of the Brazilian Pantanal, area where undiagnosed horse deaths are frequently observed. A 4-fold rise in CF titer to EEE virus was detected in acute and convalescent sera of an encephalitis horse sacrified in 1992. Antibodies to EEE, ILH, MAG and TCM viruses were detected in horses less than 2 years old indicating recent circulation of these viruses in the Pantanal. The evidence of recent equine encephalitis associated with rising CF titer to EEE warrants a more intensive study with attempts to isolate virus from horses with clinical manifestations of encephalitis.

Standardization of an enzyme linked immunosorbent assay (ELISA) for detecting circulating toxic venom antigens in patients stung by the scorpion Tityus serrulatus

The sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA) for the detection of circulating antigens from toxic components of Tityus serrulatus scorpion venom was determined in patients stung by T. serrulatus before antivenom administration. Thirty-seven patients were classified as mild cases and 19 as moderate or severe cases. The control absorbance in the venom assay was provided by serum samples from 100 individuals of same socioeconomic group and geographical area who had never been stung by scorpions or treated with horse antisera. The negative cutoff value (mean + 2 SD) corresponded to a venom concentration of 4.8 ng/ml. Three out of the 100 normal sera were positive, resulting in a specificity of 97%. The sensitivity of the ELISA when all cases of scorpion sting were included was 39.3%. When mild cases were excluded, the sensitivity increased to 94.7%. This study showed that this ELISA can be used for the detection of circulating venom toxic antigens in patients with systemic manifestations following. T. serrulatus sting but cannot be used for clinical studies in mild cases of envenoming since the test does not discriminate mild cases from control patients.

Epidemiological aspects of the Brazilian spotted fever: serological survey of dogs and horses in an endemic area in the State of São Paulo, Brazil

In order to obtain information on Brazilian spotted fever, a study in domestic animals was performed in the County of Pedreira, State of São Paulo, Brazil, where 17 human cases had been notified. Serum samples obtained from animals were tested by indirect immunofluorescence for detectable antibodies to spotted fever-group rickettsiae. Seropositivity was revealed in 12 (36.4%) of 33 dogs and seven (77.8%) of nine horses from the endemic area. For comparison, blood samples from dogs and horses from non endemic area were tested and four (12.9%) of 31 dogs and three (27.3%) of 11 horses were positive. The highest titers of antibodies by IFA (IgG > 1:1024) were found only in three dogs and six horses from endemic area. The results suggest that dogs as horses may serve as environmental sentinels for estabilishing the prevalence of foci of spotted fever in Brazil.

Preliminary investigation of Culicidae species in South Pantanal, Brazil and their potential importance in arbovirus transmission

In view of the high circulation of migratory birds and the environmental and climatic conditions which favor the proliferation of arthropods, the Brazilian Pantanal is susceptible to circulation of arboviruses. However, the amount of data concerning arbovirus vectors in this area is scarce; therefore the aim of this study was to conduct a preliminary investigation of Culicidae species in the Nhecolândia Sub-region of South Pantanal, Brazil and their potential importance in the arbovirus transmission. A total of 3684 specimens of mosquitoes were captured, 1689 of which caught in the rainy season of 2007, were divided into 78 pools and submitted to viral isolation, Semi-Nested RT-PCR and Nested RT-PCR, with a view to identifying the most important arboviruses in Brazil. Simultaneously, 70 specimens of ticks found blood-feeding on horses were also submitted to the same virological assays. No virus was isolated and viral nucleic-acid detection by RT-PCR was also negative. Nevertheless, a total of 22 Culicidae species were identified, ten of which had previously been reported as vectors of important arboviruses. The diversity of species found blood-feeding on human and horse hosts together with the arboviruses circulation previously reported suggest that the Nhecolândia Sub-region of South Pantanal is an important area for arbovirus surveillance in Brazil.

IDENTIFICATION OF Pseudomonas spp. AS AMOEBA-RESISTANT MICROORGANISMS IN ISOLATES OF Acanthamoeba

Acanthamoeba is a “Trojan horse” of the microbial world. The aim of this study was to identify the presence of Pseudomonas as an amoeba-resistant microorganism in 12 isolates of Acanthamoeba. All isolates showed the genus Pseudomonas spp. as amoeba-resistant microorganisms. Thus, one can see that the Acanthamoeba isolates studied are hosts of Pseudomonas.

MOLECULAR INVESTIGATION OF HEMOTROPIC MYCOPLASMAS IN HUMAN BEINGS, DOGS AND HORSES IN A RURAL SETTLEMENT IN SOUTHERN BRAZIL

SUMMARY The aims of this study were to determine the prevalence of hemoplasmas in a rural Brazilian settlement's population of human beings, their dogs and horses, highly exposed to tick bites; to identify the tick species parasitizing dogs and horses, and analyze factors associated with their infection. Blood samples from 132 dogs, 16 horses and 100 humans were screened using a pan-hemoplasma SYBR green real-time PCR assay followed by a species-specific TaqMan real-time PCR. A total of 59/132 (44.7%) dog samples were positive for hemoplasmas (21 Mycoplasma haemocanisalone, 12 ' Candidatus Mycoplasma haematoparvum' alone and 21 both). Only 1/100 (1.0%) human sample was positive by qPCR SYBR green, with no successful amplification of 16S rRNA or 23 rRNA genes despite multiple attempts. All horse samples were negative. Dogs >1 year of age were more likely to be positive for hemoplasmas ( p= 0.0014). In conclusion, although canine hemoplasma infection was highly prevalent, cross-species hemoplasma transmission was not observed, and therefore may not frequently occur despite overexposure of agents and vectors.

Blood feeding patterns of Nyssomyia intermedia and Nyssomyia neivai (Diptera, Psychodidae) in a cutaneous leishmaniasis endemic area of the Ribeira Valley, State of São Paulo, Brazil

Introduction The aim of this study was to identify the blood feeding sources of Nyssomyia intermedia (Ny. intermedia) and Nyssomyia neivai (Ny. neivai), which are Leishmania vectors and the predominant sandfly species in the Ribeira Valley, State of São Paulo, Brazil, an endemic area for cutaneous leishmaniasis. Methods Specimens were captured monthly between February 2001 and December 2003 on a smallholding and a small farm situated in the Serra district in the Iporanga municipality. The blood meals of 988 engorged females were tested using the avidin-biotin immunoenzymatic enzyme-linked immunosorbent assay (ELISA). Seven blood meal sources were investigated: human, dog, chicken, bovine, pig, horse and rat. Results The results showed that among the females that fed on one or more blood sources, the respective percentages for Ny. intermedia and Ny. neivai, respectively, were as follows: human (23% and 36.8%), pig (47.4% and 26.4%), chicken (25.7% and 36.8%) and dog (3.9% and 0%), and the differences in the blood sources between the two species were statistically significant (p = 0.043). Conclusions Both species had predominant reactivity for one or two blood sources, and few showed reactivity indicating three or four sources. Many different combinations were observed among the females that showed reactivity for more than one source, which indicated their opportunistic habits and eclecticism regarding anthropic environmental conditions.

Eclectic feeding behavior of Lutzomyia (Nyssomyia) intermedia (Diptera, Psychodidae, Phlebotominae) in the transmission area of American cutaneous leishmaniasis, state of Paraná, Brazil

Introduction The blood meal source of sandflies provides valuable information about the vector/host interaction and allows for an understanding of American cutaneous leishmaniasis (ACL) transmission mechanisms. The aim of this study was to identify the blood meal sources of Lutzomyia (Nyssomyia) intermedia in an endemic area of leishmaniasis in Brazil's State of Paraná using a precipitin test. Methods Sandflies were collected in the rural locality of Epitácio Pessoa within the City of Adrianópolis, State of Paraná, in southern Brazil. A total of 864 female sandflies were captured, and 862 (99.8%) were identified as L. intermedia species. However, two unidentified specimens were considered to be part of the genus Lutzomyia. Results Among the females examined, 396 specimens presented reactions to a certain type of tested antiserum, and most (67.9%) reacted to the simple type. These sandflies fed mainly on the blood of birds, opossums, and rodents, but specimens that fed on the blood of humans, dogs, horses, cattle, and cats were also found. Among the cross-reactions found (32.1%), bird/rodent, bird/opossum, bird/dog, bird/human, and horse/dog cross-reactions were the most common. Conclusions These results demonstrate a tendency in the eclectic feeding behavior of L. intermedia and support its potential role as a vector for ACL in the study area.

Description of Dichelacera (Dichelacera) matogrossensis sp. nov. (Diptera, Tabanidae) of Brazilian Midwestern

ABSTRACT Dichelacera matogrossensis sp. nov. is described based on females from the Chapada dos Guimarães, state of Mato Grosso, Brazil. Diagnosis, discussion and illustrations are provided.

Ixodidas brasileiros e de alguns paizes limitrophes

The knowledge of the Ixodidae becomes every day, more and more important owing to the fact of the increasing number of diseases of man and animals they can transmit. In Brasil besides transmitting treponemosis, piroplasmosis and anaplasmosis to several domestic animals, the ticks are also responsible fo the transmission of the brazilian rocky mountain spotted fever (A. cajennense and Amblyomma striatum) and they can also harbour the virus of the yellow fever and even to transmit it in laboratory experiments (A. cajennense, O. rostratus). The Brazilian fauna of ticks is a small one and has no more than 45 well-established species belonging to the genus Argas, Ornithodoros, Ixodes, Haemaphysalis, Rhipicephalus, Boophilus, Amblyomma and Spaelaeorhynchus. The genus Amblyomma is the best represented one, with 67% of all species of ticks known in Brazil. One of the most important species in the Amblyomma cajennense owing to its abundance and its wide parasitism in many vertebrates: reptiles, birds and mammals, incluing man, who is much attacked by the larva, the nymph and the adult of this species. The other ticks who attack the man are the Amblyomma brasiliense (the pecari tick), in the forests, and the Ornithodoros, especially the species. O. rostratus and brasiliensis. Other species can bite the man, but only occasionally, like Amblyomma fossum, striatum, oblongogutatum etc. Argas persicus, Rhipicephalus sanguineus and Boophilus are very important species not only as parasites but specially because they transmit several diseases to animals. Some of the ticks of the brazilian wild animals are now also parasites of the domestic ones and vice-versa. Arga persicus var. dissimilis is very common among the poultry and transmits the Treponema anserinum (gallinarum). Boophilus microplus is very abundant on our domestic and wild ruminants (Bos, Cervus, Mazama etc.) and can also ben found on horse, dogs, Felis onca, Felis concolor etc., and it transmits to cattle piroplasmosis and anaplasmosis. Rhipicephalus sanguineus (an introduced species) is now very common on the dog, over all the country. The author recommend to give popular names to some brazilian ticks in order to make them more acquainted with the non scientific people. The author gives a classification of the superfamilia Ixoidoidea and keys to the determination of the different species of brazilian ticks. He creates a new family of Nuttallielidae to the so interesting tick, described by Bedford with the name of Nuttaliella namaqua in South Africa, a new variety of Argas persicus, the Argas persicus var. dissimilis nov. var. owing to the differences on the segment and on the size and morphology of the peritrema. He describes also the female of Amblyomma fuscum Nn. A great part of the author's work deals with the biology, life conditions and parasitism of many of the brazilian ticks in accordance with his personal and from other author's researches, especially in reference to Argas persicus, Ornithodoros rostratus, O. brasiliensis, Boophilus microplus, Rhipicephalus sanguineus, Amblyomma cajennense, A. pseudoconcolor, A. auriculare, A. rotundatum (= A. agamum) etc. The author gives a detailed report upon the parthenogenesis of A. rotundatum (A. agamum) that he first described in 1912 and gives also many references to other species of brazilian ticks, to teratological forms etc. He also gives a detailed report of the geographical distribution of brazilian ticks and of the peculiar conditions of its parasitism. The last part of this article deals with references to the species of ticks of some of the South American Republics namely Argentina, Bolivia, Colombia, Paraguay and Venezuela. Amblyomma testudinis Conil, A. neumanni Ribaga 1902 (= A. furcula Dõnitz 1909) and A. parvitarsum Nn. 1899 (= A. altiplanum Dios 1917), are found only in Argentina. It is given a special bibliography dealing with the brazilian ticks and four text figures and one plate.

Observaçães sôbre a congelação do plasma humano para desidratação

A technics for prefreezing of blood plasma and serum is described in this paper. The method indicated by Strumia et al. (2), uses a rapid local freezing to obtain the shell-freezing, with refigerated alcohol bath, at temperatures around minus 35ºC. On our work, it has been found that normal horse blood plasma fulfils the instructions given by Strumia, although normal human blood plasma, very often, fails to give the expected results. This is very disadvantageous at the routine work. With the use of small amounts of solid carbon dioxide, spread over the flasks, in the refrigerated bath, it has been possible to start the chrystallization. The technics prescribes a rapid cooling, like the one used by Strumia, to bring the temperature down, to about plus 10ºC. and, with rotating device stopped, the solid carbon dioxide is applied for one minute simultaneously on each flask. Starting rotation again, it begins to form a very uniform shell around the walls of the flasks.

Excreção urinária de 17-Cetoesteroides neutros no cavalo normal e no cavalo castrado

Total urinary neutral 17-steroids were determined in normal and in castrated horses. One liter of a 15-26 hours urine collection was hydrolysed by refluxing with 10% HC1 (v/v) for ten minutes and extracted with peroxyde-free ethyl ether. The extract was purified by washing with saturated NaHCO³ and KOH solutions. One half of the crude neutral fraction was fractionated with Girard's "T" reagent . The Zimmermann reaction was performed both in the ketonic and in the crude neutral extracts, using alcoholic 2.5N KOH and a 60 minutes period for the colour development in the dark. Optical density measuments were made in a grating Coleman Universal Spectrophotometer at 420 mµ and 520mµ; for the crude neutral fraction a colour correction equation was applied. The aliquot fraction used for colorimety was adjusted for keeping optical density measurements within the range 0.2 to 0.7. Androsterone (mp. 184-184.5°C) with an absorption maximum at 290.5 mµ (Beckman Model DU Spectrophotometer) was used as a reference standard. Table I, ilustrates the results obtained. At the 0.05 probability level there is a significant difference among castrated and normal group means (Fischer's "t" test.) when were used the data obtained from the ketonic fractions; in spite of the use of a colour correction applied for inespecific chromogens, the same results could not be obtained with the crude neutral fractions, Since Girard's reagent fractionation is generaly accepted as the best method for correcting the inespecific chromogen interference in the determination of the 17-ketosteroids by the Zimmermann reaction, we emphasize the value of the results obtained with the ketonic fractions. From these results it appears, as occurs in others mammals, that castrated horses show a lower level of urinary 17-ketosteroids excretion than the normal horses. The significance of the horse testis contribution for the neutral urinary steroid metabolites is discussed. Since horse urine has a low androgenic activity, the fractionation of the neutral 17-ketosteroids must be studied more accurately.

The ribonucleic acid content of some mammalian erythrocytes

RNA was determined in red blood cells of man and other mammals. Our report is based on 41 determinations. Red blood cells of rat showed the highest values in comparison with the blood cells of guinea pig, rabbit, horse and sheep which showed the lowest values, and man with intermediate ones. The method used was a combination of Schimidt and Thanhauser and Schneider extractions with the final reactions of pentose with the orcinol reagent colorimetrically measured.

Notas sôbre o gênero Colobogaster Solier, 1833, com a descrição de duas novas espécies da Região Amazônica: (Coleoptera, Buprestidae)

The study of materials belonging to several brazilian collections led us discover 2 new species of the genus Colobogaster which are here described. C. seabrai sp. n. seems to be related to C. puncticollis Waterhouse, 1882, from which it can be distinguished by: a) apical alitral tooth placed suturally, b) pronotum with 3 pairs of depressions, the 1st. pair transversal and conigous to the 2nd one, c) elitral suture brilliantgreen coloured but not the marginal edge, d) front without a horse-shoe shaped structure, e) pronotum with the discal region concolor. The structures of pronotum, the elitral and pronotal colour paterns and the genial morphology separate this one from other species of the genus. C. paraensis sp. n. is closely related to C. cupricollis Kerremans, 1897, but it is distinguished by the absence of depressions on the pronotum, by the elitral tooth placed suturally, by the abscence of humeral rip and by the general colour. Eleven other species were studied and their apical segment of the abdomen and scutellum were illustrated. It was also established the synonymy of C. ecuadoricus Obengerger, 1926.

The neotype of anopheles albitarsis (Diptera: culicidae)

Anopheles albitarsis neotype is described from specimens collected in Baradero, Argentina, in Shannon's trap, in horse and pig stables and on the progeny of engorded females. The description includes illustrations of adult female, male and female genitalias, scanning electron miscroscopy of the eggs and complete chaetotaxy of pupa and larva. The importance for electing a neotype is based on the realization that An. albitarsis is a complex of cryptic species. It is an attempt to provide typt-locality specimens with which other memebers of the group can be compared.