Sanitary conditions of a colony of urban feral cats (Felis catus Linnaeus, 1758) in a zoological garden of Rio de Janeiro, Brazil

The colony of urban stray cats living in the Rio de Janeiro zoological garden was studied in order to develop a population and health control program. As many cats as possible were captured during two months (47 animals) and were classified according to gender, age, weight and coat markings. They were submitted to a general health evaluation, examined for the presence of ectoparasites and sent to a surgical neutering program. All animals had a blood sample drawn for CBC, platelet count, heartworm and retroviruses detection. Capillary blood smears were made for hemoparasites detection. Coat marking and colors were tabby (59.7%), followed by solid black (17%); torbie (10.6%); bicolor (10.6%) and harlequin (2.1%). The only ectoparasites found were fleas, which infested 28% of the animals. The hemoparasites found were Haemobartonella felis (38%) and piroplasmas that could not be differentiated between Cytauxzoon spp. and Babesia spp. (47%). No cat was found infected by Dirofilaria immitis or FeLV (Feline Leukemia Virus), although FIV (Feline Immunodeficiency Virus) antibodies could be detected (21%). There was no correlation between hemoparasites and FIV infections. The estimated total cat population (mark-recapture method) was 59; 68% female and 32% male, suggesting that a neutering program is in fact needed.

Optimization of random amplified polymorphic DNA techniques for use in genetic studies of Cuban triatominae

Random amplified polymorphic DNA (RAPD) technique is a simple and reliable method to detect DNA polymorphism. Several factors can affect the amplification profiles, thereby causing false bands and non-reproducibility of assay. In this study, we analyzed the effect of changing the concentration of primer, magnesium chloride, template DNA and Taq DNA polymerase with the objective of determining their optimum concentration for the standardization of RAPD technique for genetic studies of Cuban Triatominae. Reproducible amplification patterns were obtained using 5 pmoL of primer, 2.5 mM of MgCl2, 25 ng of template DNA and 2 U of Taq DNA polymerase in 25 µL of the reaction. A panel of five random primers was used to evaluate the genetic variability of T. flavida. Three of these (OPA-1, OPA-2 and OPA-4) generated reproducible and distinguishable fingerprinting patterns of Triatominae. Numerical analysis of 52 RAPD amplified bands generated for all five primers was carried out with unweighted pair group method analysis (UPGMA). Jaccard's Similarity Coefficient data were used to construct a dendrogram. Two groups could be distinguished by RAPD data and these groups coincided with geographic origin, i.e. the populations captured in areas from east and west of Guanahacabibes, Pinar del Río. T. flavida present low interpopulation variability that could result in greater susceptibility to pesticides in control programs. The RAPD protocol and the selected primers are useful for molecular characterization of Cuban Triatominae.

Sensitivity of an immunoenzymatic test for detection of ant-L. brasiliensis antibodies compared to other tests used for the diagnosis of American cutaneous leishmaniasis

The diagnosis of American cutaneous leishmaniasis (ACL) is frequently based on clinical and epidemiological data associated with the results of laboratory tests. Some laboratory methods are currently being applied for the diagnosis of ACL, among them the indirect immunofluorescence reaction (IIFR), the Montenegro skin test (MST), histopathological examination, and the polymerase chain reaction (PCR). The performance of these methods varies in a considerable proportion of patients. After the standardization of an immunoenzymatic test (ELISA) for the detection of IgG in the serum of patients with ACL using a crude Leishmania braziliensis antigen, the results obtained were compared to those of other tests routinely used for the diagnosis. The tests revealed the following sensitivity, when analyzed separately: 85% for ELISA IgG, 81% for PCR, 64.4% for MST, 58.1% for IIFR, and 34% for the presence of parasites in the biopsy. ELISA was positive in 75% of patients with ACL presenting a negative MST, in 84.8% of ACL patients with negative skin or mucous biopsies for the presence of the parasite, and in 100% of cases with a negative PCR. Thus, ELISA presented a higher sensitivity than the other tests and was useful as a complementary method for the diagnosis of ACL.

Optimization of the Sybr Green real time PCR for the detection of Human Herpes Virus type 6 (HHV-6)

HHV-6 is the etiological agent of Exanthem subitum which is considered the sixth most frequent disease in infancy. In immuno-compromised hosts, reactivation of latent HHV-6 infection may cause severe acute disease. We developed a Sybr Green Real Time PCR for HHV-6 and compared the results with nested conventional PCR. A 214 pb PCR derived fragment was cloned using pGEM-T easy from Promega system. Subsequently, serial dilutions were made in a pool of negative leucocytes from 10-6 ng/µL (equivalent to 2465.8 molecules/µL) to 10-9 (equivalent to 2.46 molecules/µL). Dilutions of the plasmid were amplified by Sybr Green Real Time PCR, using primers HHV3 (5' TTG TGC GGG TCC GTT CCC ATC ATA 3)'and HHV4 (5' TCG GGA TAG AAA AAC CTA ATC CCT 3') and by conventional nested PCR using primers HHV1 (outer): 5'CAA TGC TTT TCT AGC CGC CTC TTC 3'; HHV2 (outer): 5' ACA TCT ATA ATT TTA GAC GAT CCC 3'; HHV3 (inner) and HHV4 (inner) 3'. The detection threshold was determined by plasmid serial dilutions. Threshold for Sybr Green real time PCR was 24.6 molecules/µL and for the nested PCR was 2.46 molecules/µL. We chose the Real Time PCR for diagnosing and quantifying HHV-6 DNA from samples using the new Sybr Green chemistry due to its sensitivity and lower risk of contamination.

Geographical distribution and intra-domiciliary capture of sylvatic triatomines in La Convención province, Cusco, Peru

The geographical distribution and intra-domiciliary capture of sylvatic triatomines in three districts of the province of La Convención, Cusco, Peru are presented. In the district of Vilcabamba, eight adults of Rhodnius pictipes and five adults of Panstrongylus geniculatus were found. In the district of Ocobamba, 19 adults, 14 nymphs, and eggs of P. rufotuberculatus were found. In the district of Echarate, six adults and 10 nymphs of Eratyrus mucronatus, an adult of R. pictipes and P. geniculatus, and a nymph of P. rufotuberculatus were also found. The geographical distribution of E. mucronatus has extended to Cusco. This is the first report in Peru of household colonization by this triatomine.

Characterization of rabies virus isolated from a colony of Eptesicus furinalis bats in Brazil

Some bat species have adapted to the expanding human population by acquiring the ability to roost in urban buildings, increasing the exposure risk for people and domestic animals, and consequently, the likelihood of transmitting rabies. Three dead bats were found in the yard of a house in an urban area of Jundiaí city in the state of São Paulo in southeast Brazil. Two of the three bats tested positive for rabies, using Fluorescent Antibody and Mouse Inoculation techniques. A large colony of Eptesicus furinalis was found in the house's attic, and of the 119 bats captured, four more tested positive for rabies. The objectives of this study were to report the rabies diagnosis, characterize the isolated virus antigenically and genetically, and study the epidemiology of the colony.

Optimization of randomly amplified polymorphic DNA-polymerase chain reaction for molecular typing of Salmonella enterica serovar Typhi

Optimization of the RAPD reaction for characterizing Salmonella enterica serovar Typhi strains was studied in order to ensure the reproducibility and the discriminatory power of this technique. Eight Salmonella serovar Typhi strains isolated from various regions in Brazil were examined for the fragment patterns produced using different concentrations of DNA template, primer, MgCl2 and Taq DNA polymerase. Using two different low stringency thermal cycle profiles, the RAPD fingerprints obtained were compared. A set of sixteen primers was evaluated for their ability to produce a high number of distinct fragments. We found that variations associated to all of the tested parameters modified the fingerprinting patterns. For the strains of Salmonella enterica serovar Typhi used in this experiment, we have defined a set of conditions for RAPD-PCR reaction, which result in a simple, fast and reproducible typing method.

Comparison of capture methods for the diagnosis of adult anopheline populations from State of Mato Grosso, Brazil

INTRODUCTION: The present study compares human landing catches of primary malaria vectors with two alternative methods of capture: the Shannon trap and the Mosquito magnet. METHODS: This study used regression models to adjust capture data to a negative binominal distribution. RESULTS: Capture numbers and relative percentages obtained from the three methods vary strongly between species. The highest overall captures were obtained for Anopheles triannulatus with captures for the Shannon trap and the Mosquito magnet measuring more than 330% higher than captures obtained by human landings. For Anopheles darlingi, captures by the Shannon trap and the Mosquito magnet were about 14% and 26% of human landing catches, respectively. Another species with malaria transmission potential that was not sampled by human landing captures weascaptured by the Shannon trap and the Mosquito magnet (Anopheles oswaldoi). Both alternative sampling techniques can predict the human landing of Anopheles triannulatus, but without proportionality. Models for Anopheles darlingi counts, after totaling daily captures, are significant and proportional, but prediction models are more reliable when using the Shannon trap compared with the Mosquito magnet captures. CONCLUSIONS: These alternative capture methods can be partially recommended for the substitution of human landing captures or, at least, as complementary forms of monitoring for malarial mosquitoes.

Capture of Triatoma arthurneivai (Hemiptera: Reduviidae) using a new luminous trap in Southeast Brazil

INTRODUCTION: Triatomine bug captures in the wild are laborious and time-consuming. Some traps may assist in this task. We report a new trap design and the capture of a specimen of a triatomine rarely found. METHODS: Two luminous traps were installed in the city of Diamantina, State of Minas Gerais, and surveyed weekly for a year. RESULTS: A Triatoma arthurneivai female and other triatomine bugs were caught. CONCLUSIONS: A new trap design may be used in triatomine bugs field captures, mainly in low density areas. We assume the center of endemism of Triatoma arthurneivai is the Espinhaço Mountain range.

Hybrid capture II and PapilloCheck® tests for detection of anal high-risk human papillomavirus

Introduction This study evaluated the level of concordance between hybrid capture II (HCII) and PapilloCheck® for the detection of high-risk human papillomavirus (HPV) in anal samples. Methods Anal cell samples collected from 42 human immunodeficiency virus (HIV)+ patients were analyzed. Results Considering only the 13 high-risk HPV types that are detectable by both tests, HCII was positive for 52.3% of the samples, and PapilloCheck® was positive for 52.3%. The level of concordance was 80.9% (Kappa = 0.61). Conclusions Good concordance was observed between the tests for the detection of high-risk HPV.

Local diversity of tropical and temperature ant faunas (Hymenoptera, Formicidae)

Qualitative and quantitative collections of ants made in the region of Manaus, Brazil (evergreen tropical humid forest), and in western North Carolina, USA (deciduous temperate/wet forest), were undertaken to investigate. latitudinal patterns of ant diversity and community organization on regional and local scales. We have found to date 307 ant species in the Municipality of Manaus. Totals ranging from 134 to 270+ species have been reported in the literature for other tropical regions of less than 10,000km2. In contrast, temperate ant surveys generally report only SO to 150 species in similar or larger areas. Sampling at sardine baits set 10m apart on square grids, we found forest ecosystems near Manaus to be much richer and more diverse in ants than those sampled in North Carolina: 28 species vs. 5-10 species in 50 collections and 16 vs. 3 previously unrecorded species discovered with each doubling of sample size. Room's (1975a) results from climatically simllar Papua New Guinea forest agree closely with those from Manaus. We suggest that one important factor contributing to the increased diversity of tropical, omnivorous ants may be greater variety of nest sites available for specialization.

Effects of forest fragmentation on amazonian understory bird communities

SUMMARYData form an intensive mist-netting mark-recapture program in the central Amazon demostrate significant changes in the undesrtory avian community in isolate patches of 1 and 10 ha of terra firme forest. Following isolation, capture rates increase significantly as birds fleeing the felled forest entered the. newly formed forest fragments. Movement to and from the reserve is restricted, as witnessed by an increase in recapture percentages following isolation. Species of birds that axe. obligate army ant followers disappeared at the time the surrounding habitat was removed from 1 - and 10 - ha areas. The complex mixed-species insectivorous flocks typical of Amazonian forests deteriorated within 2 years of isolation of 1 - and 10 - ha forest fragments. Several species of mid-story insectivores changed their foraging behavior after isolation of small forest reserves.

The persistence of ant-following birds in small amazonian forest fragments

SUMMARYMist-net captures of army ant-following bird were monitored during the isolation of central Amazonian forest fragments of 1 ha (n = 5), 10 ha (n = 4) and 100 ha (n = 1). Post-isolation captures of the three obligate ant-following birds Dendrocinela merula, Pithys albifrons, and Gymnopithys rufigula decreased significantly in all fragments. post-isolation captures of four facultative ant-following species were not significantly different in forest fragments of 1 ha and 10 ha, although two species decreased significantly in the 100 ha fragment. Experimental introductions of obligate species into small forest fragments in the absence and presence of introduced Eciton burchelli army ant colonies resulted in significantly greater recaptures of introduced birds when active army ant colonies were present. Of the 105 birds introduced, 58 ' 55%) crossed 100-320 m od dedorested area an were recaptured in continuous forest.

Five egg-laying queens in a single colony of brazilian stingless bees (Melipona scutellaris Latreille

Polygyny, characterized by the presence of several egg-laying queens, is considered as a temporary colony status. In stingless bees it is rarely observed. This paper reports the first case of natural polygyny in Melipona scutellaris colony, with five egg-laying queens.