Dynamics of Ctenocephalides felis felis (Siphonaptera: Pulicidae) Infestations on Urban Dogs in Southeastern Brazil.

The cat flea, Ctenocephalides felis felis (Bouché, 1835), is an important ectoparasite of dogs and cats throughout the world, causing annoyance to the animals and acting as a vector of infections and a cause of allergic dermatitis in dogs and cats. Although climatic variability and seasonality are known to influence the diversity and abundance of fleas, few investigations of seasonal prevalence of cat flea infestation have involved the same group of dogs being examined regularly over an extended period. The aim of this study was to determine the effects of temperature, rainfall, and relative humidity on the infestation by C. felis felis on 88 outdoor dogs in southeastern Brazil. The dogs, which were of mixed breed, sex, and age, were examined for ectoparasites every month during the period August 2011 to July 2012, and samples of fleas were randomly collected and identified. Meteorological data, comprising mean temperature, total rainfall, and mean relative humidity, were recorded for the calendar month prior to that in which the examinations were performed. Dogs were found to be infested only with C. felis felis, with a higher prevalence in the months with lowest rainfall (July, August, and September). The data obtained in this investigation can be used in control programs in order to establish an efficient strategy for environmental management and the application of insecticides, particularly during the driest months of the year based on the seasonal pattern of infestation of dogs by C. felis felis.

A new species of Tunga (Siphonaptera: Tungidae) parasitizing cattle from Brazil.

Tunga hexalobulata (Siphonaptera: Tungidae), new species oftungid sand flea belonging to the penetrans group, is described with illustrations of adult female parasitizing Bos indicus (L., 1758) from Brazil. It differs from the 12 other known species of Tunginae by the presence of six anterior humps in the neosome. It also can be differentiated from other species of the penetrans group by lesser size of the neosome, presence of three posterodorsally bristles in antennal segment II, and the extension of the posterior arm of the preoral internal sclerotization.

A new species of Tunga (Siphonaptera: Tungidae) from Brazil with a key to the adult species and neosomes.

Tunga bossii new species of tungid sand flea belonging to the caecata group is described with illustrations of the adult female, parasitizing the wild rodent Delomys dorsalis (Hensel) from Brazilian Atlantic Forest. Tunga bossii differs from the ten other known species of Tunginae by the size of the first segment of the maxillary palp and the presence of two bristles at the base of the maxilla. Tunga bossii also can be differentiated from other species of the T. caecata group by the eye morphology. A key to the adult species and neosomes of the genus Tunga also is included.

Use of Multiple Displacement Amplification as Pre-polymerase Chain Reaction (Pre-PCR) to amplify genomic DNA of siphonapterids preserved for long periods in scientific collections.

The recently developed Multiple Displacement Amplification technique (MDA) allows for the production of a large quantity of high quality genomic DNA from low amounts of the original DNA. The goal of this study was to evaluate the performance of the MDA technique to amplify genomic DNA of siphonapterids that have been stored for long periods in 70% ethanol at room temperature. We subjected each DNA sample to two different methodologies: (1) amplification of mitochondrial 16S sequences without MDA; (2) amplification of 16S after MDA. All the samples obtained from these procedures were then sequenced. Only 4 samples (15.4%) subjected to method 1 showed amplification. In contrast, the application of MDA (method 2) improved the performance substantially, with 24 samples (92.3%) showing amplification, with significant difference. Interestingly, one of the samples successfully amplified with this method was originally collected in 1909. All of the sequenced samples displayed satisfactory results in quality evaluations (Phred ≥ 20) and good similarities, as identified with the BLASTn tool. Our results demonstrate that the use of MDA may be an effective tool in molecular studies involving specimens of fleas that have traditionally been considered inadequately preserved for such purposes.

Siphonaptera associated with wild mammals from the Central Atlantic Forest Biodiversity Corridor in southeastern Brazil.

We collected 379 samples of eight flea species from 235 specimens of wild mammals captured in three areas of the Central Atlantic Forest Corridor in the state of Espírito Santo, southeastern Brazil, between August 2006 and February 2008. Among the 22 mammal species, we found eight species of marsupials (order Didelphimorphia), 13 species of rodents (order Rodentia), and one species of wild cat (order Carnivora). The total prevalence of infestation was 28.1%, with similar percentages in rodents (26.4%) and marsupials (29.5%). The infestation in the southern area was 3 times higher than that observed in the northern area. The infestation by single flea species was > 6 times higher than the total number of double and triple infestations. The mean abundance of fleas was 1.61. The flea female/male ratio was 1.38, ranging from 0.95 in Adoratopsylla (Tritopsylla) intermedia intermedia (Wagner) to 1.81 in Hechtiella nitidus (Johnson) in the most flea species infested. We report three new records of flea species in the state of Espírito Santo, and eight new host records.

Seasonality and prevalence rates of Steinina sp. (Eugregarinorida: Actinocephalidae) in Ctenocephalides felis felis (Siphonaptera: Pulicidae) from dogs captured in Belo Horizonte, Minas Gerais, Brazil.

In total, 1500 specimens (448 males and 1052 females) of the flea Ctenocephalides felis felis (Bouché (Siphonaptera: Pulicidae) were collected over a period of a year from 150 dogs captured by the Centro de Controle de Zoonoses de Belo Horizonte, Minas Gerais, Brazil. Microscopic examination of the dissected fleas revealed that 180 fleas were infected with a species of gregarine that was subsequently identified as a member of the genus Steinina. The relative abundances, prevalence rates, and seasonal variation of the different developmental stages of this endoparasite in C. felis felis were determined. Both gamonts and gametocysts presented significant seasonal variation.

Especies de Amblyomma (acari: ixodidae) parasitando perros (canis familiaris) en áreas rurales de los estados Lara, Yaracuy, Carabobo y Falcón Venezuela / Species of Amblyomma (acari: ixodidae) parasitizing dogs (canis familiaris) from rural areas of Lara, Yaracuy, Carabobo y Falcón Venezuela

Un total de 50 caninos de áreas rurales de los estados Lara, Yaracuy, Carabobo y Falcón, Venezuela, fueron examinados visualmente y mediante palpación cutánea con el fin de detectar la presencia de garrapatas. Ninguno de los animales se encontró parasitado por Rhipicephalus sanguineus, la garrapata común del perro. Se colectaron 139 especimenes adultos del género Amblyomma Koch, 1844: 72 hembras y 65 machos de la especie Amblyomma ovale Koch, 1844, y dos hembras de la especie Amblyomma triste Koch, 1844. La identificación de los especimenes se realizó mediante observación en lupa estereoscópica y con la ayuda de claves taxonómicas. Los Estados donde se colectó el mayor número de ixódidos fueron Falcón con 52 especimenes y Lara con 49, representando 38 y 36%, respectivamente. A. ovale ha sido reportada como vector natural de Hepatozoon canis en áreas rurales. La especie A. triste fue colectada sólo en un canino del estado Yaracuy. Esta especie ha sido incriminada como el vector biológico de varios agentes rickettsiales patógenos, tanto para humanos como para animales domésticos y salvajes, por lo que se resalta la importancia del género Amblyomma en la transmisión de enfermedades zoonóticas.

Fifty dogs from the rural areas of Lara, Yaracuy, Carabobo and Falcón States of Venezuela were sampled through the visual and touch techniques in search of the presence of ticks. All the dogs were free of the brown dog tick Rhipicephalus sanguineus. A total of 139 adult ticks of the genus Amblyomma Koch, 1844 were collected: of them, 65 males and 72 females were identified as Amblyomma ovale Koch, 1844, whereas two females belonged to the species Amblyomma triste Koch, 1844. The observation through stereomicroscope, as well as the use of taxonomic keys, were employed in the identification of the collected specimens. Most of the ticks were collected in Falcón and Lara States, totalling 52 (38%) and 49 (36%), respectively. The species A. ovale has been reported as a natural vector of the protozoan Hepatozoon canis in rural areas. A. triste was collected only in one dog in Yaracuy State. This species has been incriminated as a biological vector of several rickettsial agents to the men and animals. The actual role of the genus Amblyomma as vector of zoonotic disease agents is highlighted.

Rickettsia infection in five areas of the state of São Paulo, Brazil.

This study investigated rickettsial infection in animals, humans, ticks, and fleas collected in five areas of the state of São Paulo. Eight flea species (Adoratopsylla antiquorum antiquorum, Ctenocephalides felis felis, Polygenis atopus, Polygenis rimatus, Polygenis roberti roberti, Polygenis tripus, Rhopalopsyllus lugubris, and Rhopalopsyllus lutzi lutzi), and five tick species (Amblyomma aureolatum, Amblyomma cajennense, Amblyomma dubitatum, Ixodes loricatus, and Rhipicephalus sanguineus) were collected from dogs, cats, and opossums. Rickettsia felis was the only rickettsia found infecting fleas, whereas Rickettsia bellii was the only agent infecting ticks, but no animal or human blood was shown to contain rickettsial DNA. Testing animal and human sera by indirect immunofluorescence assay against four rickettsia antigens (R. rickettsii, R. parkeri, R. felis, and R. bellii), some opossum, dog, horse, and human sera reacted to R. rickettsii with titers at least four-fold higher than to the other three rickettsial antigens. These sera were considered to have a predominant antibody response to R. rickettsii. Using the same criteria, opossum, dog, and horse sera showed predominant antibody response to R. parkeri or a very closely related genotype. Our serological results suggest that both R. rickettsii and R. parkeri infected animals and/or humans in the studied areas.

Rickettsia infection in five areas of the state of São Paulo, Brazil

This study investigated rickettsial infection in animals, humans, ticks, and fleas collected in five areas of the state of São Paulo. Eight flea species (Adoratopsylla antiquorum antiquorum, Ctenocephalides felis felis, Polygenis atopus, Polygenis rimatus, Polygenis roberti roberti, Polygenis tripus, Rhopalopsyllus lugubris, and Rhopalopsyllus lutzi lutzi), and five tick species (Amblyomma aureolatum, Amblyomma cajennense, Amblyomma dubitatum, Ixodes loricatus, and Rhipicephalus sanguineus) were collected from dogs, cats, and opossums. Rickettsia felis was the only rickettsia found infecting fleas, whereas Rickettsia bellii was the only agent infecting ticks, but no animal or human blood was shown to contain rickettsial DNA. Testing animal and human sera by indirect immunofluorescence assay against four rickettsia antigens (R. rickettsii, R. parkeri, R. felis, and R. bellii), some opossum, dog, horse, and human sera reacted to R. rickettsii with titers at least four-fold higher than to the other three rickettsial antigens. These sera were considered to have a predominant antibody response to R. rickettsii. Using the same criteria, opossum, dog, and horse sera showed predominant antibody response to R. parkeri or a very closely related genotype. Our serological results suggest that both R. rickettsii and R. parkeri infected animals and/or humans in the studied areas.

Investigations on the biology, epidemiology, pathology and control of Tunga penetrans in Brazil. VI. Natural history of the infestation in laboratory-raised Wistar rats.

Tungiasis is endemic in many countries in Latin America, the Caribbean and sub-Saharan Africa, and it is associated with severe morbidity. The pathophysiological and immunological characteristics of the ectoparasitosis are not well understood, and no effective therapy is currently available. The aim of this study was to describe the natural history of tungiasis in laboratory-raised Wistar rats. The rats were exposed in the laboratory to the parasite or were kept in a natural environment with an intense transmission of Tunga penetrans. The time course of the infestation was determined, and lesions were photographed, described clinically in detail and biopsied. Biopsies were examined histopathologically and by light and scanning electron microscopy. Based on these findings, the natural history of tungiasis in Wistar rats was described and divided in five stages. Our data show that the natural history of tungiasis in Wistar rats and humans is almost identical, except that in the animals, the basement membrane disrupts 5 days after penetration and provokes an intense infiltration of the dermis, while in humans, the basement membrane remains intact. The study indicates that the Wistar rat is an appropriate model for the study of clinical and pathological aspects of tungiasis. Using this model should enable a better understanding of the pathophysiology and immunology of the ectoparasitosis.

Endosymbionts of Ctenocephalides felis felis (Siphonaptera: Pulicidae) obtained from dogs captured in Belo Horizonte, Minas Gerais, Brazil.

Specimens of fleas Ctenocephalides felis felis (1052 female symbol/448 male symbol), obtained from 150 dogs in Centro de Controle de Zoonoses de Belo Horizonte, Minas Gerais, Brazil, were dissected and examined for endosymbionts. Three protozoan, Nolleria pulicis, a gregarine (Actinocephalidae) and Leptomonas sp., together with one cestode, Dipylidium caninum were identified. Infections by N. pulicis and Leptomonas sp. occurred mainly in the warm-rainy period. The prevalence and distribution of these endosymbionts in fleas derived from Brazil and South America, and the their variation according to sex and season, are reported for the first time.

Biogeography of Brazilian populations of Panstrongylus megistus (Hemiptera, Reduviidae, Triatominae) based on molecular marker and paleo-vegetational data.

In epidemiological terms, Panstrongylus megistus is one of the most important species of triatomine bug in Brazil. Samples from 11 localities were studied using the random amplification of polymorphic DNA (RAPD) technique, which was able to differentiate the study populations clearly. Biogeographical data indicate that these populations could already have arisen 18,000 years ago (C(14)), it being possible to differentiate insects from the Brazilian states of Santa Catarina (SC) in the south, Ceará (CE) in the northeast and another large intermediate block containing the remaining eight populations from five other states. These results agree with those obtained by phenograms constructed from RAPD data, in which the SC population lies opposite those of CE, consistent with the greatest geographical distance between these localities. The other eight populations (Alagoas (AL), Bahia (BA), Goiás (GO), Minas Gerais (MG) and São Paulo (SP)) are closer genetically and originated in areas whose vegetational characteristics have remained similar to each other during the last 18,000 years, thus allowing greater contact between them. The greatest divergence of this group of insects and those of Ceará appears to have occurred 8000 years ago. This more humid period gave rise to other landscape changes, allowing greater differentiation of the vegetation and consequent expansion of P. megistus populations. Formation of the Serras do Mar and Mantiqueira probably created geographical barriers that favored a certain degree of isolation and greater differentiation of the SC population. Atlantic forest remnants within the caatinga domain (created between 25 and 17,000 years ago), where the CE populations originated probably constitute ecological refugia produced by successive amplification and retraction of the most suitable habitats for this species.

Polygenis (Polygenis) platensis (Jordan & Rothschild) (Siphonaptera: Rhopalopsyllidae, Rhopalopsyllinae), a new record in Brazil

Polygenis (Polygenis) platensis s. l. (Jordan & Rothschild) foi assinalada pela primeira vez no Brasil nos municípios de Capão da Canoa, Tramandaí e Osório, RS. Novos registros de hospedeiros são dados para a espécie, com a inclusão de Ctenomys flamarioni (Travi) e Ctenomys minutus (De Blainville). Variações morfológicas foram também observadas entre os espécimes coletados sobre mesma espécie de hospedeiro e em mesma localidade, evidenciando alguma hibridização entre as duas subspécies e impossibilitando a determinação do status subespecífico. O número de espécies de pulgas conhecidas para o Brasil é aumentado para 60.

Polygenis (Polygenis) platensis s. l. (Jordan & Rothschild) is recorded for the first time from Brazil in the municipalities of Capão da Canoa, Tramandaí and Osório, RS. New hostrecords for this flea include Ctenomys flamarioni (Travi) and Ctenomys minutus (De Blainville). Morphological variations were also observed among the flea specimens collected on the same species of host at the same municipality, evidencing some hybridization between the two subspecies and disabling the determination of the subspecific status. The number of known species of Brazilian Siphonaptera is now 60.

New data on the morphology of Polygenis (Polygenis) rimatus (Jordan) (Siphonaptera: Rhopalopsyllidae)

The study was carried out on the basis of 208 specimens of both sexes from a wide geographical range, from central Brazil to central Argentina. The morphology of diagnostic features of Polygenis (Polygenis) rimatus (Jordan) including the presence or absence of the ductus of the spermatheca with a thick wall, the shape of the posterior margin of seventh sternum in females, and the size of the angle between the distal and the proximal arms of the basal part of the aedeagal tubus, and the number of convolutions made by the coil of the aedeagal tubus interior in males, have been studied. The results obtained are important in clarifying some difficulty in species recognition. Besides, the variability of some characteristics between specimens from disjoint localities in Brazil and Argentina is also noticed.

O estudo baseou-se em 208 espécimes, coletados em uma ampla faixa geográfica situada entre os centros do Brasil e da Argentina. Morfologicamente, as características utilizadas para a identificação de Polygenis (Polygenis) rimatus (Jordan) foram estudadas, tais como a presença ou ausência do duto da espermateca com margens espessadas e a forma da margem posterior do esternito VII, entre as fêmeas, bem como o ângulo entre os braços proximal e distal do tubo interno do aedeagus e o número de suas circunvoluções, nos exemplares machos. Os resultados obtidos são importantes para subsidiarem o reconhecimento da espécie. A variabilidade de algumas características entre exemplares de localidades disjuntas no Brasil e Argentina é também noticiada.

Interpopulation variability among Panstrongylus megistus (Hemiptera: Reduviidae) from Brazil.

Comparisons were made among Panstrongylus megistus (Burmeister) from three areas of Brazil (Bahia, Minas Gerais, and Santa Catarina), where populations differ with regard to their degree of association with human dwellings. The following characters were studied: morphology of the eggs and the male genitalia; morphometry of the head and thorax; isoenzyme profile; enzymatic activity of the alpha-glycerophosphate dehydrogenase (alpha-GPDH); and cytogenetics. In general, differences were observed in the weight, diameter, and length of the egg among Bahia, Minas Gerais, and Santa Catarina populations. Differences were not observed in the architecture of the egg exochorion. The size of the median process of the pygophore of the male genitalia of individuals from Bahia differed from the other two populations. The Minas Gerais population presented the largest number of denticles in the endosome process. The morphometry of the head and thorax differentiated Santa Catarina from the Bahia and Minas Gerais populations. Phosphoglucomutase (EC 5.4.2.2., PGM) was the only enzyme out of 11 that showed polymorphism; the population from Minas Gerais was most polymorphic, whereas the population from Santa Catarina was monomorphic. Study of the alpha-GPDH activity and cytogenetics did not reveal differences among the three populations. Analysis of all the characters studied together with information on these three populations from previous publications allowed a phenogram to be constructed. Two distinct groups were evident, one represented by Santa Catarina and the other by Bahia and Minas Gerais. Considering the greater variability of the Minas Gerais and Bahia populations, we propose the inclusion of these states within the area of origin of P. megistus.

First report of Polygenis (Polygenis) roberti beebei (Fox, 1947) (Siphonaptera: Rhopalopsyllidae) in Argentina, with a new host record and morphological data.

In the present study we report the presence of Polygenis (Polygenis) roberti beebei (Fox, 1947) for the first time in Argentina (23 degrees 56'S, 64 degrees 28'W, Jujuy Province). A new host record (the rodent Oryzomys russatus) and morphological details are also presented. Although the Argentinean specimens fit descriptions and illustrations given previously, the only studied male differs from the specimens collected from other localities in the characteristics of its aedeagus. Collections of further material and morphological studies are needed in order to analyse the taxonomic status of the specimens from Argentina.