Characterization of the excretory/secretory products of Dermatobia hominis larvae, the human bot fly

Proteolytic activity in excretory/secretory products (ESP) of first- (L1), second- (L2) and third-instar (L3) larvae of Dermatobia hominis was analyzed through gelatin-gel and colorimetric enzyme assays with the chromogenic substrates azocasein and BApNA. The functional characterization of proteases was based on inhibition assays including synthetic inhibitors. ESP were obtained from new-hatched larvae reared in the laboratory and from second- and third-instar larvae removed from naturally infested cattle. Gelatin-gel analysis evidenced few bands of proteolysis, predominantly of high apparent molecular masses, in ESP of L1, whereas in the gel of L2 and U ESP there was a wide range of proteolytic activity most of them not resolved in a single species. Azocasein assays revealed a progressive increase of protease activity from first- to third-instar larvae. Protease inhibitor assays revealed a predominance of metalloproteases in L1 ESP that could be related to a skin penetration process and to a diversion of host immune response. The predominance of serine proteases in L2 and L3 and the great tryptic activity presented by L3 ESP were attributed to an increasing trophic activity by the growing larvae, since the viability of adult flies strictly depends on larval abilities to assimilate nutrients from the host. Taking together, these results suggest that Dematobia larvae secrete/excrete different proteases that may be related to diverse functions during host penetration and infestation, which reinforces the relevance of the study of such proteolytic enzymes. (C) 2009 Elsevier B.V. All rights reserved.

A new workerless inquiline in the Lower Attini (Hymenoptera: Formicidae), with a discussion of social parasitism in fungus-growing ants

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cardiorespiratory responses to hypoxia in the African catfish, Clarias gariepinus (Burchell 1822), an air-breathing fish

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cytogenetics and embryology of Eupatorium laevigatum (Compositae)

Embryological studies indicate Eupatorium laevigatum to have Antennaria type diplospory with precocious embryony. The embryo sac is of the Polygonum type and the polar nuclei fuse before anthesis (maturation of the stamens). Endosperm development is autonomous and the central cell divides only after the initial stages of embryo formation. It is estimated that about 10% of the florets in anthesis contain an undivided egg which can be used for sexual reproduction. The study of microsporogenesis revealed abnormalities in chromosome pairing which result in the formation of univalents, bivalents, trivalents and higher polyvalents, with the consequent production of lagging chromosomes, unbalanced nuclei, micronuclei and sterile pollen. We found that, as represented by the material studied, E. laevigatum is an autohexaploid (2n = 6x = 60) in which each chromosome of a basic set of ten chromosomes is repeated six times and that E. laevigatum is an essentialy obligate apomictic.

Alkyl Hydroxybenzoic Acid Derivatives that Inhibit HIV-1 Protease Dimerization

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Curva de sobrevivência e estimativa de entropia em Lucilia cuprina (Diptera, Calliphoridae)

Lucilia cuprina (Wiedemann, 1830) is a cosmopolite blowfly species of medical and veterinary importance because it produces myiasis, mainly in ovine. In order to evaluate the demographic characteristics of this species, survivorship curves for 327 adult males and 323 adult females, from generation F1 maintained under experimental conditions, were obtained. Entropy was utilized as the estimator of the survival pattern to quantify the mortality distribution of individuals as a function of age. The entropy values 0.216 (males) and 0.303 (females) were obtained. These results denote that, considering the survivorship interval until the death of the last individual for each sex, the males present a tendency of mortality in more advanced age intervals, in comparison with the females.

Prevalence of larvae of the bot fly Cuterebra simulans (Diptera, Oestridae) on Gracilinanus microtarsus (Didelphimorphia, Didelphidae) in southeastern Cerrado from Brazil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Would Chrysomya albiceps (Diptera: Calliphoridae) be a beneficial species?

Chrysomya albiceps (Widemann) é uma mosca que se desenvolve em carcaças e opcionalmente pode causar miíase secundária. Larvas de segundo estádio foram removidas de uma lesão existente em uma ovelha da raça Merino em Botucatu. Entre a lã, ao redor da lesão, foram encontradas larvas de primeiro estádio. Também no interior da lesão foi obtida uma larva de terceiro estádio. As larvas foram mantidas em laboratório e delas obtidos insetos adultos, com 50 casais formados e mantidos em gaiolas por oito gerações. de cada geração, 100 adultos eram sacrificados e examinados morfologicamente, com os seus caracteres confrontados com os de Chrysomya rufifacies. A larva de terceiro estádio deu origem a Cochliomyia hominivorax e as demais a C. albiceps. Foi verificado que C. albiceps, além de ser capaz de danificar tecido integro, é também uma possível predadora de larvas de C. hominivorax. A importância de C. albiceps para os animais domésticos e sua associação com C. hominivorax é aqui discutida.

CELL-MEDIATED AND HUMORAL IMMUNE-RESPONSES IN IMMUNIZED AND OR DERMATOBIA-HOMINIS INFESTED RABBITS

The cell-mediated and humoral immune response of rabbits to antigens from larvae of Dermatobia hominis were analyzed by leucocyte migration inhibition factor assay (MIF), immunodiffusion (ID) and passive hemagglutination (PH) test in rabbits immunized with D. hominis extract, in rabbits immunized and infested with the parasite and rabbits infested with D. hominis. Twenty rabbits were divided into five groups: Group 1, rabbits immunized with a crude antigen extract, evaluated for 40 weeks at 4 week intervals; Group 2, rabbits immunized and infested with newly hatched larvae at 14 weeks post immunization (PI) and evaluated as Group 1; Group 3, rabbits immunized, evaluated for 28 weeks at 2 week intervals; Group 4, rabbits immunized and infested at 4 weeks PI and evaluated as Group 3; Group 5, rabbits infested and evaluated for 24 weeks at 2 week intervals. Different patterns of reactivity were observed in the infested and immunized animals: immunized rabbits developed antibodies and cellular immune responses earlier and at higher levels during immunization than the infested rabbits; the infestation at 14 weeks PI, when the cell-mediated and humoral immune response began to decrease, or at 4 weeks PI when these parameters were at higher levels, elicited an anamnestic response. After the spontaneous elimination of larvae by the host, from the 4th week PI onwards, high titers of antibodies and migration inhibition indices were maintained for a long period. These results suggest that the onset of cellular and humoral immune responses after immunization may be important as a biological control of myiasis and contribute to better understanding of the immune defense mechanism of the host against D. hominis.

A specialized midgut region in stingless bees (Hymenoptera: Apidae: Meliponinae)

Ultrastructure of the anterior end of the midgut of three Neotropical stingless bees, including an obligate necrophagous species Trigona hypogea, suggests that this midgut region has no role in nutrient absorption because its lumen is reduced by a thick cuticle that lines the outer epithelium of the cardiac valve. A possible involvement of this midgut region in the production of the peritrophic membrane compounds is discussed.

Prevalence and molecular characterization of Anaplasmataceae agents in free-ranging Brazilian marsh deer (Blastocerus dichotomus)

Anaplasmataceae organisms comprise a group of obligate intracellular gram-negative, tick-borne bacteria that can infect both animals and humans. In the present work we investigate the presence of Ehrlichia, Anaplasrna, and Neorickettsia species in blood samples from Brazilian marsh deer (Blastocerus dichotomus), using both molecular and serologic techniques. Blood was collected from 143 deer captured along floodplains of the Parana River, near the Porto Primavera hydroelectric power plant. Before and after flooding, marsh deer were captured for a wide range research program under the financial support of São Paulo State Energy Company (CESP), between 1998 and 2001. Samples were divided into four groups according to time and location of capture and named MS01 (n = 99), MS02 (n = 18) (Mato Grosso do Sul, before and after flooding, respectively), PX (n = 9; Peixe River, after flooding), and AGUA (n = 17; Aguapei River, after flooding). The seroprevalences for Ehrlichia chaffeensis and Anaplasma phagocytophilum were 76.76% and 20.2% in MS01, 88.88% and 5.55% in MS02, 88.88% and 22.22% in PX, and 94.12% and 5.88% in AGUA, respectively. Sixty-one animals (42.65% of the total population) were PCR-positive for E. chaffeensis PCR (100.0% identity based on 16S rRNA, dsb, and groESL genes). Seventy deer (48.95% of the total population) were PCR-positive for Anaplasma spp. (99.0% of identity with A. platys, and in the same clade as A. phagocytophilum, A. bovis, and A. platys based on 16S rRNA phylogenetic analysis). Our results demonstrate that Brazilian marsh deer are exposed to E. chaffeensis and Anaplasma spp. and may act as reservoirs for these rickettsial agents, playing a role in disease transmission to humans and other animals. (c) 2012 Elsevier Ltd. All rights reserved.

Rabbit antibody responses to experimental infestation with Dermatobia hominis.

The three larval stages of Dermatobia hominis (Linnaeus) have been evaluated for their immunogenicity by ELISA and immunodiffusion (ID) using sera from experimentally infested rabbits. During a primary infestation, first instar D. hominis were found to cause most reaction and allowed the earliest diagnosis by ELISA. An inhibition of the antibody response against second and third instars was observed. The inhibition disappeared after departure of the larvae from the host. In experimentally immunized hosts the antibody response, following challenge, was highest against second and third instar antigens. Antibody remained elevated during the infestation but fell immediately after the larvae had left the host.

Epidemiology of sheep myiases in São Paulo State, Brazil

A survey of Diptera species causing cutaneous myiases on sheep in Botucatu, São Paulo, Brazil was made to determine seasonal incidence, predilection sites and the factors predisposing to infestation. Sheep were checked daily for myiases for one year. At two week intervals larvae from wounds were collected for identification. Only larvae of Dermatobia hominis and Cochliomyia hominivorax were found. Myiases due to C. hominivorax were observed during the whole year with high incidence from January to April. The feet, vulva, tail and scrotum were most frequently infested. Wounds were the commonest predisposing factor. © 1992 Centre for Tropical Veterinary Medicine, University of Edinburgh.

Oftalmomiíase externa causada por cochliomyia hominivorax

The purpose of this study was to report a case of external ophthalmomyiasis caused by Cochliomyia hominivorax in a 10 years-old child, which presented acute preseptal celulitis and had 17 worms removed from the lacrimal canaliculus and conjunctival cul de sac, bilaterally, with resolution. The authors discuss the diagnosis and treatment of this nosological entity.

Thelytokous parthenogenesis in the fungus-gardening ant Mycocepurus smithii (Hymenoptera: Formicidae)

The general prevalence of sexual reproduction over asexual reproduction among organisms testifies to the evolutionary benefits of recombination, such as accelerated adaptation to changing environments and elimination of deleterious mutations. Documented instances of asexual reproduction in groups otherwise dominated by sexual reproduction challenge evolutionary biologists to understand the special circumstances that might confer an advantage to asexual reproductive strategies. Here we report one such instance of asexual reproduction in the ants. We present evidence for obligate thelytoky in the asexual fungus-gardening ant, Mycocepurus smithii, in which queens produce female offspring from unfertilized eggs, workers are sterile, and males appear to be completely absent. Obligate thelytoky is implicated by reproductive physiology of queens, lack of males, absence of mating behavior, and natural history observations. An obligate thelytoky hypothesis is further supported by the absence of evidence indicating sexual reproduction or genetic recombination across the species' extensive distribution range (Mexico-Argentina). Potential conflicting evidence for sexual reproduction in this species derives from three Mycocepurus males reported in the literature, previously regarded as possible males of M. smithii. However, we show here that these specimens represent males of the congeneric species M. obsoletus, and not males of M. smithii. Mycocepurus smithii is unique among ants and among eusocial Hymenoptera, in that males seem to be completely absent and only queens (and not workers) produce diploid offspring via thelytoky. Because colonies consisting only of females can be propagated consecutively in the laboratory, M. smithii could be an adequate study organism a) to test hypotheses of the population-genetic advantages and disadvantages of asexual reproduction in a social organism and b) inform kin conflict theory. For a Portuguese translation of the abstract, please see Abstract S1. © 2009 Rabeling et al.