Marta Wolff

Isolation and characterization of a novel insect defensin from Rhodnius prolixus, a vector of Chagas disease.

An antimicrobial peptide belonging to the defensin family of small cationic peptides associated with innate immunity in insects was isolated from the hemolymph of Rhodnius prolixus, a vector of Chagas disease. This peptide, designated R. prolixus defensin A, was purified and sequenced. The active peptide contains 43 residues and aligns well with other insect defensins. However the pre-pro region of the sequence has little shared identity with other insect defensins. We have identified 3 isoforms of R. prolixus defensin from cDNA clones obtained from RNA isolated from the whole bodies of immune activated insects. Northern analysis and Real-Time Quantitative PCR indicate that there is a very low baseline transcription of this peptide in naïve insects, and that transcription increases significantly in the fat body of immune activated insects. In addition there is a delayed induction of transcription of this peptide in the intestine 24 h post activation suggesting that the midgut/intestine of this species is active in the immune response against pathogens.

Successional behavior and occurrence matrix of carrion-associated arthropods in the urban area of Medellín, Colombia.

Forensic entomology is a scientific tool applied to the study of insect or arthropod succession at the scene of a crime or that associated with an accident or natural death. Interpreting this succession provides information to determine minimum and maximum limits of the Postmortem Interval (PMI), that is, the time between death and the discovery of the body. This study was carried out during the rainy season, from 27 October to 12 December 2002 in an urban area of the city of Medellín, Colombia. Three domestic pigs were used as animal models. The results showed that both the aforementioned species as well as the physical characteristics of the carcass determined the indicator species of the postmortem interval in urban areas of the city. In total, 11,937 individuals were collected and identified, belonging to 12 orders, 29 families and 42 genera. Diptera were the most abundant order (90%) represented mainly by Calliphoridae (80%). Coleoptera were scarcely present, representing only 2.8% of the total collected. Based on information obtained of eggs and larvae reared to adult of this successional study, an occurrence matrix was elaborated for determining of the PMI. From the same study area and under the same conditions, a list of arthropods associated with carrion is presented in this manuscript.

Identification and characterization of two novel lysozymes from Rhodnius prolixus, a vector of Chagas disease.

Lysozymes have been described in invertebrates as digestive or immune molecules. We report here the characterization of two novel c-type lysozymes, RpLys-A (EU250274) and RpLys-B (EU250275), isolated from the fat body and digestive tract of immune stimulated Rhodnius prolixus, a major vector of Chagas disease. Transcriptional profiles indicate that the temporal and spatial expression patterns of these two peptides are very different. RpLys-A is expressed predominantly in the midgut after ingestion of Trypanosoma cruzi in a bloodmeal, or after injection of bacteria into the hemocoel. RpLys-B is expressed primarily in the fat body after bacterial injection. Phylogenetic alignments indicate that RpLys-A aligns best with molecules from other hemipterans whose major expression is found in the intestinal tract whereas RpLys-B aligns best with mosquito and tick molecules whose expression is found principally in hemocytes and fat body and whose role has been described as immune-related. These data suggest a differential compartmentalized role of two closely related molecules; one for immunity in the hemocoel and the other for digestion in the midgut.

Descripción y clave de los estadios inmaduros de las principales especies de Calliphoridae (Diptera) de importancia forense en Colombia

Larvae of 13 blowfly species from Colombia are described and an illustrated key for all them is presented. All larval instars of Calliphora nigribasis Macquart, Cochliomyia macellaria (Fabricius), Chrysomya albiceps (Wiedemann), Chrysomya megacephala (Fabricius), Hemilucilia segmentaria (Fabricius), Hemilucilia semidiaphana (Rondani), Lucilia eximia (Weidemann) are described, but the second and third instars of Compsomyiops verena (Walter), and only the third instar of Calliphora vicina Robineau-Desvoidy, Cochliomyia hominivorax (Coquerel), Lucilia peruviana Robineau-Desvoidy, Lucilia sericata (Meigen) and Sarconesiopsis magellanica (Le Guillou).

Initial study of arthropods succession and pig carrion decomposition in two freshwater ecosystems in the Colombian Andes

Entomological succession and trophic roles of arthropods associated with different stages of carcass decomposition were studied to estimate the post-mortem submersion interval in two freshwater ecosystems in the Colombian Andes, at an altitude of 2614 m. Pig carcasses were employed as models placed 68 m apart, one in a stream (lotic) and another in an artificial lake (lentic). Decomposition time to skeletal remains was 74 days in the lake and 80 days in the stream. Six phases of decomposition were established: submerged fresh, early floating, floating decay, bloated deterioration, floating remains and sunken remains. A total of 18,832 organisms associated with the carcasses were collected: 11,487 in the lake (four orders, 19 families and 33 species) and 7345 in the stream (eight orders, 15 families and 25 species). Organisms were classified in the following ecological categories: shredders, collectors, predators, necrophagous, sarcosaprophagous and opportunists. Physical and chemical properties of the habitats, such as water temperature, CO(2) and conductivity, varied according to rainfall. In the lake, shredders (Coleoptera: Tropisternus sp. and Berosus sp.) and collectors (Diptera: Chironomus sp.) were found to be associated with submerged phases. Predators (Odonata) were only present during the first phases. Coleoptera (Dytiscidae) were found during floating decay and bloated deterioration stages. In the stream, shredders (Hyalella sp.) and collectors (Simulium sp.) were found during all stages, whereas the predator Oxelytrum discicolle was found exclusively during the floating stages, during which body temperature increased in a fashion similar to active decay in terrestrial environments.

Domiciliation Trend of Panstrongylus rufotuberculatus in Colombia

The present paper presents evidence of the domiciliation of Panstrongylus rufotuberculatus in La Gardenia, Colombia through the collection of 2 unhatched eggs, 81 nymphs and 10 adults (4 males and 6 females), from 2 rural houses. The transmission risk indicators of Trypanosoma cruzi by P. rufotuberculatus in La Gardenia, were: domiciliary infestation 7.5%, density 2.35, colonization 66.6%, overcrowding 31.33, natural infection 4.6%, and relative infection 2.5%. These results and findings in Peru and Argentina, show that P. rufotuberculatus has a potential success in domiciliation and could some day become an alternate vector of American trypanosomiasis.

Rearing five species of Diptera (Calliphoridae) of forensic importance in Colombia in semicontrolled field conditions

AbstRACt The family Calliphoridae is widely known to lead the colonization of corpses and their development rates are frequently used to estimate the postmortem interval. This study presents the larval growth of five forensically important species of Calliphoridae in Colombia. Rearing took place in semicontrolled field conditions where the egg masses were collected. We show curves of larval growth, larval length and time intervals to reach all immature stages for Lucilia eximia and Cochliomyia macellaria at two sites with different climatic conditions and for Chrysomya albiceps, Chrysomya megacephala and Calliphora nigribasis at one site. Overall, high temperatures speeded up the development of the species reared at two different sites, whereas low temperatures for C. nigribasis, lengthened the total development time. Differences between this study and others can be explained by the experimental conditions in the field without the possibility of strict laboratory rearing controls.

Description of Pintomyia limafalcaoae and Pintomyia antioquiensis, Two New Species of Phlebotomine Sand Fly (Diptera, Psychodidae) from the Colombian Andes

Two new species of phlebotomine sand fly from Colombian Andes are described, belonging to the subgenus Pifanomyia of the genus Pintomyia. P. (P.) limafalcaoae sp. nov. for which both sexes are described, is assigned to the series pia while P. (P.) antioquiensis sp. nov., known only from the male, is included in the series verrucarum. The subgenus Pifanomyia is characterized and identification keys presented for the two new species.

Synanthropy and ecological aspects of Muscidae (Diptera) in a tropical dry forest ecosystem in Colombia

The synanthropic index and other ecological aspects of the Muscidae family were evaluated through simultaneous monthly sampling in three different environments (urban, rural and forest) using van Someren-Rydon traps baited with human faeces, chicken viscera and decomposing fish and onion. Four traps were set up in each environment (one per bait item) for 48 hours per month, with samples taken every 12 hours. A total of 5726 specimens were collected, belonging to 19 species and 13 genera. Brontaea normata (+99,9), Brontaea quadristigma (+96,9), Synthesiomyia nudiseta (+96,5), Ophyra aenescens (+96,2), Musca domestica (+95,7) and Atherigona orientalis (+93,8) had the highest synanthropic indices, showing a marked preference for human environments. The most abundant species were B. normata (24,31%), Biopyrellia bipuncta (20,60%) and Pseudoptilolepis nigripoda (15,82%), the latter two showed a preference for uninhabited areas. A total of 11 new records for Colombia were found: Ophyra aenescens, Cyrtoneuropsis pararescita, Morellia basalis, Neomuscina dorsipuncta, Biopyrellia bipuncta, Pseudoptilolepis nigripoda, Neomuscina instabilis, Neomuscina currani, Polietina orbitalis, Neomuscina pictipennis and Cyrtoneuropsis maculipennis. Except for the first four species, the remainder presented negative synantrophy indexes (from minor to major), which would allow to use them as ecological indicators of the disturbance degree of dry forests in Colombia.

Insects associated with the composting process of solid urban waste separated at the source

Sarcosaprophagous macroinvertebrates (earthworms, termites and a number of Diptera larvae) enhance changes in the physical and chemical properties of organic matter during degradation and stabilization processes in composting, causing a decrease in the molecular weights of compounds. This activity makes these organisms excellent recyclers of organic matter. This article evaluates the succession of insects associated with the decomposition of solid urban waste separated at the source. The study was carried out in the city of Medellin, Colombia. A total of 11,732 individuals were determined, belonging to the classes Insecta and Arachnida. Species of three orders of Insecta were identified, Diptera, Coleoptera and Hymenoptera. Diptera corresponding to 98.5% of the total, was the most abundant and diverse group, with 16 families (Calliphoridae, Drosophilidae, Psychodidae, Fanniidae, Muscidae, Milichiidae, Ulidiidae, Scatopsidae, Sepsidae, Sphaeroceridae, Heleomyzidae, Stratiomyidae, Syrphidae, Phoridae, Tephritidae and Curtonotidae) followed by Coleoptera with five families (Carabidae, Staphylinidae, Ptiliidae, Hydrophilidae and Phalacaridae). Three stages were observed during the composting process, allowing species associated with each stage to be identified. Other species were also present throughout the whole process. In terms of number of species, Diptera was the most important group observed, particularly Ornidia obesa, considered a highly invasive species, and Hermetia illuscens, both reported as beneficial for decomposition of organic matter.