Banco de tejidos de la Colección Biológica Grupo de Estudio Relación Parásito Hospedero

Study Extent

The samples are from different regions of Colombia: Antioquia, Bogotá D.C., Caquetá, Casanare, Cundinamarca, Guaviare, Magdalena, Meta, Nariño, Risaralda, Santander and Vichada. which includes many habitats as: flooded savannas, wetlands, tropical savanna, Amazon forest, tropical rainforest, tropical cloud forest, Chocó Biogeographical Region, urban transformed landscapes, Paramo, ecotone Orinoco-Amazon.

Sampling

Sampling methods are according to the vertebrate group, as indicated below. For birds, mist nets were used to capture and individual were mesured, photographed and identified using the field guides Hilty, Brown W, Brown L (1986) and Ayerbe-Quiñonez, F (2019). Blood samples were taken by brachial vein puncture or toenail clip for hummingbirds. The amphibians and reptiles were captured manually after an active search, for may snakes an herpetologic hook was used. As mention before, every individual was mesured, photographed and identified. For taking the blood sample different methodologies were used depending on the biological group. The amphibians were sampled by a Maxillary vein puncture, but the smaller ones were colected and dissected. Many reptiles as the lizards, crocodiles alligators, snakes and iguanas were sampled by a ventral coccygeal vein puncture, and the turtles by a Subcarapacial sinus or coccygeal vein puncture. Lastly bats were captured using mist nets and sampled by Antebrachial or interfemoral vein puncture. After the individual was sampled, the blood were stored in etanol (96%), SET buffer or EDTA, also blood smears were made and fixed in methanol for 5 minutes.

Quality Control

Every field work were accompanied by experts in every taxonomic groups, and the individual photographs were contrasted after the fieldwork to make a correct host idetification. For the haemoparasites, its identification were made by a microscopic diagnosis using the taxonomic key of Valkiunas (2005). Then a molecular diagnosis were made in many cases, by a nested PCR, after a phenol chloroform DNA extraction. Sequencing were made by Macrogen Korea, and the were contrasted with Genbank and/or MalAvi.

Method steps

1. The first stage is the field work in which the hosts (vertebrates) are captured, identified, mesured, photographied and samples as mention before.
2. The next step is the staining of the blood smears with giemsa pH 7.2 for 45 minutes. Now samples are ready for a microscopic analysis using a light microscope. Positive samples are photographid in a Leica DM570 microscope with camera. Parasites are mesured and identified with the taxonomic key of Valkiunas (2005).
3. Then molecular diagnosis is made to positive samples. Phenol chloroform DNA extraction is made to the blood stored. Also, many samples were extracted with the extraction kit Quick-DNA MiniPrep ZYMO. Amplification of different makers is made depending the parasite, for Hepatozoon 18s is used, and the CytB for Apicomplexa parasites. Amplification products are deposited in Genbank and MalAvi.

The resource has a wide diversity of vertebrates samples. The vast majority of records belongs to birds, classified in 44 families from 18 orders. The reptiles and amphibians have fewer records, reptiles samples belongs to the Alligatoridae and Crocodylidae families from the order Crocodylia; also to the Boidae, Colubridae, Dactyloidae, Gekkonidae, Iguanidae, Teiidae, Tropiduridae and Viperidae families from Squamata and the Emydidae, Geoemydidae, Podocnemididae and Testudinidae families from Testudines. The amphibians are represented by the Bufonidae, Craugastoridae, Hylidae, Leptodactylidae, Microhylidae and Phyllomedusidae families. The mammals has the least amount of records, belonging to two species of the Phyllostomidae family, and another two species from Vespertilionidae Many of these vertebrates host haemoparasites that are included in the resource, belonging to the families: Babesiidae, Dactylosomatidae, Haemogregarinidae, Haemoproteidae, Hepatozoidae, Lankesterellidae, Leucocytozoidae, Plasmodiidae, Rickettsiaceae, Trypanosomatidae and the superfamily Filarioidea.

1. Accipitriformes
   common name: hawks, eagles, vultures and kites rank: order
2. Anseriformes
   common name: ducks, geese and swans rank: order
3. Apodiformes
   common name: swifts and hummingbirds rank: order
4. Caprimulgiformes
   common name: Nightjars rank: order
5. Charadriiformes
   rank: order
6. Columbiformes
   common name: doves and pigeons rank: order
7. Coraciiformes
   common name: kingfishers, momots and others rank: order
8. Cuculiformes
   common name: cuckoos rank: order
9. Falconiformes
   common name: falcons and caracaras rank: order
10. Galbuliformes
    rank: order
11. Galliformes
    rank: order
12. Gruiformes
    rank: order
13. Passeriformes
    rank: order
14. Pelecaniformes
    rank: order
15. Piciformes
    rank: order
16. Psittaciformes
    common name: parrots rank: order
17. Strigiformes
    common name: owls rank: order
18. Alligatoridae
    common name: alligators and caimnas rank: family
19. Crocodylidae
    common name: crocodiles rank: family
20. Boidae
    common name: boids rank: family
21. Colubridae
    common name: colubrids rank: family
22. Dactyloidae
    common name: anoles rank: family
23. Gekkonidae
    common name: geckos rank: family
24. Iguanidae
    rank: family
25. Teiidae
    rank: family
26. Tropiduridae
    rank: family
27. Viperidae
    common name: vipers rank: family
28. Emydidae
    rank: family
29. Geoemydidae
    rank: family
30. Podocnemididae
    rank: family
31. Testudinidae
    rank: family
32. Bufonidae
    rank: family
33. Craugastoridae
    rank: family
34. Hylidae
    rank: family
35. Leptodactylidae
    rank: family
36. Microhylidae
    rank: family
37. Phyllomedusidae
    rank: family
38. Phyllostomidae
    rank: family
39. Vespertilionidae
    rank: family
40. Dactylosoma
    rank: genus
41. Haemogregarina
    rank: genus
42. Hepatozoon
    rank: genus
43. Lankesterella
    rank: genus
44. Haemoproteus
    rank: genus
45. Leucocytozoon
    rank: genus
46. Plasmodium
    rank: genus
47. Haemocystidium
    rank: genus
48. Babesia
    rank: genus
49. Rickettsia
    rank: genus
50. Trypanosoma
    rank: genus
51. Filarioidea
    rank: superfamily

This resource includes samples from different regions / ecosystems of Colombia: flooded savannas, wetlands, tropical savanna, Amazon forest, tropical rainforest, tropical cloud forest, The Chocó Biogeographical Region, urban transformed landscapes, Paramo and the ecotone Orinoco-Amazon.