Forensic Science International 212 (2011) 164–172

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Forensic Science International

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Initial study of arthropods succession and pig carrion decomposition in two

freshwater ecosystems in the Colombian Andes
Maria Barrios 1, Marta Wolff *
Grupo de Entomologı´a Universidad de Antioquia, Medellı´n, Colombia

A R T I C L E I N F O A B S T R A C T

Article history: Entomological succession and trophic roles of arthropods associated with different stages of carcass
Received 11 November 2009 decomposition were studied to estimate the post-mortem submersion interval in two freshwater
Received in revised form 1 June 2011 ecosystems in the Colombian Andes, at an altitude of 2614 m. Pig carcasses were employed as models
Accepted 2 June 2011
placed 68 m apart, one in a stream (lotic) and another in an artificial lake (lentic). Decomposition time to
Available online 7 July 2011
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
Keywords:
sunken remains. A total of 18,832 organisms associated with the carcasses were collected: 11,487 in the
Forensic entomology
Post-mortem submersion interval
lake (four orders, 19 families and 33 species) and 7345 in the stream (eight orders, 15 families and 25
Aquatic insect succession 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, CO2 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.
ß 2011 Elsevier Ireland Ltd. All rights reserved.

Forensic entomology is a frequently employed tool to deter-
mine the time elapsed between death and discovery of a body
(known as post-mortem interval or PMI), where the sequence of
appearance of different groups of insects is associated with
particular climatic conditions [1,2]. The history of forensic
entomology, from the earliest records of its application in 13th
Century China to its modern use in North America, Europe and
Asia, has been well recorded [3]. In South America, forensic
entomology has evolved significantly during the 21st century [4–
7]. As a result, there is wider knowledge of insect species
associated with decaying cadavers, as well as the duration of
the different stages of decomposition, at different times of the year
and in different bioclimatic zones [8–11].
In general, studies of insects associated with cadavers have
been conducted in terrestrial environments, with just a few studies
carried out in aquatic ecosystems in temperate zones [12–24].
Colonisation of a cadaver by aquatic invertebrates depends on
abiotic factors, such as the physical and chemical properties of the
water, depth and strength of flow, and biotic factors, such as the
fauna and flora where the cadaver may be used as a food source or
for protection [14,18].
In Colombia, forensic entomology is a recent discipline [4,9–
11,25–27], where factors relating to the post-mortem submersion
interval (PMSI) have not yet been fully explored. This study
represents the first attempt to determine entomological succes-
sion and the trophic roles of arthropods associated with different
stages of carcass decomposition in two freshwater ecosystems
(lake and stream) using pig (Sus scrofa) carcasses in a rural zone in
the high Andes. Although data were not replicated, the study
represents a unique starting point to stimulate further studies of
decomposition in aquatic environments to estimate PMSI.
1. Methods

* Corresponding author. Tel.: +57 4 219 5619; fax: +57 4 219 5622.
E-mail addresses: mariabape@gmail.com, mwolff@matematicas.udea.edu.co
(M. Wolff).
1
Permanent address: Calle 67 No. 53-108, Medellı́n, Colombia.
The study was conducted from January to April 2007, in a rural area of the
municipality of Facatativa in the Colombian Andes (48500 3000 N; 748210 700 W) at an
altitude of 2614 m. The study area has an average annual temperature of 13 8C and a
bimodal rainfall pattern. [28] One week prior to placing the corpses at the study
sites, physical and chemical variables of the water were recorded, and sampling of
entomofauna was begun. Two 20-kg pigs were used to model human cadavers,

0379-0738/$ – see front matter ß 2011 Elsevier Ireland Ltd. All rights reserved.
doi:10.1016/j.forsciint.2011.06.008
 M. Barrios, M. Wolff / Forensic Science International 212 (2011) 164–172
killed at the study site by a bullet wound to the thoracic zone from a 9-mm firearm
(by members of the Forensic Science Faculty of the National Police Force). One
carcass was placed in the shade at the side of a stream (1.47 m wide during the dry
season and at an average depth of 70 cm) and the other in a lake (with an area of
37 m [2] and an average depth of 4 m). Each carcass was protected by a metal cage
measuring 100 cm  70 cm  50 cm with a mesh size of 1.5 cm [11], allowing
submersion and floatation of the corpses. Three sampling points were established in
the stream: (1) 3 m upstream from the carcass; (2) at the site of the submerged
carcass; and (3) 3 m downstream from the carcass (points 1 and 3 were used as
controls). Similarly, three sampling points were established in the lake: (a) 3 m to
the left of the carcass; (b) at the site of the carcass; and (c) 3 m to the right of the
carcass (points (a) and (c) were used as controls also). Samples were taken twice a
day, between 8.00–11.00 and 15.00–18.00, from the time of death until just the
skeleton remained. With each sample, the state of the carcass was documented,
recording corporal rectal temperature as well as the physical and chemical
properties of the water, such as dissolved oxygen (digital oximeter Okaton), CO2
(phenolphthalein method), pH (pH measurer Okaton), conductivity (digital
conductimeter Okaton) and water and air temperature (digital thermometer
Okaton). Data on rainfall (mm) were obtained from the meteorological station at
Altamira, at an altitude of 2636 m and 15 km from the study site. For a period of
45 min, arthropods were collected manually with forceps from natural orifices, the
gunshot wound and from the whole body surfaces of both carcasses, without taking
them out of the water. Organisms found around the carcass were collected with a D-
net and nylon nets in the lake and with a screen barrier net in the stream.
Subsequently, all samples were fixed in 80% ethanol, and the solution was changed
after 24 h. Adult specimens were preserved dry and deposited in entomological
boxes. The immature stages were conserved in 80% ethanol in glass vials. Fly larvae
were placed in hot water (90 8C) for 5 min and then preserved in 80% ethanol.
Ecological categories were established using the classification of functional groups
proposed by Merritt and Cummins [29] for aquatic species: shredders (detritivores,
chewers of particulate organic matter (CPOM)), collectors (detritivores, filterers or
suspension feeders of fine particulate organic matter (FPOM)), scrapers (herbivores,
grazing scrapers of mineral and organic surfaces) and predators (engulfers,
carnivores, attack prey and ingest whole animals or parts). Terrestrial species were
classified as necrophagous, sarcosaprophagous or incidental organisms, as
proposed by Smith. [1] Stages of decomposition were determined according to
those set out by Payne and King [13] and Merritt and Wallace [30].
1.1. Statistical analysis
A multivariate analysis (multivariate analysis of variance (MANOVA), P < 0.05;
Statistical Package for Social Sciences (SPSS) 11) was used to evaluate the physical
and chemical variables (water temperature, dissolved oxygen, CO2, pH and
conductivity) at control points 1/a and 3/c, with the stream and lake study areas
representing the sampling universe, given that each decomposition process is
specific to a site.
2. Results
A total of 18,832 organisms associated with submerged
carcasses were collected: 11,487 from the lake (four orders, 19
families and 33 species) (Table 1) and 7345 from the stream (eight
orders, 15 families and 25 species) (Table 2). In the lake were
collected 7123 and 5743 organisms from points ‘a’ (3 m to the left
of the carcass) and ‘c’ (3 m to the right of the carcass) (Table 1) and
in the stream 4870 and 5381 organisms from the points ‘10 (3 m
upstream from the carcass) and ‘30 (3 m downstream from the
carcass) (Table 2). Total PMSI, from sacrifice until skeletal remains,
was 74 days in the lake and 80 in the stream. Six phases of
decomposition were observed: submerged fresh, early floating,
floating decay, bloated deterioration, floating remains and sunken
remains [13,30].
2.1. Submerged Fresh (days 1–8 in the stream and 1–5 in the lake)
The carcasses remained totally submerged in both habitats,
showing a drop in body temperature (to reach water temperatures
within 8 h of death in the lake and within 3 h in the stream) and
stiffening, which began in the mouth and upper extremities and
was present throughout this stage. The organisms found during
this stage were exclusively aquatic; the most abundant were
collectors: 48% in the lake, represented principally by Chironomus
sp. and 54% in the stream, represented by Simulium sp. Shredders,
such as Tropisternus sp. and Hyallela sp. (30% in the lake and 46% in
165
the stream), were also observed. In addition, predators were
observed in the lake (22%) (dragonflies, damselflies and hemi-
pterans) (Table 3).
2.2. Early Floating (days 9–24 in the stream and 6–17 in the lake)
This stage was marked by an evident putrid smell and lividity in
the abdominal region, which later became discoloured with the
onset of putrefaction. During this stage, predators (43%) and
collectors (42%) displaced the shredders (7%) in the lake, whereas
in the stream, collectors (47%) and shredders (40%) remained the
most abundant species. Predators (2%) and necrophages (11%)
appeared for the first time (Table 3). In the lake, this stage was
characterised by the presence of Rhantus sp., Copelatus sp.,
Oxelytrum discicolle (Coleoptera and Silphidae) and some dragon-
flies. In the stream, O. discicolle and Chironomidae larvae were
found for the first time during this stage and were present until the
penultimate stage (floating remains). Adults and eggs of Calliphora
nigribasis (Calliphoridae) were observed on the carcasses in both
habitats during this stage, leading to colonisation of the carcasses
with terrestrial carrion-feeding dipterans. Both carcasses showed a
bloating of the abdomen and increased corporal volume, causing
flotation and subsequent exposition to the terrestrial environment.
2.3. Floating decay (days 25–43 in the stream and days 18–40 in the
lake)
In both carcasses, yellowish fluids and gases were observed
coming out of the anus, with expulsion of the intestine, detachment
of the hoofs and black pigmentation on the hind limbs. Community
composition changed at this stage (Table 3); necrophages became
the most abundant in both habitats (33% in the lake and 37% in the
stream), displacing the other groups. In the lake, they were
accompanied by a greater proportion of predators and collectors
(32% and 20%, respectively), shredders (7%) and opportunist species
(1%). Sarcosaprophages appeared for the first time in both habitats
but at low percentages (7% and 10%). Predators remained at low
percentages in the stream (3%), whereas collectors and shredders
were evident at similar percentages (26% and 24%, respectively).
2.4. Bloated deterioration (days 44–51 in the stream and 41–48 in the
lake)
In both carcasses, openings were observed in the soft tissues
where gases had escaped, leading to the carcass collapsing in on
itself, becoming flaccid and floating on the surface with
disarticulated posterior limbs. In the lake, necrophages were no
longer observed, having been replaced by collectors which became
the most abundant group again (41%), followed by predators (28%)
and shredders (26%). However, in the stream, shredders (44%)
displaced the necrophages, which remained at a low percentage
(13%), but collectors, sarcosaprophages and predators maintained
their previous relative proportions (31%, 2% and 8%, respectively)
(Table 3).
2.5. Floating remains (days 52–61 in the stream and 49–60 in the
lake)
This stage was characterised by just the exposed (floating)
abdominal parts of the carcasses remaining, which subsequently
dried and hardened. Gyrinus sp. (Gyrinidae: Coleoptera) and
Stegobium sp. (Anobiidae: Coleoptera) were observed for the first
time on these remains. In both habitats, collectors were again the
most abundant (49% in the lake and 47% in the stream), followed by
shredders (28% and 30%) and predators (23% and 23%). No other
groups were present (Table 3).
 166
Table 1
Insect succession on a 20 kg pig carcass and insects collected at the control sampling points in a lake in the Colombian Andes.

Ecological category Order Family Species Insect succession on a 20 kg pig carcass Insects Insects
collected 3 m collected
to the left 3 m to the
of the carcass right of the
carcass

SF days 1–5 EF days 6–17 FD days BD days FR days SR days J A J A

18–40 41–49 50–60 61–74

J A J A J A J A J A J A

Shredders Coleoptera Hydrophilidae Tropisternus sp.                

M. Barrios, M. Wolff / Forensic Science International 212 (2011) 164–172

Berosus sp.        

Collectors Diptera Chironomidae Chironomus sp. 1       
Chironomus sp. 2    
Chironomus sp. 3    

Predators Odonata Libellulidae Erythrodiplax sp. 1     

Erythrodiplax sp. 2    
Erythrodiplax sp. 3  
Aeshnidae Rionaeschna sp.   
Coenagrionidae Acanthagrion sp.    
Hemiptera Corixidae Centrocorisa sp.        
Notonectidae Notonecta sp.        
Buenoa sp.    
Coleoptera Dytiscidae Rhantus sp.  
Copelatus sp. 1  
Copelatus sp. 2 
Staphylinidae Ambodina sp   
Gyrinidae Gyrinus sp. 
Silphidae Oxelytrum discicolle    

Necrophages Diptera Calliphoridae Compsomyiops verena 

Calliphora nigribasis   
Sarconesiopsis magellanica 
Chrysomya albiceps 
Sarcophagidae Helicobia sp. 
Sarcophagidae 

Sarcosaprophages Diptera Muscidae Hydrotaea villosa 

Musca domestica 
Trichomorellia nigribita  
Fanniidae Fannia sp.  

Incidentals Coleoptera Scarabaeidae Canthidium sp. 

Anobiidae Stegobium sp. 
Melolonthidae Not identified 
Curculionidae Not identified  

SF: submerged fresh; EF: early floating; FD: floating decay; BD: bloated deterioration; FR: floating remains; SR: sunken remains; J: juvenile stadium (eggs or larvae); A: adult. : if species was recorded on up to 33% of days; : for
species recorded on 34–66% of days; : for species recorded on 67–100% of days.
Table 2
Insect succession on a 20 kg pig carcass and insects collected at the control sampling points in a stream in the Colombian Andes.

Ecological category Order Family Species Insect succession on a 20 kg pig carcass Insects Insects
collected 3 m collected 3 m
upstream downstream
from the from the
carcass carcass

M. Barrios, M. Wolff / Forensic Science International 212 (2011) 164–172

SF days 1–8 EF days FD days BD days FR days SR days J A J A
9–24 25–43 44–51 52–61 62–80

J A J A J A J A J A J A

Shredders Amphipoda Hyalellidae Hyalella sp.        
Ephemeroptera Leptophlebiidae Terpides sp.  

Collectors Coleoptera Elmidae Heterelmis sp.             

Trichoptera Polycentropodidae Polycentropus sp.    
Diptera Simuliidae Simulium sp.        
Chironomidae Not identified     

Predators Odonata Megapodagrionidae Teinopodagrion sp.  

Coleoptera Staphylinidae Phylonthus sp.   
Silphidae Oxelytrum discicolle    
Decapoda Pseudothelphusidae Neostrengeria macropora  
Predators/parasite Hymenoptera Ichneumonidae Not identified  
Necrophages Diptera Calliphoridae Compsomyiops verena  
Calliphora nigribasis   
Calliphora vicina  
Sarconesiopsis magellanica    
Lucilia peruviana 
Chrysomya albiceps 
Chrysomya megacephala 
Sarcophagidae Helicobia sp. 

Sarcosaprophages Diptera Muscidae Ophyra albuquerquei 

Ophyra aenescens 
Hydrotaea villosa 
Muscado mestica 
Trichomorellia nigritibia 
Fanniidae Fannia sp. 

SF: submerged fresh; EF: early floating; FD: floating decay; BD: bloated deterioration; FR: floating remains; SR: sunken remains; J: juvenile stadium (eggs or larvae); A: adult. : if species was recorded on up to 33% of days; : for
species recorded on 34–66% of days; : for species recorded on 67–100% of days.

167
 168
Table 3
Proportion of each ecological group collected at each stage of decomposition.

Decomposition stage Lake Stream

Ecological Percentage % Percentage % insects Percentage % insects Ecological Percentage % Percentage % insects Percentage % insects
category insect succession on collected 3 m to the collected 3 m to the category insect succession on collected 3 m upstream collected 3 m downstream

M. Barrios, M. Wolff / Forensic Science International 212 (2011) 164–172

a 20 kg pig carcass left of the carcass right of the carcass a 20 kg pig carcass from the carcass from the carcass

Submerged fresh Collector 48 (n = 271) 33 (n = 342) 22 (n = 234) Collector 54 (n = 428) 77 (n = 632) 26 (n = 281)
Shredder 30 (n = 170) 33 (n = 637) 70 (n = 738) Shredder 46 (n = 365) 23 (n = 194) 74 (n = 789)
Predator 22 (n = 124) 4 (n = 31) 8 (n = 86)

Early floating Predator 43 (n = 792) 13 (n = 144) 3 (n = 29) Collector 47 (n = 728) 32 (n = 249) 30 (n = 189)
Collector 42 (n n = 774) 14 (n = 152) 40 (n = 372) Shredder 40 (n = 620) 68 (n = 534) 70 (n = 443)
Necrophages 8 (n = 147) 0 (n = 0) 0 (n = 0) Necrophages 11 (n = 170) 0 (n = 0) 0 (n = 0)
Shredder 7 (n = 129) 73 (n = 834) 57 (n = 534) Predator 2 (n = 31) 0 (n = 0) 0 (n n = 0)

Floating decay Necrophages 33 (n = 1804) 0 (n = 0) 0 (n n = 0) Necrophages 37 (n = 1323) 0 (n = 0) 0 (n = 0)

Predator 32 (n = 1750) 15 (n = 263) 10 (n = 104) Collector 26 (n = 929) 45 (n = 353) 53 (n = 390)
Collector 20 (n = 1094) 55 (n = 978) 38 (n = 398) Shredder 24 (n = 858) 55 (n = 437) 47 (n = 342)
Sarcosaprophages 7 (n = 383) 0 (n = 0) 0 (n = 0) Sarcosaprophages 10 (n = 358) 0 (n = 0) 0 (n = 0)
Shredder 7 (n = 383) 30 (n = 534) 52 (n = 532) Predator 3 (n = 107) 0 (n = 0) 0 (n = 0)
Opportunist 1 (n = 55) 0 (n = 0) 0 (n = 0)

Bloated deterioration Collector 41 (n = 745) 78 (n = 866) 39 (n = 375) Shredder 44 (n = 332) 60 (n = 452) 67 (n = 634)
Predator 28 (n = 509) 8 (n = 87) 16 (n = 153) Collector 32 (n = 242) 40 (n = 296) 33 (n = 313)
Shredder 26 (n = 472) 30 (n = 534) 45 (n = 432) Necrophages 13 (n = 98) 0 (n = 0) 0 (n = 0)
Opportunist 3 (n = 55) 0 (n = 0) 0 (n = 0) Predator 9 (n = 68) 0 (n = 0) 0 (n = 0)
Sarcosaprophages 2 (n = 36) 0 (n = )0 0 (n = 0) Sarcosaprophages 2 (n = 15) 0 (n = 0) 0 (n = 0)

Floating remains Collector 49 (n = 683) 18 (n = 167) 52 (n = 430) Collector 47 (n = 112) 57 (n = 450) 60 (n = 503)
Shredder 28 (n = 390) 68 (n = 631) 42 (n = 352) Shredder 30 (n = 71) 43 (n = 342) 40 (n = 342)
Predator 23 (n = 321) 14 (n = 125) 6 (n = )52 Predator 23 (n = 55) 0 (n = 0) 0 (n = 0)

Sunken remains Collector 47 (n = 188) 25 (n = 232) 30 (n = 281) Shredder 63 (n = 274) 51 (n = 479) 72 (n = 832)
Shredder 30 (n = 120) 63 (n = 598) 59 (n = 547) Collector 37 (n = 161) 49 (n = 452) 28 (n = 323)
Predator 23 (n = 92) 12 (n = 112) 11 (n = 94)

n = number of specimens collected.

 M. Barrios, M. Wolff / Forensic Science International 212 (2011) 164–172
2.6. Sunken remains (days 62–80 in the stream and 61–74 in the lake)
The sunken remains in the lake had a viscous appearance until
the water levels increased, covering them completely and
impeding visibility. Remains stayed submerged until the end of
the experiment. In the stream, the carcass was reduced to bones,
which were then swept away by the strong currents. Therefore,
samples were not taken from the skeleton at the end of the
decomposition process. At the end of the decomposition process,
the associated insect community in both habitats was made up of
the same ecological groups as at the beginning of the process: in
the lake, collectors were the most abundant (47%), followed by
shredders (30%) and predators (23%), whereas in the stream,
shredders became the most abundant again (63%), displacing
collectors (37%) (Table 3).
2.7. Succession tables
In the lake, Tropisternus sp., Berosus sp., Chironomus sp.,
Centrocorisa sp. and Notonecta sp. were observed throughout the
decomposition process. Erythrodiplax sp1 and Acanthagrion sp.
were present from the submerged fresh stage until floating decay.
Ambodina sp. and O. discicolle were found during the stages of
floating decay, bloated deterioration and floating remains, whereas
Rhantus sp. and Copelatus sp. were only found during floating decay
and bloated deterioration stages (Table 1). In the stream, Hyallela
sp., Heterelmis sp. and Simulium sp. were found during the whole
process, O. discicolle was present from early floating until floating
remains stages and Polycentropus sp. just from floating decay until
sunken remains. Teinopodagrion sp. and C. nigribasis were
associated with the floating decay stage (Table 2). In the sampling
points in the lake were observed Tropisternus sp., Berosus sp.,
Chironomus sp., Centrocorisa sp., Notonecta sp., Buenoa sp.,
Erytrodiplax sp. and Acanthagrion sp. (Table 1). In the sampling
points in the stream were observed Hyallela sp., Heterelmis sp.,
Simulium sp., Terpides sp., Polycentropus sp. and Chironumus sp.
(Table 2).
Multivariate analysis of variance (MANOVA) showed significant
differences between both habitats in all physical and chemical
variables (P < 0.005). However, differences were only found
between water temperature (P < 0.05), CO2 (P < 0.05) and
conductivity (P < 0.05) between the three sampling sites in each
habitat. Only the carcass temperature, as an indirect measure of
Fig. 1. Rainfall, body and water temperature. Lake. FS: submerged fresh; F tem: early floating; F tar: floating decay; DF: bloated deterioration; RF: floating remains; RS: sunken
remains.
169
the decomposition process, and rainfall, as a general environmen-
tal factor, were compared between both habitats. Body tempera-
ture in both carcasses dropped in the first 72 h to match ambient
water temperature. During the floating decay stage, body
temperature increased to 1 or 2 8C above that of the water
temperature in the lake and 2–3 8C in the stream. At the end of the
floating decay stage in the lake, rainfall increased and body
temperature decreased (Fig. 1). In the stream, body temperature
dropped to that of the water at the beginning of the bloated
deterioration stage, coinciding with an increase in rainfall (Fig. 2).
In the lake, CO2 and conductivity showed similar patterns,
increasing during the floating decay stage, decreasing during
bloated deterioration and floating remains but increasing again
during the submerged remains stage, when the highest values of
conductivity and rainfall were recorded (Fig. 3). In the stream,
these variables maintained relatively constant ranges throughout
the decomposition process and only increased at the end of the
floating remains stage, with increased rainfall (Fig. 4).
3. Discussion
Six stages of decomposition were observed [13,20,30], con-
firming that, in freshwater ecosystems, the decomposition process
is different from that in terrestrial environments where five stages
have been reported [1]. Stages lasted for similar time periods in
both habitats (overall < 10% difference), although with a slightly
longer (but overlapping) decomposition period at each stage in the
stream. Hobischak [20] also recorded longer decomposition times
in stream systems than in lake systems, but decomposition times
were longer in both systems in his study than in ours, probably due
to the effect of the lower water temperatures in Canada favouring
the conservation of the carcass and because the differences
between lake and stream were greater in his study. Wipfli et al.
[31] suggest that changes in temperature in temperate regions
drastically increase the strength of currents, principally in areas of
rapids, accelerating the decomposition process to rates faster than
those in terrestrial environments, given that the force of the water
becomes a mechanical means of detaching soft tissues and bones.
Thus, each decomposition process, both in aquatic and terrestrial
environments, is unique and influenced by the environmental
conditions of each region, specific fauna and habitat and properties
of the carcass itself [22]. The duration of each stage was
determined according to physical changes in the carcass and
170 M. Barrios, M. Wolff / Forensic Science International 212 (2011) 164–172

T water

T body

Rainfall

25 30
FS F Tem F Tar DF RF RS

body and water temperature (°C)

23
25
21

Rainfall (mm)
19 20
17
15
15
13 10
11
5
9
7 0
1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69 73 77

Days

Fig. 2. Rainfall, body and water temperature. Stream. FS: submerged fresh; F tem: early floating, F tar: floating decay; DF: bloated deterioration; RF: floating remains; RS:
sunken remains.

T body
CO2
Conductivity
30 110
Body temperature (°C) and CO2 (mg/l)

FS
F Tem F Tar DF RF RS
25 100

Conductivity (μS/cm)
20 90

15 80

10 70

5 60

0 50
1 4 7 10 13 16 19 22 25 28 31 34 37 40 43 46 49 52 55 58 61 64 67 70 73
Days

Fig. 3. Body temperature, CO2 and conductivity. Lake. FS: submerged fresh; F tem: early floating; F tar: floating decay; DF: bloated deterioration; RF: floating remains; RS:
sunken remains.

T body

CO2

Conductivity
Body temperature (°C) and CO2 (mg/l)

30 F Tar 55
FS F Tem DF RF RS
25 53
Conductividad μS/cm

20 51

15 49

10 47

5 45

0 43
1 5 9 13 17 21 25 29 33 37 41 45 49 53 57 61 65 69 73 77

Days

Fig. 4. Body temperature, CO2 and conductivity. Stream. FS: submerged fresh; F tem: early floating; F tar: floating decay; DF: bloated deterioration; RF: floating remains; RS:
sunken remains.
 M. Barrios, M. Wolff / Forensic Science International 212 (2011) 164–172
changes in its temperature. In this study, the submerged fresh
stage lasted for 7 days in the stream and 4 in the lake, similar to
that recorded by Schultenover and Wallace [32] and different from
that estimated for higher latitudes, from 11 to 13 days in Canada
[20]. The early floating stage lasted 15 days in the stream and 11
days in the lake, different from that reported in Canada in the fall,
at 23 days (stream) and 37 days (lake), suggesting again that the
seasons in temperate regions play a determining role in the
duration of each stage. However, in the tropics, the bimodal rainfall
pattern establishes dry and rainy seasons at different times of the
year. The floating decay stage is possibly the most variable stage in
terms of duration, compared with that reported in others’ studies
[20,22,23,33]. In the present study, it was longer than the other
stages of decomposition in both habitats (18 days in the stream
and 22 in the lake), despite being influenced by the feeding activity
of necrophages, accelerating the rate of decomposition. The
bloated deterioration stage was short and lasted the same amount
of time in both habitats (7 days), differing from that shown by
Hobischack [20], who recorded a time of 28 days. With regard to
the floating remains and sunken remains stages, Payne and King
[13] demonstrated that bacteria and fungi also cause decomposi-
tion; therefore, the duration of the stage should depend on the
environmental conditions where the body is found. However, in
this study, the hypothesis could not be corroborated because the
carcasses were not totally reduced to bones in their original
location due an increase in rainfall. Subsequent higher water levels
in both habitats and stronger currents in the stream increased the
rate of detachment of the bones and caused them to be swept
away. After analysing the SPMI from several different studies,
Knight [34] concluded that the decomposition of a submerged
body in aquatic environments takes approximately twice as long as
in terrestrial environments. This is due to the lower temperatures
of the water and the inhibition of necrophagous insect activity,
given that aquatic arthropods do not show feeding behaviour
within this ecological category. However, decomposition of dead
organic matter is a biological process which involves detritovores,
and, in the case of submerged bodies in freshwater environments,
the effect of macroinvertebrates on the decay process should not
be ruled out [35].
3.1. Ecological succession
In the initial stages of decomposition, carcasses enter a
freshwater ecosystem as particulate organic matter, eventually
becoming inorganic matter through microbial activity [36]. This
transformation releases different nutritional resources over time,
allowing certain functional groups involved in the process of
organic decay in aquatic ecosystems to participate in the
decomposition process. Therefore, freshwater arthropods have
the potential to be used as a tool to calculate PMSI. Hobischak [20]
reported an invertebrate succession on pig carcasses in aquatic
habitats in British Columbia, according to trophic adaptations
found in different taxa and available food sources.
This study establishes ecological categories by combining
strictly aquatic organisms found to be associated with the
submerged parts of the carcasses and terrestrial organisms
collected on the exposed parts during floating stages of decompo-
sition. A first group is composed of shredders and collectors, as well
as some predators in the lake. The shredders and collectors are
frequently found either in lake environments with an abundance of
macrophytes on the shores or in stream environments with gallery
forest, providing allochthonous organic matter in the form of
coarse particulate organic matter as a food source for shredders,
such as Tropisternus sp. (lake) and Hyallela sp. (stream). The
collectors, Chironomus sp. (collector–gatherer) and Simulium sp.
(collector–filterer) generally feed on fine particulate organic
171
matter, derived from the natural decay of organic matter and
from the biofilm formed on some substrates, such as carcasses,
which can be employed by these groups as secondary ecosystems
[37]. Maybe this fact favours the association of these groups with
decomposing bodies; however, these taxa are not related to any
specific stage of decomposition, implying that they will not be
good indicators of PMSI. However, they could be useful in
determining whether bodies have been moved, bearing in mind
that they are specifically found in waters with high primary
productivity.
In both habitats, during the early floating and floating decay
stages, new groups belonging to terrestrial environments appear,
such as predators represented by O. discicolle; necrophages, mainly
represented by C. nigribasis; and sarcosaprophages. These groups
colonised the exposed parts of the carcass due to floatation caused
by gases produced by bacterial activity within the gastric cavity
[17]. The necrophagous activity of C. nigribasis larvae, which eat the
soft tissues of the carcass, caused small holes to be made, allowing
gases to escape, leading to the onset of the next stage. It is possible
that these Calliphoridae are indicators of the early floating stage,
due to the carcass becoming available to colonisation by these
terrestrial insects. Groups associated with the submerged remains
stage were similar to those at the beginning of the decomposition
process (collectors and shredders in both habitats), suggesting that
the carcass becomes a food source when it emerges at the water’s
surface, whereas, when submerged, it represents a substrate for
anchorage and protection for strictly aquatic groups and not a food
source to collectors and shredders present at the final stage. In
support of the above, Sorg et al. [18] indicate that carcasses found
in aquatic environments comply with several ecological functions
(food source, refuge, provision of microhabitats and substrate).
3.2. Environmental factors
Biological decay is usually influenced by environmental factors.
In addition to biological mechanisms, the decomposition of a
carcass is determined by the physical, chemical and climatic
variables of its geographic location [30]. Of these factors,
environmental temperature probably has the most influence over
the decomposition process, given that it affects the time of
oviposition or colonisation, growth and survival of larvae, making
it the most important factor in estimating PMI in terrestrial
environments. It is to be expected that environmental temperature
will also have strong influence in aquatic systems, given that
organisms in this environment also respond to changes in
temperature. In a lake ecosystem, temperature is generally higher
than in stream waters, increasing the speed of decomposition [18].
However, the lower temperatures maintain approximately con-
stant levels of oxygen (favouring its solubility), which determines
the composition of the invertebrate community colonising a body
in a specific place, which, in turn, influences the rate and duration
of decomposition [20]. Variation in CO2 concentration in this study
was more evident in the lake during the late floatation stage,
maybe related to the high metabolic activity of intestinal bacteria
during this stage. Although variation in the concentration of CO2
could affect the abundance and diversity of a community, in this
case, it did not occur given that, during this time, the community
was mainly made up of necrophages and terrestrial predators
located on the exposed parts of the carcass. Conductivity showed a
similar behaviour to CO2 in the lake, whereas in the stream, these
variables were more constant over the sampling period and only
increased at the end of the study, coinciding with an increase in
rainfall, a factor which should be taken into account, given that
with the arrival of the rainy season, the water temperature varied,
as did CO2 and conductivity. Furthermore, the increased water
level and flow rate caused the early sinking of the remains in the
172 M. Barrios, M. Wolff / Forensic Science International 212 (2011) 164–172
lake during the bloated deterioration stage, and with this, the
disappearance of the terrestrial necrophages, mainly at larval
stages. This caused a decrease in trophic activity on the soft tissue,
whose decomposition and detachment was a result of the
mechanical action of the water current. This suggests that
wherever a body is recovered, it is fundamental to conduct a
complete inventory of environmental factors, as well as the routine
collections carried out by forensic entomologists to determine
either PMI or SPMI. In the tropics, ecological conditions are
markedly different within the same geographic region, making
extrapolation of succession tables and decomposition times more
difficult, and even more so, if trying to compare with temperate
regions.
This study represents a first approach to exploring the
succession of decomposers on carcasses in aquatic habitats to
estimate PMSI. However, given that carcasses were not replicated
and only placed in two bodies of water, it is essential to replicate
the data and extend the study to diverse aquatic ecosystems to
further develop the model.
4. Conclusion
PMSI for both carcasses was similar in the two habitats, each
showing six stages of decomposition, confirming that decomposi-
tion in freshwater ecosystems is different from that documented for
terrestrial environments. Benthic arthropods and neuston, with
trophic roles of collectors and shredders, are indicators of the
submersion stages, while C. nigribasis (necrophagous) and O.
discicolle (predator) are indicators of the floatation stages. Freshwa-
ter arthropods have the potential to be used to estimate PMSI, even
though several groups did not show trophic roles associated with
carcass decomposition. However, the latter groups could be used to
determine movements of bodies and the time of decomposition,
taking into account that they are specific to mesotrophic waters.
Water temperature, CO2, conductivity and rainfall are abiotic factors
which possibly affect the decomposition process by producing
different ecological conditions within the same region.
Acknowledgements
We are grateful to professor Emilio Realpe and the Zoological
and Ecological Aquatic Laboratory at the Universidad de Los Andes
(LAZOEA) for logistic support and to the Forensic Science Faculty of
the National Police Force, for their support in the field work.
References
[1] K.G.V. Smith, A Manual of Forensic Entomology, Trustees of the British Museum
(Natural History) and Cornell University Press, London, 1986, p. 205.
[2] G. Anderson, S.L. Van Laerhoven, Initial studies on insect succession on carrion in
Southwestern British Columbia, J. Forensic Sci. 41 (1996) 617–625.
[3] D.E. Gennard, Forensic Entomology—An Introduction, Wiley, 2007,, p. 224.
[4] M. Barreto, M.E. Burbano, P. Barreto, Flies (Calliphoridae, Muscidae) and Beetles
(Silphidae) from human cadavers in Cali, Colombia, Mem Inst Oswaldo Cruz 97 (1)
(2002) 137–138.
[5] J.R. Pujol-Luz, H. Marques, A. Ururahy-Rodrigues, J. Albertino Rafael, F.H.A.
Santana, L.C. Arantes, R. Constantino, A forensic entomology case from the
Amazon rain forest of Brazil, J. Forensic Sci. 51 (5) (2006) 1151–1153.
[6] J.R. Pujol-Luz, P.A. Costa Francez, A. Ururahy-Rodrigues, R. Constantino, The Black
Soldier-fly, Hermetia illucens (Diptera, Stratiomyidae), used to estimate the postmor-
tem interval in a case in Amapa’ State, Brazil, J. Forensic Sci. 53 (2) (2008) 476–478.
[7] L.M. Carvalho, P.J. Thyseen, A.X. Linhares, F.A. Palhares, A checklist of arthropods
associated with pig carrion and human corpes in Southeastern Brazil, Men inst
Oswaldo Cruz 95 (1) (2000) 135–138.
[8] L.M. Carvalho, A.X. Linhares, Seasonality of insect succession and pig carcass
descomposition in a natural forest area in Southeastern Brazil, J. Forensic Sci. 46
(3) (2001) 604–608.
[9] M. Wolff, A. Uribe, A. Ortı́z, P. Duque, A preliminary study of forensic entomology
in Medellı́n, Colombia, Forensic Sci. Int. 120 (1/2) (2001) 53–59.
[10] S. Pérez, P. Duque, M. Wolff, Successional behavior occurrence matrix of carrion-
associated arthropods in the Urban Area of Medellin, Colombia, J. Forensic Sci. 50
(2) (2005) 448–454.
[11] E. Martinez, P. Duque, M. Wolff, Succession pattern of carrion-feeding insects in
Paramo, Colombia, Forensic Sci. Int. 166 (2007) 182–189.
[12] J.A. Payne, A summer carrion study of the baby pig Sus scrofa Linnaeus, Ecology 46
(5) (1965) 592–602.
[13] J.A. Payne, E.W. King, Insect succession and decomposition of pig carcasses in
water, J. Georgia Entomol. Soc. 7 (1972) 153–162.
[14] N.H. Haskell, D.G. McShaffrey, D.A. Hawley, R.E. Williams, J.E. Pless, Use of aquatic
in determining submersion intervalo, J. Forensic Sci. 34 (1989) 622–632.
[15] D.A. Hawley, N.H. Haskell, D.G. Mcshaffrey, R.E. Williams, J.E. Pless, Identification
of a red fiber: Chironomidae larva, J. Forensic Sci. 34 (1989) 617–621.
[16] G.M. Vance, J.K. VanDyk, W.A. Rowley, A device for Sampling Aquatic Insects
associated with carrion in water, J. Forensic Sci. 40 (1995) 479–482.
[17] J.B. Keiper, E.G. Chapman, B.A. Foote, Midge larvae (Diptera: Chironomidae) as
indicators of postmortem submersion interval of carcasses in a woodland stream:
a preliminary report, J. Forensic. Sci. 42 (1997) 1074–1079.
[18] M.H. Sorg, J.H. Dearborn, E.J. Monahan, H.F. Ryan, K.G. Sweeney, E. David, Forensic
taphonomy in marine contexts, in: W.D. Haglund, M.H. Sorg (Eds.), Forensic
Taphonomy: The Postmortem Fate of Human Remains, CRC Press, Boca Raton,
1997, pp. 531–584.
[19] J.K. Tomberlin, P.H. Adler, Seasonal colonization and decomposition of rat carrion
in water and on land in an open field in South Carolina, J. Med. Entomol. 35 (1998)
704–709.
[20] N.R. Hobischack, Freshwater invertebrate succession and decompositional stud-
ies on carrion in British Columbia. Canadian Police Research Center, Technical
Report, Otawwa, ON, 1998, 54.
[21] N.R. Hobischak, G.S. Anderson, Freshwater-related death investigations in British
Columbia in 1995–1996. A review of coroners cases, Can. Soc. Forensic Sci. J.
(1999).
[22] J.B. Keiper, D.A. Casamatta, Benthic organisms as forensic indicators, North Am.
Benthol. Soc. 20 (2) (2001) 311–324.
[23] G.S. Anderson, N.R. Hobischak, Initial studies on insect succession on carrion on
British Columbia, Int. J. Legal Med. 108 (2004) 206–209.
[24] N.R. Hobischak, G.S. Anderson, Time of submergence using aquatic invertebrate
succession and decompositional changes in British Columbia, J. Forensic Sci. 47
(2002) 142–151.
[25] M. Wolff, A. Builes, G. Zapata, G. Morales, M. Benecke, Detection of parathion (O,O-
diethyl O-(4-nitrophenyl)phosphorothioate) by HPLC in insects of forensic im-
portance in Medellı́n, Colombia, Cases report Special Issue, Anil Aggrawal Internet
J. Forensic Med. Toxicol. (2004) 6–11. http://geradts.com/anil/ij/indexpapers.
html.
[26] M. Wolff, G. Zapata, G. Morales, M. Benecke, Detención y Cuantificación de Baygon
(Propoxur) en la sucesión de Insectos de Importancia Medico-Legal, Revista
Sociedad Colombiana de Entomologı́a 32 (2006) 159–164.
[27] N.A. Segura, W. Usaquen, M.C. Sanchez, L. Chuaire, F. Bello, Succession pattern of
cadaverous entomofauna in a semi-rural area of Bogotá, Colombia, Forensic Sci.
Int. 187 (2009) 66–72.
[28] Alcaldı́a de Facatativá, Plan de ordenamiento territorial, 2005, 64–66.
[29] R.W. Merritt, R.W. Cummins, An Introduction to the Aquatic Insects of North
America, 3rd ed., Kendall/Hunt Publishing Company, 1996, pp. 38–652.
[30] R.W. Merritt, J.R. Wallace, The role of aquatic insects in forensic investigations, in:
J. Byrd, J. Castner (Eds.), Forensic Entomology: The Utility of Arthropods in Legal
Investigations, CRC Press, Boca Raton, Florida, 2001 , p. 418 (Chapter 6,
pp. 178–215).
[31] M.S. Wipfli, J. Hudson, J. Caouette, Influence of salmon carcasses on stream
productivity: response of biofilm and benthic macroinvertebrates in Southeast-
ern Alaska, USA, Can. J. Fisheries Aquat. Sci. 55 (1998) 1503–1511.
[32] J.H. Byrd, J.L. Castner, Insects of forensic importance, in: J. Byrd, J. Castner (Eds.),
Forensic Entomology: The Utility of Arthropods in Legal Investigations, CRC Press,
Boca Raton, FL, 2001, p. 418 (Chapter 2, pp. 44–79).
[33] W.R. Rodrı́guez, Decomposition of buried and submerged bodies, in: Haglund,
Sorg (Eds.), Forensic Taphonomy: The Post-mortem Fate of Human Remains, CRC
Press LLC, Boca Raton, FL, 1997, pp. 459–468.
[34] B. Knight, Simpson’s Forensic Medicine, 11th ed., Arnold, London, 1997,
pp. 115–118.
[35] G.A. Cole, Manual de Limnologı́a, Editorial Hemisferio Sur S.A. Buenos Aires,
Argentina, 1988, p. 405.
[36] R.W. Cummins, Structure and function of stream ecology, Bioscience 24 (1974)
631–641.
[37] J.V. Ward, Aquatic Insect Ecology. 1. Biology and Habitat, John Wiley & Sons, Inc.,
New York, 1992, p. 384.