ALBERT

Description: In this experiment, we assessed the different transmission steps from the first to the second intermediate host: i) cercarial emergence from periwinkles, ii) cercarial activity and survival after emergence, iii) cercarial infectivity in mussels, and iv) susceptibility of mussels to cercarial infection. For iii) cercariae were treated but not the mussels, whereas for iv) mussels were treated but not the cercariae. The experiment was run in August-September 2017 in the climate chambers of GEOMAR in Kiel. All experiments were conducted using temperature and salinity (fully crossed) as well as time (only for cercarial output) as fixed factors, and periwinkle/mussel identity nested within water bath as random factor. Temperature levels applied were 19 and 23°C. Salinity levels applied were 13, 16 and 19.

Description: Cercarial emergence was assessed by counting all cercariae that emerged from single periwinkles exposed to the different treatments. Periwinkles were collected in Årøsund, Denmark and screened in the laboratory to verify the infection status. To measure cercarial emergence, periwinkles were individually transferred from the jars into 50 mL Plexiglas beakers, one day after the 7-days acclimation (sampling 1), one week after the start of exposure to the treatments (sampling 2), and two weeks after the start of exposure to the treatments (sampling 3). Each incubation lasted eight hours. At the end of the incubation period, the water from each beaker was transferred into 50 mL Falcon tubes. In addition, the beakers were immediately washed with 5 mL ethanol (99%), and the solution was also added to the Falcon tube to ensure the collection of all cercariae and their preservation for later counting.

Description: For cercarial activity and survival infected periwinkles were acclimated for three weeks under the different treatments (n=10 individuals per treatment) and incubated to allow for cercarial release. At the start of the experiment, approximately 45 cercariae per treatment were added individually to wells of three 96-well plates The 96-well plates were then exposed to the different temperature treatments placing them in the six thermobaths (three 96-well plates per treatment combination). Activity and survival were then assessed under a stereomicroscope after 4, 6, 9, 18, 27, and 45 hours. Cercariae were considered fully active when they were constantly swirling around (category: fully active). When laying at the bottom and not reacting, these were considered dead (category: dead), when still reacting, even if slowly, these were still considered alive (category: alive). This last category included "fully active".

Description: The infectivity of H. elongata cercariae was investigated by counting metacercariae in M. edulis, after a standardized exposure to cercariae acclimated at different temperatures and salinities. Mussels were collected two days prior to the start of the assays from the Kieler Meeresfarm. Then, A total of 72 mussels (n=12 for each treatment combination) were individually placed in 50 mL beakers at the respective temperatures and salinities, and exposed to a standardized number of cercariae released from the all periwinkles from the different treatment combinations. In each water bath, four beakers per salinity combination were placed. The 16-23°C treatment only five mussels where infected with 20 cercariae each due to the low number of cercariae obtained from such pre-treated periwinkles. The 13-23°C treatment was excluded from the analyses due to extremely low numbers of cercariae released under this particular treatment combination. The beakers with mussels were then incubated for 24 hours. After the incubation, each mussel was collected, dissected, and H. elongata metacercariae were identified and counted under a stereomicroscope. The effects of temperature and salinity on the susceptibility of M. edulis to infection by H. elongata cercariae were assessed by acclimating mussels to the above-mentioned treatment combinations (salinity x temperature) for one week, before these were exposed to non-acclimated cercariae. The exposure to cercariae of each mussel (n=12 mussels per treatment) was realized as described for Exp. III. Beakers with mussels were incubated at the different treatments for 24 hours. Mussels were then dissected and the number of metacercariae in each mussel (squeezed between two glass plates) was counted under a stereomicroscope.

Description: In this study, we examined how chemical cues from a predatory marine crab affect the transmission of a parasitic trematode from its first (periwinkle) to its second (mussel) intermediate host. We collected the data in a laboratory experiment. Here, snails (Littorina littorea) infected with a parasite (Himasthla elongata) were kept in two different treatments (with predation risk and control). Subsequently, the excreted cercariae were collected as data. The experiments were conducted at the Wadden Sea Station of the Alfred Wegener Institute in List, Sylt, Germany. We sampled the snails at the the Danish coast of the Baltic Sea (Jütland, Arosund; 55*15'45.8'N 9*42'39.2'E). Snails had a shell height of 14-18mm corresponding to an age of two years. Infection status were screened at the laboratory. The crabs for the predation cue were sampled at the Oddewatt, List Sylt (German, Wadden Sea). Only male crabs with a size og 20-30mm catapace width were sampled. The blue mussels were sampled at the west coast of Sylt (Wenningstedt beach) were trematode infection do not occur naturally (confirmed by screening 50 mussels). Mussel shell length wars 25-30mm.

Description: In this study, we examined how chemical cues from a predatory marine crab affect the transmission of a parasitic trematode from its first (periwinkle) to its second (mussel) intermediate host. We collected the data in a laboratory experiment. Here, snails (Littorina littorea) infected with a parasite (Himasthla elongata) were kept in two different treatments (with predation risk and control). Subsequently, the excreted cercariae were collected as data. The experiments were conducted at the Wadden Sea Station of the Alfred Wegener Institute in List, Sylt, Germany. We sampled the snails at the the Danish coast of the Baltic Sea (Jütland, Arosund; 55*15'45.8'N 9*42'39.2'E). Snails had a shell height of 14-18mm corresponding to an age of two years. Infection status were screened at the laboratory. The crabs for the predation cue were sampled at the Oddewatt, List Sylt (German, Wadden Sea). Only male crabs with a size og 20-30mm catapace width were sampled. The blue mussels were sampled at the west coast of Sylt (Wenningstedt beach) were trematode infection do not occur naturally (confirmed by screening 50 mussels).

Description: In this study, we examined how chemical cues from a predatory marine crab affect the transmission of a parasitic trematode from its first (periwinkle) to its second (mussel) intermediate host. We collected the data in a laboratory experiment. Here, snails (Littorina littorea) infected with a parasite (Himasthla elongata) were kept in two different treatments (with predation risk and control). Subsequently, the excreted cercariae were collected as data. The experiments were conducted at the Wadden Sea Station of the Alfred Wegener Institute in List, Sylt, Germany. We sampled the snails at the the Danish coast of the Baltic Sea (Jütland, Arosund; 55*15'45.8'N 9*42'39.2'E). Snails had a shell height of 14-18mm corresponding to an age of two years. Infection status were screened at the laboratory. The crabs for the predation cue were sampled at the Oddewatt, List Sylt (German, Wadden Sea). Only male crabs with a size og 20-30mm catapace width were sampled. The blue mussels were sampled at the west coast of Sylt (Wenningstedt beach) were trematode infection do not occur naturally (confirmed by screening 50 mussels). Mussel shell length wars 25-30mm.

Description: In this study, we examined how chemical cues from a predatory marine crab affect the transmission of a parasitic trematode from its first (periwinkle) to its second (mussel) intermediate host. We collected the data in a laboratory experiment. Here, snails (Littorina littorea) infected with a parasite (Himasthla elongata) were kept in two different treatments (with predation risk and control). Subsequently, the excreted cercariae were collected as data. The experiments were conducted at the Wadden Sea Station of the Alfred Wegener Institute in List, Sylt, Germany. We sampled the snails at the the Danish coast of the Baltic Sea (Jütland, Arosund; 55*15'45.8'N 9*42'39.2'E). Snails had a shell height of 14-18mm corresponding to an age of two years. Infection status were screened at the laboratory. The crabs for the predation cue were sampled at the Oddewatt, List Sylt (German, Wadden Sea). Only male crabs with a size og 20-30mm catapace width were sampled. The blue mussels were sampled at the west coast of Sylt (Wenningstedt beach) were trematode infection do not occur naturally (confirmed by screening 50 mussels). Mussel shell length wars 25-30mm. Sampling of the experimental organisms: Mytilus edulis (latitude:54.937.600, longitude: 8.312.915); Littorina littorea (latitude55.262.722:longitude:9.710.889 ) and Hemigrapsus takanoi (latitude:55.028.713longitude:8.434.260 ).