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. 2021 Nov 20;12(11):1043. doi: 10.3390/insects12111043

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Setups for experiments in the pressure vessel IPOCAMP. (a) The pressure vessel IPOCAMP (internal diameter 20 cm, height 60 cm [36]); (b) experimental setup for grooming observations. Arrows indicate the inlet and outlet of circulating seawater. This device also used for chemical stimuli experiments, see (c), is a recently upgraded version of the lid of the vessel IPOCAMP. The new lid comprises a large viewport and consequently allows direct observation as well as video recording with a high definition camera (ca, AG-HCK10G HD camera head, AG-HMR10 portable recorder, Panasonic). Three shrimp were placed in a PVC cage closed at the top with a transparent polyethylene lid. The behavior of the shrimp was recorded throughout the experiment. Pictures (b1–b3) are views of the following animals in the IPOCAMP aquarium during grooming observations: (b1) R. exoculata females and male (marked with a black line); (b2) R. exoculata juveniles; (b3) Palaemon serratus juveniles. Body length of the observed shrimp is about 4–5 cm for R. exoculata and P. serratus juveniles, and about 2 cm for R. exoculata juveniles. (c) Experimental setup for chemical stimuli experiments. Arrows indicate the inlet and outlet of circulating seawater. The seawater inlet pipe passes through a thermostatically controlled bath. The lid is equipped with an isobaric line (i) that allows the introduction of small elements (e.g., food, stimulus) without disrupting the pressure inside the aquarium. During the experiment, two control gels (grey bars) and one stimulus gel (black bar) were introduced into the tank through the isobaric line at 45 -min intervals. Picture (c1) shows Mirocaris fortunata in the IPOCAMP aquarium during this experiment. Specimens lie on the plastic bottom of the tank with holes for water entrance, two stainless steel tubes that contain gels are visible. (d) Experimental setup for temperature and sulfide pulse experiments. Three sapphire viewports in the pressure vessel lid allow the insertion of an endoscope (e) (Fort, Dourdan, France) and two optical -fiber light -guides for the behavioral observation. The experiments are recorded by a CCD camera (JVC, TK-C1380) and a DVD recorder (DVO-1000MD, Sony). A diffuser system, consisting of a plastic cap with holes, is placed over the seawater inlet hole. Warm -temperature pulses were obtained by immersing a heat exchanger (HE) located on the water inlet line in a temperature -controlled water bath that had been preset to the desired pulse temperature. Two Pt-100 autonomous temperature loggers (S2T6000D, NKE Instruments) are positioned in the upstream water flow (T1) and on the plastic plate at the bottom of the tank several centimeters (>5 cm) from the hot water diffuser (T2). For the sulfide pulse experiments, the HE2 and temperature loggers were removed, and the sulfide solutions were injected into the seawater inlet pipe. Picture (d1) shows Rimicaris exoculata in IPOCAMP during a temperature pulse experiment at 25 °C. Specimens gathered around the warm water diffuser (marked with a dark circle).