Homarus Gammarus Research Articles

RETRACTED: Improving tilapia (Oreochromis mossambicus) resistance to streptococcal disease by improving the gut biome through administration of the microorganisms Bacillus subtilis and Enterococcus faecalis

This article has been retracted: please see Elsevier Policy on Article Withdrawal ( http://www.elsevier.com/locate/withdrawalpolicy ). This article has been retracted at the request of the Editor. The authors have plagiarized part of a paper that had already appeared in Holt, C.C., van der Giezen, M., Daniels, C.L. et al. Spatial and temporal axes impact ecology of the gut microbiome in juvenile European lobster (Homarus gammarus). ISME J 14, 531–543 (2020). https://doi.org/10.1038/s41396-019-0546-1 One of the conditions of submission of a paper for publication is that authors declare explicitly that their work is original and has not appeared in a publication elsewhere. Re-use of any data should be appropriately cited. As such this article represents a severe abuse of the scientific publishing system. The scientific community takes a very strong view on this matter and apologies are offered to readers of the journal that this was not detected during the submission process.

Replacement of Antarctic krill (Euphausia superba) by extruded feeds with different proximate compositions: effects on growth, nutritional condition and digestive capacity of juvenile European lobsters (Homarus gammarus, L.).

Extruded feeds are widely used for major aquatic animal production, particularly for finfish. However, the transition from fresh/frozen to extruded/pelleted feeds remains a major obstacle to progressing sustainable farming of European lobster (Homarus gammarus). The aim of the present study was to investigate the effects of using extruded feeds with different protein levels and lipid/carbohydrate ratios on growth, feed utilisation, nucleic acid derived indices (sRD) and digestive enzymatic activity of H. gammarus juveniles. Six extruded feeds were formulated to contain two protein levels (400 and 500 g/kg), with three lipid/carbohydrate ratios (LOW - 1:3; MEDium - 1:2; HIGH - 1:1). The extruded feeds were tested against Antarctic krill (Euphausia superba) used as control (CTRL). Overall, the CTRL and 500MED feed supported the highest growth and nutritional condition estimated by means of sRD, while the poorest results were observed for the 400HIGH and 400MED groups. The FCR was significantly lower in the CTRL than all extruded feeds, among which the most efficient, i.e., lower FCR, was the 500MED. The highest activity of trypsin and amylase in lobsters fed the 400MED and 400HIGH feeds points to the activation of a mechanism to maximise nutrients assimilation. The highest lipase activity observed for the 500LOW and 500MED groups indicates a higher capacity to metabolise and store lipids. Overall, the results suggest that the 500MED feed (500 g/kg protein, 237 g/kg carbohydrates and 119 g/kg lipids) is a suitable extruded feed candidate to replace Antarctic krill, commonly used to grow lobster juveniles.

Marine lobsters in ancient Greek and Latin

This paper describes the Ancient Greek and Latin terminology for marine lobsters, especially for ‘European lobster, Homarus gammarus L.’, ‘common spiny Lobster, Palinurus elephas Fabricius’ and ‘No...

Population Genetic Structure and Connectivity of the European Lobster Homarus gammarus in the Adriatic and Mediterranean Seas.

Highly selective fishing has the potential to permanently change the characteristics within a population and could drive the decline of genetic diversity. European lobster is an intensively fished crustacean species in the Adriatic Sea which reaches high market value. Since knowledge of population structure and dynamics is important for effective fisheries management, in this study, we used 14 neutral microsatellites loci and partial mitochondrial COI region sequencing to explore population connectivity and genetic structure by comparing samples from the Adriatic Sea and the adjacent basins of the Mediterranean Sea. The obtained results suggest that neutral genetic diversity has not been significantly affected by decrease in population size due to overfishing, habitat degradation and other anthropogenic activities. Global genetic differentiation across all populations was low (FST = 0.0062). Populations from the Adriatic Sea were panmictic, while genetic differentiation was found among populations from different Mediterranean basins. Observed gene flow for European lobster suggest that populations in the north eastern Adriatic act as a source for surrounding areas, emphasizing the need to protect these populations by establishing interconnected MPAs that will be beneficial for both fisheries and conservation management.

Effects of short-term and continuous exposure to reduced salinities on the biochemical composition of larval lobster, Homarus gammarus

In coastal areas with estuarine influence, exposure to hypo-osmotic conditions may affect larval survival, development and growth. Most knowledge about effects of reduced salinity on coastal organisms is based on keeping individuals under constant conditions in the laboratory. By contrast, little is known about the effects of more realistic situations where organisms are exposed to low salinity over short time scales. Such environmental short-term fluctuations are expected to increase due to climate change. Here, we experimentally evaluated the sublethal effects of both short-term and continuous exposure to moderately reduced salinities (salinity 20 and 25; compared to seawater, salinity 32) in larvae of European lobster Homarus gammarus. Total body dry mass and biochemical composition (measured as: protein and lipid contents) were measured as response variables in Mysis stages I to III. Short-term effects of low salinity were quantified in a group of larvae kept in seawater from hatching until the time of transfer to the test salinities. After ca. 40 % of each moult cycle in seawater (determined in preliminary experiments for Mysis I, II and III), larvae were assigned to a seawater control or reduced salinities lasting for 16 h (i.e. until ca. 50 % of the time spent within the moulting cycle). Effects of continuous exposure to low salinity were quantified when larvae were exposed to the different salinities from hatching, until they reached ca. 50 % of the successive moulting stage. Surprisingly, in the Mysis II and III stages, short-term exposure to low salinity had much stronger effects on accumulation of reserves than the continuous exposure. Such effects were manifested mostly as limited accumulation, or even losses, in the lipid content as compared to reductions in the amount of protein accumulated. The most sensitive stage to exposure to low salinity was the Mysis III; by contrast in Mysis I such effects were relative weak (not always significant). Chronic exposure to low salinity also led to an increase in developmental time especially at the advanced stages. Our results highlight the importance of quantifying effects of environmental fluctuations at different time scales in order to better understand how organisms cope with realistic environmental change in the coastal zones. For H. gammarus, our results suggest that larvae respond adaptively to low salinity by maintaining protein levels at expenses of reductions in lipid accumulation and by extending the developmental time, but the capacity to elicit a fully compensatory response varies ontogenetically.

From the Adriatic to Northern Norway—geographic differences in moult increment and moult probability of the European lobster (Homarus gammarus), across the natural range

Abstract This study collated existing data on lobster moult increment from studies across the range of Homarus gammarus, together with new tagging data from Orkney, United Kingdom. Generalized additive models were used to investigate geographical differences in absolute moult increment and moult probability. Absolute moult increment was seen to differ significantly between regions and between sexes and showed a non-linear relationship with pre-moult size. Smaller absolute moult increments were observed at southern and northern latitudes with larger increments observed in the centre of the species range. Temperature was identified as a significant factor explaining differences in absolute moult increment between regions, consistent with there being a thermal optimum for growth for in H. gammarus. Moult and double moult probabilities decline with pre-moult size, with greatest probability of moulting occurring around day 250. Probability of double moulting increased with mean annual sea surface temperature. Regionally variable growth patterns, and localized adaptation to abiotic variables such as temperature, should therefore be taken into account when defining lobster stock assessment and fishery management areas.

European lobsters utilise Atlantic salmon wastes in coastal integrated multi-trophic aquaculture systems

In this study, we investigated if juvenile European lobstersHomarus gammaruswould eat waste from Atlantic salmonSalmo salarcages in a coastal integrated multi-trophic aquaculture (IMTA) setup and if there were any impacts on growth. Trophic interactions between salmon and lobsters were assessed using δ15N and δ13C stable isotope analysis and fatty acid profiling from fish feed as indicators of nutrient flow. Analysis revealed that lobsters directly utilised particulate waste from salmon production, as levels of indicator fatty acids from salmon feed were significantly higher in lobster tissues near the fish cages compared to the control site. Route of uptake may have been direct consumption of waste feed or faecal material or indirectly through fouling organisms. Stable isotope analysis did not indicate nutrient transfer to lobsters, suggesting that the duration of the study and/or the amount of waste consumed was not sufficient for stable isotope analysis. Lobsters grew significantly over the trial period at both sites, but there was no significant difference in lobster growth between the sites. Our results show a trophic relationship between salmon and lobsters within this IMTA system, with no apparent advantage or disadvantage to growth.

Aristotle\u2019s lobster: the image in the text

The Anatomai, a lost work written by Aristotle, must have contained a collection of various drawings and figures of species as well as their organs. In his texts (mainly the Historia animalium), Aristotle is often referring to the drawings after the description of species. Our study applies the method of the comparative view (‘Vergleichendes Sehen’) to provide an access to and reconstruction of Aristotle’s lost illustrations based on his textual descriptions. As an example, we chose the treatment of the European lobster (Homarus gammarus L., 1758) in the Aristotelian corpus as a case study. First, we analyse the etymology of the Greek term astakós referring to the lobster and provide an overview on the putative synonyms. Second, we confront the textual basis of the description with several questions concerning the degree of abstraction, the relation between text and image, and the spatial orientation of the image. Finally, we present a step-by-step reconstruction of Aristotle’s illustrations of the lobster based on the various passages dealing with its anatomy in the text of the Historia animalium. The problems which arise by a confrontation of the textual basis with hypothetical images are discussed at a more general level. We conclude that this kind of a text-based image reconstruction is only possible if the object described by Aristotle is unambiguously identifiable and still visually accessible.

Bioavailability of Microplastics to Marine Zooplankton: Effect of Shape and Infochemicals.

The underlying mechanisms that influence microplastic ingestion in marine zooplankton remain poorly understood. Here, we investigate how microplastics of a variety of shapes (bead, fiber, and fragment), in combination with the algal-derived infochemicals dimethyl sulfide (DMS) and dimethylsulfoniopropionate (DMSP), affect the ingestion rate of microplastics in three species of zooplankton, the copepods Calanus helgolandicus and Acartia tonsa and larvae of the European lobster Homarus gammarus. We show that shape affects microplastic bioavailability to different species of zooplankton, with each species ingesting significantly more of a certain shape: C. helgolandicus-fragments (P < 0.05); A. tonsa-fibers (P < 0.01); H. gammarus larvae-beads (P < 0.05). Thus, different feeding strategies between species may affect shape selectivity. Our results also showed significantly increased ingestion rates by C. helgolandicus on all microplastics that were infused with DMS (P < 0.01) and by H. gammarus larvae and A. tonsa on DMS-infused fibers and fragments (P < 0.05). By using a range of more environmentally relevant microplastics, our findings highlight how the feeding strategies of different zooplankton species may influence their susceptibility to microplastic ingestion. Furthermore, our novel study suggests that species reliant on chemosensory cues to locate their prey may be at an increased risk of ingesting aged microplastics in the marine environment.

A novel nudivirus infecting the invasive demon shrimp Dikerogammarus haemobaphes (Amphipoda)

The Nudiviridae are a family of large double-stranded DNA viruses that infects the cells of the gut in invertebrates, including insects and crustaceans. The phylogenetic range of the family has recently been enhanced via the description of viruses infecting penaeid shrimp, crangonid shrimp, homarid lobsters and portunid crabs. Here we extend this by presenting the genome of another nudivirus infecting the amphipod Dikerogammarus haemobaphes. The virus, which infects cells of the host hepatopancreas, has a circular genome of 119,754 bp in length, and encodes a predicted 106 open reading frames. This novel virus encodes all the conserved nudiviral genes (sharing 57 gene homologues with other crustacean-infecting nudiviruses) but appears to lack the p6.9 gene. Phylogenetic analysis revealed that this virus branches before the other crustacean-infecting nudiviruses and shares low levels of gene/protein similarity to the Gammanudivirus genus. Comparison of gene synteny from known crustacean-infecting nudiviruses reveals conservation between Homarus gammarus nudivirus and Penaeus monodon nudivirus; however, three genomic rearrangements in this novel amphipod virus appear to break the gene synteny between this and the ones infecting lobsters and penaeid shrimp. We explore the evolutionary history and systematics of this novel virus, suggesting that it be included in the novel Epsilonnudivirus genus (Nudiviridae).

Renewable energy homes for marine life: Habitat potential of a tidal energy project for benthic megafauna

An increasing number of offshore structures are being deployed worldwide to meet the growing demand for renewable energy. Besides energy production, these structures can also provide new artificial habitats to a diversity of fish and crustacean species. This study characterises how concrete mattresses that stabilise the submarine power cable of a tidal energy test site can increase habitat capacity for benthic megafauna. A five-year monitoring, which relied on both visual counts and video-based surveys by divers, revealed that these mattresses provide a suitable habitat for 5 taxa of large crustaceans and fish. In particular, two commercially valuable species, i.e. the edible crab Cancer pagurus and the European lobster Homarus gammarus, showed a constant occupancy of these artificial habitats throughout the course of the project. The shape and the number of shelters available below individual mattresses largely determine potential for colonisation by mobile megafauna. Local physical characteristics of the implantation site (e.g. substratum type, topography, exposition to current etc.) significantly impact amount and type of shelters provided by the concrete mattresses. Thus, to characterise habitat potential of artificial structures, it is not only essential to consider (i) the design of the structures, but also to (ii) account for their interactions with local environmental conditions when deployed on the seafloor.

Short-term exposure to hydrogen peroxide induces mortality and alters exploratory behaviour of European lobster (Homarus gammarus)

Bath treatment chemotherapeutants, used to control sea lice infestations in the salmonid aquaculture industry, are released directly into the marine environment around fish farms and pose a serious risk to non-target species, particularly crustaceans. Hydrogen peroxide (H2O2) is the most frequently used bath treatment chemotherapeutant on Norwegian fish farms, however, limited information is available on its toxicity to European lobsters (Homarus gammarus), a commercially important species at risk of exposure due to its distribution overlapping with salmon farm locations. The aim of this study was to investigate the lethal effects of H2O2 on pelagic (stage I-IV) larvae/post-larvae and its sub-lethal effects on the benthic stage V H. gammarus. To assess the lethal effects of H2O2, we carried out a series of 1 h toxicity tests and assessed mortality after a 24 h post-exposure period. Exposure to H2O2 was toxic to all pelagic larval stages tested, with estimated median lethal concentrations (LC50) of 177, 404, 665 and 737 mg/L for stage I, II, III and IV, respectively. These concentrations represent approximately 10, 23, 40 and 43%, of the recommended H2O2 concentrations used for delousing salmon on Norwegian fish farms, respectively. To assess the sub-lethal effects of H2O2 on H. gammarus, stage V juveniles were exposed to H2O2 at concentrations of 85, 170 and 510 mg/L for 1 h and shelter-seeking behaviour and mobility endpoints were assessed. Numerous behavioural parameters including distance travelled to shelter, time to locate shelter and the number of shelter inspections, were negatively affected in lobsters exposed to H2O2 when assessed immediately after the exposure period. However, no differences between control and exposed lobsters were detected after a 24 h post-exposure period. Our results demonstrate that short term exposures to H2O2 are lethal to pelagic H. gammarus life stages and can negatively affect the shelter seeking behaviour of benthic life stages, though these behavioural changes may be short-lived.

Abundance and size of European lobsters (Homarus gammarus) and brown crabs (Cancer pagurus) inside and outside the Kåvra lobster reserve (west coast of Sweden)

Abstract The common method of using lobster-pot catch data for investigating relative abundance, sex and size distribution has serious disadvantages. This study estimates relative abundance and size of the European lobster Homarus gammarus and the brown crab Cancer pagurus using scuba diving techniques. The study areas were the Kåvra lobster reserve (Kåvra) on the Swedish west coast and three very different nearby areas where fishing for crustaceans is allowed: Gullmarsfjorden; the archipelago; and the offshore area. A total of 167 lobsters and 337 brown crabs were observed during 33 scuba dives (each 30 minutes long) in 2018-2019. The estimated mean abundance of lobsters was three to fifteen times as high at Kåvra in comparison with the other three areas (all exact showing that the statistical populations were distinct in comparison with Kåvra; Wilcoxon-Mann-Whitney Test). Dive sites in the outer part of the offshore area had the lowest abundance of lobsters although they had seemingly good lobster habitats and low fishing intensity. Large lobsters with a carapace length of ⩾15 cm were found only at Kåvra where they made up 24% of the lobsters. The proportion of large lobsters inside Kåvra continues to increase after 30 years of protection. Together, this emphasizes the impact of fishing on lobster abundance and size distribution, and indicates that limited recruitment and migration might possibly affect offshore lobster “sub populations”. Kåvra was the only area where the abundance of lobsters exceeded the abundance of brown crabs (). However, the abundance of brown crabs at Kåvra was as low as in Gullmarsfjorden () where fishing for crabs is allowed. Possible complex lobster/brown crab interactions together with other factors that might explain the low abundance of the protected brown crab at Kåvra, need to be investigated further.

Preliminary Results on the Growth of Larval European Lobster (Homarus gammarus (Linneaus, 1758)) in Turkey

Sea lobsters are among the most valuable seafood traded commodities. Since its production with fishing has decreased every year in the world, many types of cultural studies have been carried out and success has been achieved. The aim of this research is to investigate the survival of European lobster (Homarus gammarus) larvae in Turkey and to determine the nature of the direction of growth opportunities. This research is the first study in Turkey investigating the nature of the direction of growth opportunities and the survival of European lobster larvae. Larval release, larvae feeding and survival possibilities were investigated on two egg-bearing lobsters obtained from Çanakkale coasts. Both broodstock larvae were able to survive until the post larval stage. The larvae of the first mature lobster reached 10.857 mm total length and 0.025 g live weight after approximately 30 days. The larvae of the second mature lobster reached 26.9 mm total length and 0.502 g live weight after 33 days. A significant difference was found in the larvae of two mature lobsters at the end of the experiment according to their initial dimensions in both length and weight (p &amp;lt;0.05). In addition, it was determined that the growth was higher due to the higher temperature in the larvae of the second mature lobster.

A review of American lobster (Homarus americanus) records around the British Isles: 2012 to 2018

Although native to eastern Canada and the USA, American lobsters (Homarus americanus) have been reported from British coastal waters in recent years. The introduction of most have been as escapees from holding facilities or releases by members of the public for animal welfare or religious reasons. Life-history traits of H. americanus put them at a potential competitive advantage over and hybridisation with the European lobster (Homarus gammarus), the 5th most valuable seafood to the UK. While yearly reported numbers of H. americanus found in British waters have been low, a mass release of 361 H. americanus into the English Channel off Brighton, southern England, took place in June 2015. This review identified 162 records of H. americanus in British waters between 2012 and 2018, with specimens caught mainly in or around Brighton, although additional H. americanus released may have gone undetected. While this evidence suggests a growing potential for establishment of the species, further information is provided evidencing hybridisation between released H. americanus and native H. gammarus via genetic analysis. This review highlights the need for a more robust means of detecting H. americanus and other non-native species in British waters.

Marine protected areas rescue a sexually selected trait in European lobster.

Marine protected areas (MPAs) are increasingly implemented worldwide to maintain and restore depleted populations. However, despite our knowledge on the myriad of positive responses to protection, there are few empirical studies on the ability to conserve species’ mating patterns and secondary sexual traits. In male European lobsters (Homarus gammarus), the size of claws relative to body size correlates positively with male mating success and is presumably under sexual selection. At the same time, an intensive trap fishery exerts selection against large claws in males. MPAs could therefore be expected to resolve these conflicting selective pressures and preserve males with large claws. We explored this hypothesis by contrasting claw size of males and females in three pairs of MPAs and nearby fished areas in southern Norway. By finding that male lobsters have up to 8% larger claws inside MPAs compared to similarly sized males in fished areas, our study provides evidence that MPAs rescue a secondary sexual trait. Recovery from harvest selection acting on claws is the most likely explanation; however, the higher abundance of lobster inside MPAs does not rule out a plastic response on claw size due to increased competition. Regardless of the underlying cause, our study demonstrates (a) the value of protected areas as a management tool for mitigating fisheries‐induced evolution and (b) that MPAs help maintaining the scope for sexual selection in populations with vulnerable life histories and complex mating system.

Crossing the pond: genetic assignment detects lobster hybridisation

American lobsters (Homarus americanus) imported live into Europe as a seafood commodity have occasionally been released or escaped into the wild, within the range of an allopatric congener, the European lobster (H. gammarus). In addition to disease and competition, introduced lobsters threaten native populations through hybridisation, but morphological discriminants used for species identification are unable to discern hybrids, so molecular methods are required. We tested an array of 79 single nucleotide polymorphisms (SNPs) for their utility to distinguish 1,308 H. gammarus from 38 H. americanus and 30 hybrid offspring from an American female captured in Sweden. These loci provide powerful species assignment in Homarus, enabling the robust identification of hybrid and American individuals among a survey of European stock. Moreover, a subset panel of the 12 most powerful SNPs is sufficient to separate the two pure species, even when tissues have been cooked, and can detect the introduced component of hybrids. We conclude that these SNP loci can unambiguously identify hybrid lobsters that may be undetectable via basic morphology, and offer a valuable tool to investigate the prevalence of cryptic hybridisation in the wild. Such investigations are required to properly evaluate the potential for introgression of alien genes into European lobster populations.

The impact of anti-sea lice pesticides, azamethiphos and deltamethrin, on European lobster (Homarus gammarus) larvae in the Norwegian marine environment

Anti-sea lice pesticides, used in the salmonid aquaculture industry, are a growing environmental concern due to their potential to adversely affect non-target crustaceans. Azamethiphos and deltamethrin are two bath treatment pesticides used on salmon farms in Norway, however, limited information is available on their impact on European lobster (Homarus gammarus) larvae in the Norwegian marine environment. Here, we firstly report the lethal (LC50) and effective (EC50) concentrations of azamethiphos and deltamethrin for stage I and stage II larvae, following 1-h exposures. Using a hydrodynamic model, we also modelled the dispersal of both compounds into the marine environment around selected Norwegian farms and mapped the potential impact zones (areas that experience LC50 and EC50 concentrations) around each farm. Our data shows that azamethiphos and deltamethrin are acutely toxic to both larval stages, with LC50 and EC50 values below the recommended treatment concentrations. We also show that the azamethiphos impact zones around farms were relatively small (mean area of 0.04–0.2 km2), however deltamethrin impact zones covered much larger areas (mean area of 21.1–39.0 km2). These findings suggest that deltamethrin poses a significant risk to European lobster in the Norwegian marine environment while the impact of azamethiphos may be less severe.

Historical translocations and stocking alter the genetic structure of a Mediterranean lobster fishery.

Stocking is often used to supplement wild populations that are overexploited or have collapsed, yet it is unclear how this affects the genetic diversity of marine invertebrate populations. During the 1970s, a lobster stock enhancement program was carried out around the island of Corsica in the Mediterranean using individuals translocated from the Atlantic coast of France. This included the release of thousands of hatchery‐reared postlarval lobsters and several adult individuals, but no monitoring plan was established to assess whether these animals survived and recruited to the population. In this study, we sampled European lobster (Homarus gammarus) individuals caught around Corsica and tested whether they showed Atlantic ancestry. Due to a natural marked phylogeographic break between Atlantic and Mediterranean lobsters, we hypothesized that lobsters with dominant (>0.50) Atlantic ancestry were descended from historical stocking releases. Twenty Corsican lobsters were genotyped at 79 single nucleotide polymorphisms, and assignment analysis showed that the majority (13) had dominant Atlantic ancestry. This suggests that the hatchery stocking program carried out in Corsica during the 1970s, using individuals translocated from the Atlantic coast of France, has likely augmented local recruitment but at a cost of altering the genetic structure of the Corsican lobster population.

Water Vapor Permeability of Chitosan/Zeolite Composite Films as Affected by Biopolymer and Zeolite Microparticle Concentrations

This work evaluated the effect of chitosan and zeolite microparticle concentrations on the water vapor permeability of composite films. In addition, others chemical, thermal, and physical properties of selected films were determined. Chitosan/zeolite composite films were prepared via the solution casting method using chitosans from chitin from common lobster (Panulirus argus) (77 and 275 kDa and ~ 75% of deacetylation degree) and natural zeolite (O = 35–45 µm). Chitosan concentration (1.5–2.0% w/v), zeolite microparticle concentration (0.0–7.0% w/w of chitosan), and molecular weight (MW) of chitosan were selected as factors through an IV optimal response surface design for the best variant of film based on the water vapor permeability (WVP). The WVP increased with an increase in the chitosan and zeolite concentration as well as the MW, being the film obtained from 1.5% (w/v) of chitosan (77 kDa) and 3.5% (w/w of chitosan) of zeolite, the film with the lower WVP. The increase in the chitosan concentration and zeolite addition at 3.5% (w/w of chitosan) decreased the elongation at break, while increased the tensile strength of the films. Water solubility increased with the polymer concentration, while that it was lower (p ≤ 0.05) for composite films with 3.5% (w/w of chitosan) zeolite. The brightness and b* value of the films decreased with the addition of zeolite at 3.5% (w/w of chitosan) for a same polymer concentration. The opacity did not change significantly (p > 0.05), although there was a tendency to decrease with the increase in the chitosan concentration.