Effects of Cymatocarpus solearis (Trematoda: Brachycoeliidae) on its second intermediate host, the Caribbean spiny lobster Panulirus argus.

Many digenean trematodes require three hosts to complete their life cycle. For Cymatocarpus solearis (Brachycoeliidae), the first intermediate host is unknown; the Caribbean spiny lobster Panulirus argus is a second intermediate host, and the loggerhead turtle Caretta caretta, a lobster predator, is the definitive host. Trophically-transmitted parasites may alter the behavior or general condition of intermediate hosts in ways that increase the hosts' rates of consumption by definitive hosts. Here, we examined the effects of infection by C. solearis on P. argus by comparing several physiological and behavioral variables among uninfected lobsters (0 cysts) and lobsters with light (1-10 cysts), moderate (11-30 cysts), and heavy (>30 cysts) infections. Physiological variables were hepatosomatic index, growth rate, hemocyte count, concentration in hemolymph of cholesterol, protein, albumin, glucose, dopamine (DA) and serotonin (5-HT). Behavioral variables included seven components of the escape response (delay to escape, duration of swimming bout, distance traveled in a swimming bout, swim velocity, acceleration, force exerted, and work performed while swimming). There was no relationship between lobster size or sex and number of cysts. Significant differences among the four lobster groups occurred only in concentration of glucose (lower in heavily infected lobsters) and 5-HT (higher in heavily and moderately infected lobsters) in plasma. As changes in 5-HT concentration can modify the host's activity patterns or choice of microhabitat, our results suggest that infection with C. solearis may alter the behavior of spiny lobsters, potentially increasing the likelihood of trophic transmission of the parasite to the definitive host.

Immune Response to Natural and Experimental Infection of Panulirus argus Virus 1 (PaV1) in Juveniles of Caribbean Spiny Lobster.

Experimental infections have been used to better comprehend the immune system of organisms, and to probe for additives that generate greater resistance and help reduce antibiotic use in aquaculture. We compared the immune response of juveniles of the Caribbean spiny lobster, Panulirus argus, infected naturally with Panulirus argus virus 1 (PaV1) versus organisms infected experimentally, to determine the analogy between both infectious processes. The immunological response was measured by hemagglutination activity, hemocyte count, and total phenoloxidase activity in plasma and hemocytes in 211 individuals that were either naturally infected (110), or had been injected with viral inoculum and followed for six months (101). The samples were classified into the following four groups according to the severity of the infection: 0, uninfected; 1, lightly; 2, moderately; and 3, severely infected), which was determined on the basis of PCR and histological criteria. A permutational MANOVA showed that both the origin (natural and experimental), and the severity of the infection contributed significantly to explain the variation in the immune response of lobsters. The lack of significance of the interaction term indicated that the immunological response changed with the severity of the infection in a similar way, regardless of its origin. The results of the present study suggest that the experimental viral infection of PaV1 produces a defense response similar to the natural pathways of contagion, and provides the bases to validate an immunological challenge protocol for the first time in crustaceans. The discussion includes the perspective of the conceptual models of immune response within an ecological context.

Alterations in the gut-associated microbiota of juvenile Caribbean spiny lobsters Panulirus argus (Latreille, 1804) infected with PaV1.

The spiny lobster Panulirus argus (Latreille, 1804) is currently affected by an unenveloped, icosahedral, DNA virus termed Panulirus argus virus 1 (PaV1), a virulent and pathogenic virus that produces a long-lasting infection that alters the physiology and behaviour of heavily infected lobsters. Gut-associated microbiota is crucial for lobster homeostasis and well-being, but pathogens could change microbiota composition affecting its function. In PaV1 infection, the changes of gut-associated microbiota are yet to be elucidated. In the present study, we used high-throughput 16S rRNA sequencing technology to compare the bacterial microbiota in intestines of healthy and heavily PaV1-infected male and female juveniles of spiny lobsters P. argus captured in Puerto Morelos Reef lagoon, Quintana Roo, Mexico. We found that basal gut-associated microbiota composition showed a sex-dependent bias, with females being enriched in amplicon sequence variants (ASVs) assigned to Sphingomonas, while males were enriched in the genus Candidatus Hepatoplasma and Aliiroseovarius genera. Moreover, the alpha diversity of microbiota decreased in PaV1-infected lobsters. A significant increase of the genus Candidatus Bacilloplasma was observed in infected lobsters, as well as a significant decrease in Nesterenkonia, Caldalkalibacillus, Pseudomonas, Cetobacterium and Phyllobacterium. We also observed an alteration in the abundances of Vibrio species. Results from this study suggest that PaV1 infection impacts intestinal microbiota composition in Panulirus argus in a sex-dependent manner.

iTRAQ-Based Proteomic Profile Analysis of the Hepatopancreas of Caribbean Spiny Lobsters Infected With Panulirus argus Virus 1: Metabolic and Physiological Implications.

The Caribbean spiny lobster Panulirus argus (Latreille, 1084) sustains economically valuable fisheries throughout the wider Caribbean region. This species is currently affected by the pathogenic virus Panulirus argus Virus 1 (PaV1) that causes a systemic and chronic-degenerative infection in juvenile spiny lobsters P. argus. To date, there is no available information regarding the host alterations induced by this pathogen at the molecular level. In the present study, comparative proteomic analyses of the changes in the hepatopancreas between infected and non-infected juvenile lobsters were analyzed by isobaric tags for relative and absolute quantitation (iTRAQ) coupled to synchronous precursor selection (SPS)-based MS3. We identified a total of 636 proteins, being 68 down-regulated and 71 up-regulated proteins. Among the down-regulated proteins, we identified several enzymes involved in the metabolism of hormones and lipids, digestive proteases and glycosidases, while proteins associated with the histone core, protein synthesis, immune response and RNA regulation were up-regulated. Several misregulated enzymes involved in the regulation of neuromodulators were also identified. RT-qPCR assays were used to validate the expression of transcripts encoding for selected differential proteins that were in concordance to proteomic data, as well as the tendency observed in the enzymatic activities of trypsin, chymotrypsin, and glycosidase. In a similar way, we observed glycogen reduction in muscle, and an increase in plasma acylglycerides and glucose, which may be explained by proteomic data. This study provides the first insight into the molecular changes in the hepatopancreas of Caribbean spiny lobsters associated to PaV1 infection. Data provided herein would help to clarify the origin of the molecular misregulations observed at macroscopic level in this host-pathogen interaction.

Do ecological characteristics drive the prevalence of Panulirus argus virus 1 (PaV1) in juvenile Caribbean spiny lobsters in a tropical reef lagoon?

PaV1 is a pathogenic virus found only to infect Caribbean spiny lobsters Panulirus argus, a major fishing resource. P. argus is a benthic mesopredator and has a complex life history, with several ontogenetic habitat changes. Habitat characteristics and species diversity of surrounding communities may have implications for disease dynamics. This is of more concern for juvenile lobsters, which are more susceptible to PaV1 and are far less mobile than adult lobsters. We targeted a population of juvenile P. argus in a reef lagoon in Mexico, where PaV1 was first observed in 2001. Prevalence has been since irregularly assessed, but in 2016 we began a more systematic assessment, with two sampling periods per year (June and November) in three different zones of the reef lagoon. To examine the relationship between PaV1 prevalence and potential ecological determinants, we assessed habitat complexity, cover of different substrates, and invertebrate community composition in all zones during the first four sampling periods (June and November 2016 and 2017). Habitat complexity and percent cover of some substrates varied with zone and sampling period. This was the case for seagrass and macroalgae, which nevertheless were the dominant substrates. The invertebrate community composition varied with sampling period, but not with zone. Probability of infection decreased with increasing lobster size, consistent with previous studies, but was not affected by zone (i.e., variations in ecological characteristics did not appear to be sufficiently large so as to influence prevalence of PaV1). This result possibly reflects the dominance of marine vegetation and suggests that lobsters can be sampled throughout the reef lagoon to assess PaV1 prevalence. Prevalence was higher in only one of seven sampling periods (November 2017), suggesting that the pathogen has leveled off to an enzootic level.

Confirming validity measures of visual assessment of PaV1 infection in Caribbean spiny lobsters.

Panulirus argus virus 1 (PaV1) affects wild populations of Caribbean spiny lobsters. PaV1 can be lethal but shows predilection for juvenile lobsters. Because P. argus is one of the most valuable fisheries around the wider Caribbean region, monitoring disease prevalence in local populations is desirable. Diseased lobsters are easily recognized by their milky hemolymph, but this sign only becomes evident in advanced stages of infection. Other methods have been developed to detect PaV1, but are less practical for long-term monitoring of patterns of infection in populations. A previous study estimated the validity measures (sensitivity and specificity) of detection of PaV1 infection by observed clinical signs against endpoint PCR assays, using a representative sample of lobsters comprising mainly subadults and adults from a commercial fishing area. In the present study, these validity measures were estimated in a similar manner for a different population comprising mainly juveniles from a protected nursery area. We obtained virtually the same sensitivity and specificity values (0.48 and 1, respectively) for observed clinical signs as in the previous study (0.51 and 1, respectively), confirming the validity of applying a simple 2× correction factor to monitor the patterns of PaV1 infection over time based on more easily conducted visual assessments of a representative sample of the population.

Morphometric relationships and seasonal variation in size, weight, and a condition index of post-settlement stages of the Caribbean spiny lobster.

Spiny lobsters have a protracted pelagic, oceanic larval phase. The final larval stage metamorphoses into a non-feeding postlarva (puerulus) that actively swims towards the coast to settle in shallow habitats and does not resume feeding until after the molt into the first-stage juvenile. Therefore, the body dimensions and nutritional condition of both settled pueruli and first juveniles are likely to vary over time, potentially playing a crucial role in the recruitment to the benthic population. We compared carapace length (CL), height (CH), and width (CW); total length (TL), and body weight (W) between pueruli and first juveniles of the Caribbean spiny lobster, Panulirus argus, as well as morphometric relationships between both developmental stages. Except for CL, all other dimensions were larger in first juveniles, but more markedly CH and W. The slopes of the CH vs CL, CW vs CL, and W vs CL regressions differed significantly between stages, and all log-transformed relationships showed isometry in both stages, except for the CH vs CL relationship, which showed positive allometry. These results reflect a morphological change from the flatter, more streamlined body of the puerulus, to the heavier, more cylindrical body of the juvenile. We also analyzed seasonal variations in CL, W, the W/CL index (a morphometric condition index), and a modified W/CL index (i.e. after controlling for a significant effect of CL) of both stages using individuals monthly collected over 12 consecutive seasons (Autumn 2010-Summer 2013). In both stages, all three variables exhibited significant seasonal variation. For pueruli, the modified W/CL index differed from average in only two seasons, winter 2011 (higher) and summer 2013 (lower), but showed great within-season variation (larger coefficients of variation, CV), potentially reflecting variability in nutritional condition of larvae prior to metamorphosis and in the distances swum by individual pueruli to the settlement habitats. For first juveniles, the modified W/CL index was higher than average in winter and spring 2011, and lower in autumn 2011 and winter 2012, but showed less within season variation (smaller CVs), suggesting a combination of carry-over effects of puerulus condition and effects of local conditions (e.g., food availability and predation risk). These findings warrant further investigation into factors potentially decoupling settlement from recruitment processes.

Diversity and abundance of conspicuous macrocrustaceans on coral reefs differing in level of degradation.

Coral reefs sustain abundant and diverse macrocrustaceans that perform multiple ecological roles, but coral reefs are undergoing massive degradation that may be driving changes in the species composition and abundance of reef-associated macrocrustaceans. To provide insight into this issue, we used non-destructive visual census techniques to compare the diversity and abundance of conspicuous macrocrustaceans (i.e., those >1 cm and visible without disturbance) between two shallow Caribbean coral reefs similar in size (∼1.5 km in length) and close to each other, but one ("Limones") characterized by extensive stands of the branching coral Acropora palmata, and the other ("Bonanza") dominated by macroalgae and relic coral skeletons and rubble (i.e., degraded). We also assessed the structural complexity of each reef and the percent cover of various benthic community components. Given the type of growth of A. palmata, we expected to find a greater structural complexity, a higher cover of live coral, and a lower cover of macroalgae on Limones, and hence a more diverse and abundant macrocrustacean community on this reef compared with Bonanza. Overall, we identified 63 macrocrustacean species (61 Decapoda and two Stomatopoda). Contrary to our expectations, structural complexity did not differ significantly between the back-reef zones of these reefs but varied more broadly on Limones, and the diversity and abundance of macrocrustaceans were higher on Bonanza than on Limones despite live coral cover being higher on Limones and macroalgal cover higher on Bonanza. However, the use of various types of microhabitats by macrocrustaceans differed substantially between reefs. On both reefs, the dominant species were the clinging crab Mithraculus coryphe and the hermit crab Calcinus tibicen, but the former was more abundant on Bonanza and the latter on Limones. M. coryphe occupied a diverse array of microhabitats but mostly coral rubble and relic skeletons, whereas C. tibicen was often, but not always, found associated with colonies of Millepora spp. A small commensal crab of A. palmata, Domecia acanthophora, was far more abundant on Limones, emerging as the main discriminant species between reefs. Our results suggest that local diversity and abundance of reef-associated macrocrustaceans are partially modulated by habitat degradation, the diversity of microhabitat types, and the establishment of different commensal associations rather than by structural complexity alone.

Habitat degradation alters trophic pathways but not food chain length on shallow Caribbean coral reefs.

Habitat degradation can affect trophic ecology by differentially affecting specialist and generalist species, and the number and type of interspecific relationships. However, the effects of habitat degradation on the trophic ecology of coral reefs have received limited attention. We compared the trophic structure and food chain length between two shallow Caribbean coral reefs similar in size and close to each other: one dominated by live coral and the other by macroalgae (i.e., degraded). We subjected samples of basal carbon sources (particulate organic matter and algae) and the same 48 species of consumers (invertebrates and fishes) from both reefs to stable isotope analyses, and determined the trophic position of consumers and relative importance of various carbon sources for herbivores, omnivores, and carnivores. We found that both reefs had similar food chain length and trophic structure, but different trophic pathways. On the coral-dominated reef, turf algae and epiphytes were the most important carbon source for all consumer categories, whereas on the degraded reef, particulate organic matter was a major carbon source for carnivores. Our results suggest that the trophic structure of the communities associated with these reefs is robust enough to adjust to conditions of degradation.

Variability in prevalence of Cymatocarpus solearis (Trematoda, Brachycoeliidae) in Caribbean spiny lobsters Panulirus argus (Decapoda: Palinuridae) from Bahía de la Ascensión (Mexico).

Cymatocarpus solearis, a brachycoeliid trematode that parasitizes sea turtles, uses spiny lobsters Panulirus argus as second intermediate hosts. We examined variability in infection by C. solearis in Bahía de la Ascensión, Mexico, a tropical bay with a highly productive fishery for P. argus that is based on the extensive use of casitas (large artificial shelters), which can harbor multiple juveniles and adults of this gregarious lobster species. We sampled 3956 lobsters from 466 casitas distributed over three bay zones within two closed seasons and one fishing season. In these lobsters (9.5-115.2mm carapace length, CL), the average infection prevalence was 11.7% but the probability of infection increased significantly with size. Prevalence varied with season but was consistently higher in one zone than in the other two zones. Infection with C. solearis was negatively related with clinical infection with Panulirus argus Virus 1 (PaV1). We also sampled 405 lobsters from the commercial catch obtained throughout the bay at the onset of the fishing season. In these relatively larger lobsters (70.3-168.0mm CL), average prevalence of C. solearis was higher (23.5%) but was not affected by lobster size or sex. Encysted metacercariae occurred in both abdominal and cephalothoracic muscles. The effects of C. solearis on phenotypic traits of P. argus that may potentially impact the host population dynamics and fisheries remain to be investigated.

Panulirus argus virus 1 (PaV1) infection prevalence and risk factors in a Mexican lobster fishery employing casitas.

In Bahía de la Ascensión in Mexico, the Caribbean spiny lobster Panulirus argus fishery is based on extensive use of artificial shelters (casitas) that can harbor both juveniles and adults of this highly gregarious species. There is concern that the use of casitas might increase contact transmission of Panulirus argus virus 1 (PaV1). However, a previous study found no evidence for lobster crowding within casitas influencing the prevalence of clinical PaV1 disease, although differences in clinical prevalence were noted between different bay environments. To investigate this more closely, 683 lobsters were sampled from casitas in 2 zones in this bay (Vigía Chico, a shallow low-vegetation zone, and Punta Allen, a deeper dense-vegetation zone) previously found to have the lowest and highest prevalence, respectively, of observed clinical signs. When hemolymph collected from these lobsters was tested by PCR, the prevalence of PaV1 infection was found to be significantly lower in Vigía Chico relative to Punta Allen irrespective of season or the size, sex, or presence of shell injuries on lobsters. Among 714 large commercial-catch lobsters collected throughout the bay, the prevalence of infection was low irrespective of year or sex. For all lobsters tested, the sensitivity (0.510) at which PaV1 infection was detected by observed clinical signs was about half that determined by PCR, but the specificity of clinical signs was absolute (1), indicating that a simple 2× correction factor can be used to accurately estimate PaV1 infection prevalence based on more easily conducted visual assessments of lobsters.

Physiological and immunological characterization of Caribbean spiny lobsters Panulirus argus naturally infected with Panulirus argus Virus 1 (PaV1).

The present study compares 13 physiological and immunological variables between a group of healthy Panulirus argus lobsters and a group of lobsters naturally infected with Panulirus argus Virus 1 (PaV1). Viral infection was determined through histopathology and PCR. Ten of the 13 variables differed significantly between the 2 groups. Using these variables, a principal component analysis yielded 2 separate clusters: one corresponding to the healthy group and the other corresponding to the infected group. In particular, infected lobsters exhibited significantly lower levels of osmotic pressure, total hemocyte counts, plasmatic proteins, and total phenoloxidase (PO) activity in plasma, as well as significantly higher levels of cholesterol and acylglycerides. These features are consistent with metabolic wasting, hyperlipidemia, and presumed immune suppression. Infection with PaV1 appears to increase the susceptibility of lobsters to some other opportunistic pathogens, as 61.1% of infected lobsters presented infestations of ciliate epibionts (Epystilis and Zoothamniun) in the gill chamber compared with 11.5% lobsters in the healthy group. Infected lobsters also showed significantly higher levels of total PO activity in degranulated hemocytes and trypsin inhibitor activity, potentially indicating activation of immune response by the PO system during the systemic infection with PaV1.

Variability in clinical prevalence of PaV1 in Caribbean spiny lobsters occupying commercial casitas over a large bay in Mexico.

In Bahía de la Ascensión, Mexico, the fishery for spiny lobsters Panulirus argus is based on the extensive use of casitas, large artificial shelters that can harbor the full size range of these highly gregarious lobsters. The discovery of a pathogenic virus in these lobsters (Panulirus argus virus 1, or PaV1) has raised concern about its potential effects on casita-based fisheries. Because in Bahía de la Ascensión visibly infected lobsters represent an immediate loss of revenue, we examined variability in clinical prevalence of PaV1 (percentage of lobsters visibly infected) in thousands of lobsters sampled from the commercial catch at the onset of 3 consecutive fishing years, and from 530 casitas distributed over 3 zones within the bay during 2 fishing and 2 closed seasons. In the commercial catch (lobsters 67 to 147 mm carapace length [CL]), clinical prevalence of PaV1 was low and was not affected by year or sex. In lobsters (9.2 to 115.0 mm CL) that occupied casitas, clinical prevalence of PaV1 varied with sampling season and was always higher in juveniles than in subadults or adults, but was consistently lower in one zone relative to the other 2 zones. The average clinical prevalence of PaV1 in this bay was statistically similar to the average clinical prevalence reported in Cuba, where casitas are also used, and in Florida Bay, USA, where casitas are not used. To date, PaV1 has had no discernible impact on the lobster fishery in Bahía de la Ascensión, suggesting that clinical prevalence is not influenced by the use of casitas per se.

Influence of local habitat features on disease avoidance by Caribbean spiny lobsters in a casita-enhanced bay.

In Bahía de la Ascensión, Mexico, 'casitas' (large artificial shelters) are extensively used to harvest Caribbean spiny lobsters Panulirus argus. After the discovery of a pathogenic virus, Panulirus argus virus 1 (PaV1), in these lobsters, laboratory experiments revealed that PaV1 could be transmitted by contact and through water, and that lobsters avoided shelters harboring diseased conspecifics. To examine these issues in the context of casitas, which typically harbor multiple lobsters of all sizes, we examined the distribution and aggregation patterns of lobsters in the absence/presence of diseased conspecifics (i.e. visibly infected with PaV1) in 531 casitas distributed over 3 bay zones, 1 poorly vegetated ('Vigía Chico', average depth: 1.5 m) and 2 more extensively vegetated ('Punta Allen': 2.5 m; 'Los Cayos': 2.4 m). All zones had relatively high indices of predation risk. Using several statistical approaches, we found that distribution parameters of lobsters were generally not affected by the presence of diseased conspecifics in casitas. Interestingly, however, in the shallower and less vegetated zone (Vigía Chico), individual casitas harbored more lobsters and lobsters were actually more crowded in casitas containing diseased conspecifics, yet disease prevalence was the lowest in lobsters of all sizes. These results suggest that (1) investment in disease avoidance by lobsters is partially modulated by local habitat features, (2) contact transmission rates of PaV1 may be lower in nature than in the laboratory, and (3) water-borne transmission rates may be lower in shallow, poorly vegetated habitats more exposed to solar ultraviolet radiation, which can damage viral particles.

Detection of Panulirus argus Virus 1 (PaV1) in exported frozen tails of subadult-adult Caribbean spiny lobsters Panulirus argus.

The Caribbean spiny lobster Panulirus argus is a valuable fishing resource and the trade in frozen lobster tails is an important industry. However, the presence of the pathogenic virus Panulirus argus Virus 1 (PaV1), which causes systemic infection in P. argus and is particularly lethal to juvenile individuals, has not been previously examined in imported/exported lobster products. We used PCR assays to determine the presence of PaV1 in abdominal muscle tissue of 22 frozen P. argus tails exported from Belize to Mexico. Based on their size, the tails belonged to subadult-adult lobsters. Using specific primers targeted for PaV1 resulted in 11 tails showing a specific 499 bp band. The sequence of positive amplified fragments showed a high similarity to PaV1 (95% identity with GenBank accession no. EF206313.1). Although the pathogenicity of PaV1 was not evaluated in the present study, our results provide the first evidence of PaV1 in frozen lobster tails exported in the seafood industry as well as the first molecular evidence of PaV1 in adult lobsters.

Influence of conspecific and heterospecific aggregation cues and alarm odors on shelter choice by syntopic spiny lobsters.

In spiny lobsters, conspecific scents ("aggregation cues") may mediate gregarious diurnal sheltering, but scents from injured conspecifics ("alarm odors") may elicit avoidance behavior. In laboratory experiments, individuals of two coexisting species, Panulirus guttatus (a reef-obligate) and P. argus (a temporary reef-dweller), significantly chose shelters emanating conspecific aggregation cues and responded randomly to shelters emanating heterospecific aggregation cues. However, despite evidence that the two species perceived each other's alarm odors to a similar extent, P. guttatus responded randomly to shelters emanating either conspecific or heterospecific alarm odors, whereas P. argus significantly avoided both. This differential influence of alarm odors likely reflects interspecific differences in life history, sociality, and behavior. The less social, reef-obligate P. guttatus lobsters forage close to their reef dens, into which they retract deeply upon perception of risk. This cryptic behavior may offset the need to avoid conspecific (and heterospecific) alarm odors. In contrast, avoidance of conspecific alarm odors by P. argus is consistent with its ontogenetic habitat shifts and greater sociality. Furthermore, because reef-dwelling P. argus lobsters forage across open areas away from the reef, an ability to avoid alarm odors from P. guttatus upon returning to their reef dens may increase their fitness.

Prevalence of Panulirus argus Virus 1 (PaV1) and habitation patterns of healthy and diseased Caribbean spiny lobsters in shelter-limited habitats.

Caribbean spiny lobsters Panulirus argus are socially gregarious, preferring shelters harboring conspecifics over empty shelters. In laboratory trials, however, healthy lobsters strongly avoided shelters harboring lobsters infected with the highly pathogenic Panulirus argus Virus 1 (PaV1). Because PaV1 is transmitted by contact, this behavior may thwart its spread in wild lobsters. In a field experiment conducted from 1998 to 2002 in a shelter-poor reef lagoon (Puerto Morelos, Mexico), densities of juvenile P. argus increased significantly on sites enhanced with artificial shelters (casitas) but not on control sites. Because PaV1 emerged in this location during 2000, we reexamined these data to assess whether casitas could potentially increase transmission of PaV1. In 2001, PaV1 prevalence was 2.5% and the cohabitation level (percentage of healthy lobsters cohabiting with diseased lobsters) was similar between natural shelters (3.5%) and casitas (2.4 %). The relative lobster densities in casita sites and control sites did not change significantly before (1998-1999) or after (2001-2002) the disease emergence. In late 2006, data from casita sites showed a significant increase in prevalence (10.9%) and cohabitation level (29.4%), but no significant changes in lobster density. In May 2006, casitas were deployed on shelter-poor sites within Chinchorro Bank, 260 km south of Puerto Morelos. In late 2006, prevalence and cohabitation level were 7.4 and 21.7%, respectively. Our results are inconclusive as to whether or not casitas increase PaV1 transmission, but suggest that across shelter-poor habitats, lobsters make a trade-off between avoiding diseased conspecifics and avoiding predation risk.

Presence of Panulirus argus virus 1 (PaV1) in juvenile spiny lobsters Panulirus argus from the Caribbean coast of Mexico.

Macroscopic evidence, histological sections, transmission electron microscopy (TEM) evaluation, and PCR analyses of 25 apparently diseased juvenile spiny lobsters Panulirus argus from the reef lagoon of Puerto Morelos, Mexico, showed the presence of Panulirus argus Virus 1 (PaV1). Cowdry Type A intranuclear viral inclusions were observed in histological analyses, icosahedral viral particles were observed by TEM, and PCR using specific primers for PaV1 amplified a fragment of 499 bp. This is the first report of PaV1 infecting P. argus outside the Florida Keys, USA.

Enhancement of juvenile Caribbean spiny lobsters: an evaluation of changes in multiple response variables with the addition of large artificial shelters.

Shortage of natural crevice shelters may produce population bottlenecks in juvenile Caribbean spiny lobsters (Panulirus argus), a socially gregarious species. We conducted a field experiment to test enhancement of a local population of juvenile P. argus with the addition of artificial shelters ("casitas") that mimic large crevices (1.1 m(2) in surface area and 3.8 cm in height). Changes in density and biomass of juvenile lobsters 15-50 mm carapace length (CL) were assessed with a multiple before-after control-impact (MBACI) analysis. Separate analyses were also conducted on small (15-35 mm CL) and large (35.1-50 mm CL) juveniles to assess size-related effects. First, we carried out 13 lobster surveys on nine fixed 1-ha sites over a shallow reef lagoon ("before" period). Then, we deployed ten casitas in each of five sites and left four sites as controls, and conducted 22 further surveys ("after" period). Deployment of casitas resulted in a sixfold increase in juvenile density (76% contributed by small and 24% by large juveniles) and a sevenfold increase in biomass (40 and 60%, respectively). Capture-recapture results revealed that enhancement was achieved not by promoting individual growth but by increasing survival, persistence, and foraging ranges of small and large juveniles. Casitas both mitigated shortage of natural shelter and increased sociality, allowing for cohabitation of smaller, more vulnerable juveniles with larger conspecifics that have greater defensive abilities. Casitas may help enhance local populations of juvenile P. argus in Caribbean seagrass habitats, typically poor in natural crevice shelters. The use of MBACI and the simultaneous assessment of multiple interrelated response variables may be a powerful analytical approach to test shelter limitation in other species and to examine the function of structural habitat in other systems.

Factors affecting growth of the spiny lobsters Panulirus gracilis and Panulirus inflatus (Decapoda: Palinuridae) in Guerrero, México

The effects of sex, injuries, season and site on the growth of the spiny lobsters Panulirus gracilis, and P. inflatus, were studied through mark-recapture techniques in two sites with different ecological characteristics on the coast of Guerrero, México. Panulirus gracilis occurred in both sites, whereas P. inflatus occurred only in one site. All recaptured individuals were adults. Both species had similar intermolt periods, but P. gracilis had significantly higher growth rates (mm carapace length week-1) than P. inflatus as a result of a larger molt increment. Growth rates of males were higher than those of females in both species owing to larger molt increments and shorter intermolt periods in males. Injuries had no effect on growth rates in either species. Individuals of P. gracilis grew faster in site 1 than in site 2. Therefore, the effect of season on growth of P. gracilis was analyzed separately in each site. In site 2, growth rates of P. gracilis were similar in summer and in winter, whereas in site 1 both species had higher growth rates in winter than in summer. This could be due to spatial differences in processes related to changes in population density and food resources, which were documented in previous works. The overall results show that P. gracilis grows faster than P. inflatus, and that growth rates of both species are highly variable and are affected by environmental factors such as site and season, which should be taken into account when attempting to produce population growth curves for each species.