Behavioral Immunity Suppresses an Epizootic in Caribbean Spiny Lobsters

Sociality has evolved in a wide range of animal taxa but infectious diseases spread rapidly in populations of aggregated individuals, potentially negating the advantages of their social interactions. To disengage from the coevolutionary struggle with pathogens, some hosts have evolved various forms of “behavioral immunity”; yet, the effectiveness of such behaviors in controlling epizootics in the wild is untested. Here we show how one form of behavioral immunity (i.e., the aversion of diseased conspecifics) practiced by Caribbean spiny lobsters (Panulirus argus) when subject to the socially transmitted PaV1 virus, appears to have prevented an epizootic over a large seascape. We capitalized on a "natural experiment" in which a die-off of sponges in the Florida Keys (USA) resulted in a loss of shelters for juvenile lobsters over a ~2500km² region. Lobsters were thus concentrated in the few remaining shelters, presumably increasing their exposure to the contagious virus. Despite this spatial reorganization of the population, viral prevalence in lobsters remained unchanged after the sponge die-off and for years thereafter. A field experiment in which we introduced either a healthy or PaV1-infected lobster into lobster aggregations in natural dens confirmed that spiny lobsters practice behavioral immunity. Healthy lobsters vacated dens occupied by PaV1-infected lobsters despite the scarcity of alternative shelters and the higher risk of predation they faced when searching for a new den. Simulations from a spatially-explicit, individual-based model confirmed our empirical results, demonstrating the efficacy of behavioral immunity in preventing epizootics in this system.     

Diseases of Nephrops and Metanephrops: a review

Nephrops and Metanephrops are commercially exploited genera within the family Nephropidae (clawed lobsters). Commercial fisheries for each genus exist in the Northern and Southern Hemispheres and utilise trawling or trapping for capture. Despite a relative lack of dedicated disease surveys on lobsters from these fisheries, several important symbionts and pathogens have been described. The most significant known pathogen of Metanephrops (challengeri) is a microsporidian parasite (Myospora metanephrops) which causes destruction of the skeletal and heart muscles of infected lobsters while the most significant known pathogen of Nephrops (norvegicus) is a dinoflagellate parasite assigned to the genus Hematodinium. This parasite has been responsible for an ongoing epidemic in fished populations of N. norvegicus in Northern Europe since at least the early 1980s and since then extensive studies on its life history and pathogenesis have occurred. Despite these research efforts significant gaps exist in our knowledge of the effects of parasites such as Hematodinium on the fished and non-fished portions of Nephrops populations and on the effect of fishery practices on the spread of infection. Furthermore, little is known about the effect of this (and other) pathogens on cohort survivability and the likelihood that early life stages will be effectively recruited to the fishery. This review summarises the available literature on diseases of these two lobster genera and provides an assessment of future research needs in this discipline.     

Subtidal-intertidal trophic links: American lobsters [Homarus americanus (Milne-Edwards)] forage in the intertidal zone on nocturnal high tides

Homarus americanus (Milne-Edwards), the American lobster, is a predator in New England subtidal communities, feeding on ecologically important grazers (sea urchins), mesopredators (crabs), and basal species (mussels). In this study, we provide the first report of adult American lobsters foraging in rocky intertidal habitats during nocturnal high tides. Censuses by SCUBA divers in the low intertidal (Chondrus crispus Stackhouse) zone showed mean densities of 2.2 lobsters/20 m² on nocturnal high tides, with contrasting low densities of 0.18/20 m² during diurnal high tides. Nocturnal high-tide intertidal densities were 62% of those reported in a previous study of lobsters in nearby subtidal rocky areas (Novak, 2004). The average carapace length of lobsters in the intertidal at night was > 50 mm. These lobsters were actively foraging in the intertidal with collected individuals having a mean stomach fullness of 67%. Prey found in the stomach contents primarily consisted of crabs, mussels and snails. Field experiments showed that lobsters rarely fed on medium to large size individuals of the common intertidal snail, Littorina littorea (L.). In contrast, experiments with local crab species demonstrated that lobsters actively and readily prey on Cancer irroratus (Say) and Carcinus maenas (L.), but were significantly less likely to consume Cancer borealis (Stimpson). The abundance of Carcinus maenas and blue mussels (Mytilus edulis L.) in the intertidal zone may explain the upshore movement of lobsters. Since nocturnal migration of Homarus americanus into the intertidal zone has not been documented before, our understanding of the dynamics of New England intertidal communities needs to be expanded to include this predator.     

Agonistic interactions between invasive green crabs, Carcinus maenas (Linnaeus), and sub-adult American lobsters, Homarus americanus (Milne Edwards)

The invasive green crab, Carcinus maenas, has recently expanded its range into the Southern Gulf of St. Lawrence, where there is potential for substantial niche overlap with juvenile American lobsters, Homarus americanus. We used two experiments to elicit, record and analyze the agonistic interactions of adult green crabs (carapace width of 63-75 mm) and sub-adult (carapace length of 55-70 mm) lobsters. The first experiment gave each animal equal access to a limited food resource. The green crabs were first to the food in significantly more trials, spent a significantly greater proportion of time with the food, and were able to successfully defend the food from attacks by the heavier lobsters. In the second experiment, we allowed the lobsters to gain possession and initiate feeding on the food before releasing the green crabs. In these trials, the lobsters spent significantly more time with the food, and were able to defend the food from the green crabs. The results of both experiments are discussed in the context of game theory. The different behaviour of the crustaceans in the two experiments is consistent with the “bourgeois” strategy in a hawk and dove game simulation. With this strategy, an animal acts like a hawk if in possession of a resource, but acts like a dove if the other animal is in possession of the resource. The fact that the green crabs were able to physically compete with, and in many cases dominate the larger, heavier lobsters supports the potential for competitive impacts of green crabs on sub-adult lobsters.     

Autophagy plays an important role in protecting Pacific oysters from OsHV-1 and Vibrio aestuarianus infections

Recent mass mortality outbreaks around the world in Pacific oysters, Crassostrea gigas, have seriously affected the aquaculture economy. Although the causes for these mortality outbreaks appear complex, infectious agents are involved. Two pathogens are associated with mass mortality outbreaks, the virus ostreid herpesvirus 1 (OsHV-1) and the bacterium Vibrio aestuarianus. Here we describe the interactions between these 2 pathogens and autophagy, a conserved intracellular pathway playing a key role in innate immunity. We show for the first time that autophagy pathway is present and functional in Pacific oysters and plays an important role to protect animals from infections. This study contributes to better understand the innate immune system of Pacific oysters.     

Parasitic diseases in marine cage culture--an example of experimental evolution of parasites?

Rapid development of fish culture in marine cages has been associated with an emergence of parasitic diseases. There is a general trend to an increase in infections with ectoparasites with direct life cycles and a reduced diversity of parasites in aquaculture. Some mariculture creates conditions that are similar to serial passage experiments, which are used to study adaptation during experimental evolution of pathogens. In particular, increased density of fish, repeated introduction of naive hosts, homogenous host populations, fast growth and a potential decrease in genetic diversity are attributes of both aquaculture and serial passage experiments. Some free-living organisms, for example Neoparamoeba spp. and Uronema spp. parasitise fish in culture, but have not been reported from wild populations. Farming fish in marine cages can increase the risk of outbreaks of parasitic diseases, including those caused by opportunistic parasites. However, aquaculture has the potential to control parasitic diseases through selective breeding, vaccination and general fish health management.     

The hypervariable domain of the mitochondrial control region in Atlantic spiny lobsters and its potential as a marker for investigating phylogeographic structuring

Atlantic spiny lobsters support major fisheries in northeastern Brazilian waters and in the Caribbean Sea. To avoid reduction in diversity and elimination of distinct stocks, understanding their population dynamics, including structuring of populations and genetic diversity, is critical. We here explore the potential of using the hypervariable domain in the control region of the mitochondrial DNA as a genetic marker to characterize population subdivision in spiny lobsters, using Panulirus argus as the species model. The primers designed on the neighboring conserved genes have amplified the entire control region (approx. 780 bases) of P. argus and other closely related species. Average nucleotide and haplotype diversity within P. argus were found to be high, and population structuring was hypothesized. The data suggest a division of P. argus into genetically different phylogeographic groups. The hypervariable domain seems to be useful for determining genetic differentiation of geographically distinct stocks of P. argus and other Atlantic spiny lobsters.     

Identification of the pathogens associated with skin ulceration and peristome tumescence in cultured sea cucumbers Apostichopus japonicus (Selenka)

The aquaculture of sea cucumbers Apostichopus japonicus (Selenka) has developed rapidly in China in recent years, but is increasingly affected by diseases such as skin ulceration and peristome tumescence. Previous studies on the pathogens causing these diseases focused largely on bacterial causes. In December 2008, we isolated four dominant bacterial species from lesions present in A. japonicus with the aforementioned diseases, from a farm in Yangkou (Qingdao, China). With two of these bacterial species, experimental infection of healthy A. japonicus resulted in the same disease symptoms that occurred in naturally infected A. japonicus. These two species were identified as Pseudoalteromonas sp. and Pseudoalteromonas tetraodonis. The early symptoms of infection for these bacterial species were ulcer spots on the dorsal skin and abdominal parapodia, followed by an increase in the number of ulcer spots or their merging into larger spots. Additionally, we isolated a spherical virus 100-250 nm in diameter and with a bilayer capsule, from A. japonicus with another disease from four different farms. By experimental infection with crude extracts of the virus, healthy laboratory-acclimatized A. japonicus developed the same symptoms as in natural infected cases. The early symptoms of viral infection comprised a decrease in tentacle activity, decay of dorsal papillate podia, peristome tumescence and abdominal ulceration. Our study demonstrates that the bacteria and virus were both responsible for skin ulceration and peristome tumescence in A. japonicus, but resulted in different early disease symptoms.     

Survey of bumble bee (Bombus) pathogens and parasites in Illinois and selected areas of northern California and southern Oregon

Pathogens have been implicated as potential factors in the recent decline of some North American bumble bee (Bombus) species, but little information has been reported about the natural enemy complex of bumble bees in the United States. We targeted bumble bee populations in a state-wide survey in Illinois and several sites in California and Oregon where declines have been reported to determine presence and prevalence of natural enemies. Based on our observations, most parasites and pathogens appear to be widespread generalists among bumble bee species, but susceptibility to some natural enemies appeared to vary.     

Agonistic interactions between the invasive green crab, Carcinus maenas (Linnaeus) and juvenile American lobster, Homarus americanus (Milne Edwards)

Invasive organisms have the potential for competition with native organisms. In the Southern Gulf of St. Lawrence, juvenile American lobsters have a potential spatial overlap with adult green crabs. Crustaceans use agonistic behaviour to settle disputes, with the larger organism often winning contests for limited resources such as food and shelter. Two experiments were carried out using adult green crabs (53-76 mm carapace width) and juvenile American lobsters (28-57 mm carapace length). The first experiment used a limited food resource. We found that green crabs were the first to the food in all trials, fed in significantly more trials than lobsters and spent a significantly greater proportion of time with the food. The lobsters were only able to displace the green crabs from the food in 2 of 65 attempts. The second experiment was designed to examine shelter competition; unexpectedly some predation by green crabs on lobsters occurred, which allowed us to test hypotheses about how relative size and shelter use affect predation. Green crabs captured and consumed juvenile lobsters in 6 of 11 trials. The lobsters that survived spent significantly more time in shelter. There was no clear relationship between shelter use and size of lobster. The lobsters that were larger in relation to the green crabs suffered a higher rate of predation, which we believe was due to more conspicuous activity and less use of shelter. It appears that green crabs have the potential to negatively impact native juvenile lobster.     

Review of Panulirus argus virus 1--a decade after its discovery

In 2000, a pathogenic virus was discovered in juvenile Caribbean spiny lobsters Panulirus argus from the Florida Keys, U.S.A. Panulirus argus virus 1 (PaV1) is the first naturally occurring pathogenic virus reported from lobsters, and it profoundly affects their ecology and physiology. PaV1 is widespread in the Caribbean with infections reported in Florida (U.S.A.), St. Croix, St. Kitts, Yucatan (Mexico), Belize, and Cuba. It is most prevalent and nearly always lethal in the smallest juvenile lobsters, but this declines with increasing lobster size; adults harbor the virus, but do not present the characteristic signs of the disease. No other PaV1 hosts are known. The prevalence of PaV1 in juvenile lobsters from the Florida Keys has been stable since 1999, but has risen to nearly 11% in the eastern Yucatan since 2001. Heavily infected lobsters become sedentary, cease feeding, and die of metabolic exhaustion. Experimental routes of viral transmission include ingestion, contact, and for newly settled juveniles, free virus particles in seawater. Prior to infectiousness, healthy lobsters tend to avoid diseased lobsters and so infected juvenile lobsters mostly dwell alone, which appears to reduce disease transmission. However, avoidance of diseased individuals may result in increased shelter competition between healthy and diseased lobsters, and greater predation on infected lobsters. Little is known about PaV1 outside of Mexico and the USA, but it poses a potential threat to P. argus fisheries throughout the Caribbean.     

Cryptosporidium and Giardia: Treatment options and prospects for new drugs

Cryptosporidium species and Giardia intestinalis are the most common enteric protozoan pathogens affecting humans worldwide. In recent years, nitazoxanide has been licensed in the United States for the treatment of cryptosporidiosis in non-immunodeficient children and adults, becoming the first drug approved for treating this disease. There is a need for a highly effective treatment for cryptosporidiosis in immunodeficient patients, but the quest for such a drug has proven to be elusive. While not effective against Cryptosporidium, nitroimidazoles such as metronidazole or tinidazole are effective treatments for giardiasis and can be administered as a single dose. Albendazole and nitazoxanide are effective against giardiasis but require multiple doses. Nitazoxanide is the first new drug developed for treating giardiasis in more than 20 years. New potentially promising drug targets in Cryptosporidium and Giardia have been identified, but there appears to be little activity toward clinical development of new drugs.     

内生真菌感染对宿主羊草抗病性的影响

以感染内生真菌的天然禾草羊草为实验材料,通过体外纯培养条件下的内生真菌、感染内生真菌的离体叶片和在体叶片对3种病原菌的抑菌实验,以探讨内生真菌对宿主植物羊草在抗病性方面的贡献。结果表明:体外纯培养条件下,分离自羊草的内生真菌Epichlobromicola对新月弯孢(Curvularia lunata)、根腐离蠕孢(Bipolaris sorokiniana)和枝孢霉(Cladosporium sp.)这3种病原菌都具有抑制作用,抑菌率分别达56.22%,46.93%和45.15%,且内生真菌培养滤液可以有效抑制这3种病原菌的孢子萌发,平均萌发率分别为30.4%,15.7%和16.4%;宿主植物叶片在离体条件下,内生真菌感染可以有效降低羊草叶片受C.lunata和C.sp.侵染后的病斑数或病斑长度,但对B.sorokiniana不起作用,甚至提高了叶片的病斑数及病斑长度,而离体叶片提取液对不同病原菌均有不同程度的抑制作用;在体条件下,内生真菌均可以通过降低叶片病斑数来增强羊草植株对这3种病原菌的抗性。由此看来,内生真菌E.bromicola对宿主植物羊草在抗病原菌侵染方面有一定的增益作用。     

The complex interactions between host immunity and non-biotrophic fungal pathogens of wheat leaves

Significant progress has been made in elucidating the mechanisms used by plants to recognize pathogens and activate “immune” responses. A “first line” of defense can be triggered through recognition of conserved Pathogen or Microbe Associated Molecular Patterns (PAMPs or MAMPs), resulting in activation of basal (or non-host) plant defenses, referred to as PAMP-triggered immunity (PTI). Disease resistance responses can also subsequently be triggered via gene-for-gene type interactions between pathogen avirulence effector genes and plant disease resistance genes (Avr-R), giving rise to effector triggered immunity (ETI). The majority of the conceptual advances in understanding these systems have been made using model systems, such as Arabidopsis, tobacco, or tomato in combination with biotrophic pathogens that colonize living plant tissues. In contrast, how these disease resistance mechanisms interact with non-biotrophic (hemibiotrophic or necrotrophic) fungal pathogens that thrive on dying host tissue during successful infection, is less clear. Several lines of recent evidence have begun to suggest that these organisms may actually exploit components of plant immunity in order to infect, successfully colonize and reproduce within host tissues. One underlying mechanism for this strategy has been proposed, which has been referred to as effector triggered susceptibility (ETS). This review aims to highlight the complexity of interactions between plant recognition and defense activation towards non-biotrophic pathogens, with particular emphasis on three important fungal diseases of wheat (Triticum aestivum) leaves.     

Experimental transmission and tissue tropism of white spot syndrome virus (WSSV) in two species of lobsters, Panulirus homarus and Panulirus ornatus

The susceptibility of two species of lobsters, Panulirus homarus and Panulirus ornatus to white spot syndrome virus (WSSV) was tested by oral route and intramuscular injection. The results revealed that these lobsters were as highly susceptible as marine shrimp when the WSSV was administered intramuscularly. The WSSV caused 100% mortality in both Panulirus homarus and Panulirus ornatus, at 168 and 120 h, respectively, after intramuscular injection and failed to cause mortality when given orally. The presence of WSSV in moribund lobsters was confirmed by single-step and nested PCR, Western blot, histology, and bioassay test. It was found in eyestalk, gill, head muscle, tail muscle, hemolymph, appendages, and stomach. In lobsters with oral route infection, all tested organs except stomach and head muscle was negative for WSSV by nested PCR at 120 h post-inoculation. The stomach and head muscle was positive by nested PCR at 120 h p.i., but negative at 168 h p.i. Western blot analysis was negative in all the tested organs of both species of lobster at 120 h post-inoculation by oral route.     

In Vitro Cultivation of Cymatocarpus solearis (Brachycoeliidae) Metacercariae to Obtain the Adult Stage without the Marine Turtle Definitive Host

In vitro cultivation of trematodes would assist studies on the basic biology of the parasites and their hosts. This is the first study to use the yolk of unfertilized chicken eggs as a simple and successful method of ovocultivation and the first time to obtain the adult-stage of the trematode Cymatocarpus solearis Braun, 1899 (Digenea: Brachycoeliidae). Chicken eggs were inoculated with metacercariae from the muscle of the spiny lobster, Panulirus argus (Latreille, 1804). The metacercariae were excysted and incubated for 576 hr (24 days) at 38℃ to obtain the adult stage. Eggs in utero were normal in shape and light brown color. The metacercariae developed into mature parasites that have been identified as the adult-stage found in marine turtles. The adult lobsters collected in Quintana Roo State, Mexico, showed the prevalence of 49.4% and the mean intensity of 26.0 per host (n = 87). A statistical study was performed to determine that no parasitic preference was detected for male versus female parasitized lobsters. Morphometric measurements of the adult-stage of C. solearis obtained in our study have been deposited in the National Helminths Collection of the Institute of Biology of the National Autonomous University of Mexico. This study is significant because it is the first time that a digenean of the family Brachycoeliidae has been demonstrated to develop in vitro from metacercariae into adults capable of producing eggs using the yolk of unfertilized chicken eggs. Secondly, this technique allows to obtain the adult stage of C. solearis without the presence of its marine turtle host, allows us to describe the mature parasites, and thus contribute to our understanding of the biology of C. solearis.     

In Memoriam: Professor Walter James Harman, PhD (1928–2002)

The Mediterranean spiny lobsters, the common spiny lobster Palinurus elephas (Fabricius, 1758) and the pink spiny lobster P. mauritanicus Gruvel, 1911, are important target species for commercial fisheries. In this study, we focus our attention on the DNA sequence variation of the mitochondrial cytochrome oxydase I gene (COI) in the two species of Palinurus. Spiny lobster DNA samples from four Mediterranean localities were analysed to examine the genetic variability at both the intra- and interspecific level. Furthermore, the phylogenetic relationships within the family Palinuridae (among the two species of Palinurus, most of the species of Panulirus and all the species of Jasus) are examined.