In the spiny lobster (Panulirus interruptus) the prophenoloxidase is located in plasma not in haemocytes

Hernández‐López, Jorge; Gollas‐Galván, Teresa; Gómez‐Jiménez, Silvia; Portillo-Clark, Guillermo; Vargas‐Albores, Francisco

doi:10.1006/fsim.2002.0419

Cited by 28 publications

(21 citation statements)

References 39 publications

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“…Lobster hemolymph without anticoagulants and the C: 500 bp marker anticoagulants themselves can inhibit the PCR reaction. We used 2 anticoagulants successfully in the present study: sodium citrate/EDTA (Söderhäll & Smith 1983) and lobster salt solution with EDTA (Hernández-López et al 2003). Both inhibit PCR, likely due to their EDTA content, but both anticoagulants can be removed from the sample DNA in the extraction process.…”

Section: Discussionmentioning

confidence: 99%

Detection of Panulirus argus Virus 1 in Caribbean spiny lobsters

Montgomery-Fullerton¹,

Cooper²,

Kauffman³

et al. 2007

Dis. Aquat. Org.

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Panulirus argus Virus 1 (PaV1) is a pathogenic virus that infects Caribbean spiny lobsters P. argus in the Florida Keys. We have developed a PCR detection assay for PaV1 for the purpose of studying the natural history of the virus and for monitoring the prevalence of infection. The detection of the virus in hemolymph and other tissues is based on the PCR amplification of a 499 bp product using specific primers designed from a cloned fragment of the PaV1 genome. The sensitivity limit for the assay was 1.2 fg of purified viral DNA. The PaV1 primers did not react with lobster DNA, oyster DNA, Ostreid Herpesvirus 1, or murine cytomegalovirus. Using this assay, we successfully followed the course of infection in lobsters inoculated with PaV1 and we detected infections in wild-caught lobsters from the Florida Keys. We have also established guidelines for interpreting infection results from the PCR assay for PaV1. KEY WORDS: Assay · Lobster virus · Panulirus argus · Molecular detection Resale or republication not permitted without written consent of the publisherDis Aquat Org 76: [1][2][3][4][5][6] 2007 MATERIALS AND METHODS Lobsters. Juvenile spiny lobsters Panulirus arguswere collected in the Florida Keys from shallow (1 to 2 m) sub-tidal habitats, by hand using SCUBA, and transported in seawater-filled coolers to the Keys Marine Laboratory, Long Key, Florida, USA. Lobsters were shipped overnight to the Virginia Institute of Marine Science, Virginia, USA, where they were held in 78 l aquaria equipped with aerators and 2 Whisper filters containing preconditioned crushed coral. Artificial seawater (Marinemix Forty Fathoms, Marine Enterprises International) in the aquaria was monitored weekly for pH (7.1 to 7.6), salinity (33 to 35 ppt), temperature (21 to 24°C), ammonia (<1.0 ppm), and nitrite (<1.0 ppm), and 50% water changes were performed to maintain water quality within acceptable limits. Lobsters were fed squid 3 times per week, and any food remaining in aquaria after 3 h was removed. Healthy individuals have been maintained in this system for > 2 yr. Diseased and healthy animals, distinguished by the presence of discolored hemolymph and obvious signs of morbidity, were housed in separate aquaria in different rooms. The virus has been maintained for > 3 yr in the laboratory through repeated passages of infected hemolymph into healthy animals.All hemolymph samples from healthy, naturally infected, and inoculated lobsters were collected in an equal volume of citrate anticoagulant (Söderhäll & Smith 1983) and frozen at -80°C until tested. The stock anticoagulant solution also contained 20% dimethyl sulfoxide (Sigma-Aldrich), 0.8% protease inhibitor cocktail (Sigma-Aldrich P8340), and 10 IU bacitracin ml -1 (Fisher Scientific). For inoculation trials, pre-inoculation hemolymph was collected from each lobster immediately prior to needle inoculation with 0.1 ml of pooled, infected hemolymph from 3 diseased lobsters mixed 1:1 in anticoagulant. In preliminary tests, the injection of our stock anticoagulant s...

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Section: Discussionmentioning

confidence: 99%

Detection of Panulirus argus Virus 1 in Caribbean spiny lobsters

Montgomery-Fullerton¹,

Cooper²,

Kauffman³

et al. 2007

Dis. Aquat. Org.

View full text Add to dashboard Cite

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“…1A). The significant reduction in THC at salinity 45& could be associated with the significant decrease in granular cells as the proPO activating enzyme (PPAE) has to be released from the haemocyte granules, which has been suggested as the natural activation pathway in response to stress [19,27]. In addition, increase in PO activity could also be due to significant reduction in plasma inhibitors regulating the proPO system [18,27].…”

Section: Discussionmentioning

confidence: 99%

“…Although in the spiny lobster (Genus: Panulirus) the proPO activating mechanism is similar to other crustaceans, unlike other crustaceans, most of the proPO-activity of spiny lobster has been detected in cell-free plasma [19]. In addition, Perazzolo et al [20] reported a significant level of PO activity in the serum of Penaeus paulensis suggesting the use of serum for PO activity assay.…”

Section: Phenoloxidase (Po) Assaymentioning

confidence: 99%

Effect of environmental parameters on immune response of the Indian spiny lobster, Panulirus homarus (Linnaeus, 1758)

Verghese

Radhakrishnan

Padhi

2007

Fish & Shellfish Immunology

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“…Hemolymph was diluted (1:3) with pre-chilled (8°C) anticoagulant according to Hernández-López et al (2003), who titrated the appropriate concentration of NaCl to avoid lysis of hemocytes in Panulirus argus (350 mM NaCl, 10 mM KCl, 10 mM HEPES, 10 mM EDTA-Na 2 , pH 7.3, 850 mOsm kg −1 ). The sample was then centrifuged at 800 × g for 5 min at 4°C to separate the plasma, which was used to evaluate plasmatic metabolites, phenoloxidase (PO) activity, hemagglutination activity, and trypsininhibitory ac tivity.…”

Section: Plasma and Degranulated Hemocyte Preparationmentioning

confidence: 99%

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

Pascual-Jímenez¹,

Huchin-Mian²,

Simões³

et al. 2012

Dis. Aquat. Org.

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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.

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In the spiny lobster (Panulirus interruptus) the prophenoloxidase is located in plasma not in haemocytes

Cited by 28 publications

References 39 publications

Detection of Panulirus argus Virus 1 in Caribbean spiny lobsters

Detection of Panulirus argus Virus 1 in Caribbean spiny lobsters

Effect of environmental parameters on immune response of the Indian spiny lobster, Panulirus homarus (Linnaeus, 1758)

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

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