Genetic signature analysis of Perkinsus marinus in Mexico suggests possible translocation from the Atlantic Ocean to the Pacific coast of Mexico.

BACKGROUND: The protozoan Perkinsus marinus (Mackin, Owen & Collier) Levine, 1978 causes perkinsosis in the American oyster Crassostrea virginica Gmelin, 1791. This pathogen is present in cultured C. virginica from the Gulf of Mexico and has been reported recently in Saccostrea palmula (Carpenter, 1857), Crassostrea corteziensis (Hertlein, 1951) and Crassostrea gigas (Thunberg, 1793) from the Mexican Pacific coast. Transportation of fresh oysters for human consumption and repopulation could be implicated in the transmission and dissemination of this parasite across the Mexican Pacific coast. The aim of this study was two-fold. First, we evaluated the P. marinus infection parameters by PCR and RFTM (Ray's fluid thioglycollate medium) in C. virginica from four major lagoons (Términos Lagoon, Campeche; Carmen-Pajonal-Machona Lagoon complex, Tabasco; Mandinga Lagoon, Veracruz; and La Pesca Lagoon, Tamaulipas) from the Gulf of Mexico. Secondly, we used DNA sequence analyses of the ribosomal non-transcribed spacer (rNTS) region of P. marinus to determine the possible translocation of this species from the Gulf of Mexico to the Mexican Pacific coast. RESULTS: Perkinsus marinus prevalence by PCR was 57.7% (338 out of 586 oysters) and 38.2% (224 out of 586 oysters) by RFTM. The highest prevalence was observed in the Carmen-Pajonal-Machona Lagoon complex in the state of Tabasco (73% by PCR and 58% by RFTM) and the estimated weighted prevalence (WP) was less than 1.0 in the four lagoons. Ten unique rDNA-NTS sequences of P. marinus [termed herein the "P. marinus (Pm) haplotype"] were identified in the Gulf of Mexico sample. They shared 96-100% similarity with 18 rDNA-NTS sequences from the GenBank database which were derived from 16 Mexican Pacific coast infections and two sequences from the USA. The phylogenetic tree and the haplotype network showed that the P. marinus rDNA-NTS sequences from Mexico were distant from the rDNA-NTS sequences of P. marinus reported from the USA. The ten rDNA-NTS sequences described herein were restricted to specific locations displaying different geographical connections within the Gulf of Mexico; the Carmen-Pajonal-Machona Pm1 haplotype from the state of Tabasco shared a cluster with P. marinus isolates reported from the Mexican Pacific coast. CONCLUSIONS: The rDNA-NTS sequences of P. marinus from the state of Tabasco shared high similarity with the reference rDNA-NTS sequences from the Mexican Pacific coast. The high similarity suggests a transfer of oysters infected with P. marinus from the Mexican part of the Gulf of Mexico into the Mexican Pacific coast.

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.

Survey of protozoan, helminth and viral infections in shrimp Litopenaeus setiferus and prawn Macrobrachium acanthurus native to the Jamapa River region, Mexico.

We surveyed protozoan and metazoan parasites as well as white spot syndrome virus (WSSV) and infectious hypodermal hematopoietic necrosis virus (IHHNV) in white shrimp Litopenaeus setiferus and the palaemonid prawn Macrobrachium acanthurus native to the lower Jamapa River region of Veracruz, Mexico. The presence of parasites and the infection parameters were evaluated in 113 palaemonid prawns collected during the northwind (n = 45), rainy (n = 38) and dry seasons (n = 30) between October 2007 and July 2008, and in 91 shrimp collected in the rainy season between May and June 2008. In L. setiferus, ciliates of the subclass Apostomatia (Ascophrys sp.) were evident in gills, and third-stage larvae of the nematode Physocephalus sexalatus were evident in the stomach. Cestodes of the genus Prochristianella were evident in the hepatopancreas, while some gregarines of the genus Nematopsis, as well as unidentified larval cestodes, were observed in the intestine. Histology identified Ascophrys sp. in association with gill necrosis and tissue melanization. Slight inflammation was observed in intestinal epithelium near cestode larvae. In M. acanthurus, epibionts of the protozoans Epistylis sp., Acineta sp. and Lagenophrys sp. were observed under uropods, periopods and pleopods. An unidentified ciliate of the Apostomatia was also found in the gills, and Nematopsis was identified in the intestine. No histopathology was observed in association with these parasites. Moreover, neither WSSV nor IHHNV were detected by the polymerase chain reaction (PCR) in any of the L. setiferus or M. acanthurus analysed.

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.

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.

Parasite survey of the eastern oyster Crassostrea virginica in coastal lagoons of the southern Gulf of Mexico.

A parasitological study of the eastern oyster Crassostrea virginica from 11 coastal lagoons in the southern Gulf of Mexico in dry and rainy seasons between late 1999 and early 2001 revealed the presence of 36 bacterial, 2 protozoan (Nematopsis prytherchi and Perkinsus marinus), and 4 helminth species (Urastoma cyprinae, Proctoeces maculatus, a Bucephalus sp., and a Tylocephalum sp.). The prevalence and mean abundances for the protozoa and helminths varied widely between locations but were generally below 50%. Nematopsis prytherchi and the Tylocephalum sp. were the most prevalent species (values were above 60% in most locations). Perkinsus marinus was present in oysters of eight of the coastal lagoons and had low prevalence (<30%) in almost all samples. All identified protozoa and helminths are widely distributed in the Gulf of Mexico and are common oyster parasites. Only P. marinus and the Bucephalus sp. were associated with damage to host tissues. In addition to these parasites, Rickettsia-like bacteria were found in the digestive gland and gills and viral gametocytic hypertrophy inclusions in the gonads by histological examination.

An aromatase inhibitor and tamoxifen decrease plasma levels of o,p'-DDT and its metabolites in Nile tilapia (Oreochromis niloticus).

The objective of this study was to determine if tamoxifen or an aromatase inhibitor (4-hydroxyandrostenedione; 4-OHA) affected plasma concentrations of o,p' -DDT and its metabolites, o,p'-DDD and o,p'-DDE, in mature tilapia. Male and female tilapia were injected 6 times intraperitoneally with o,p'-DDT (40 mg/kg) alone or in combination with 4-OHA (2 mg/kg) over a 12 day period. An additional group of male fish was injected with tamoxifen (5 mg/kg) plus o,p'-DDT. At the end of the treatment period, plasma samples were extracted and analyzed by GC/ECD. Females injected only with o,p'-DDT had significantly higher levels of o,p'-DDT compared with males. Interestingly, females and males treated concomitantly with o,p'-DDT and 4-OHA or tamoxifen had significantly lower concentrations of plasma o,p'-DDT (about 50%) compared with fish treated with only o,p'-DDT. These initial results suggest that an interaction between endocrine-active compounds occurs in vivo in tilapia and may involve alterations in metabolism of o,p'-DDT.

O,p'-DDT induction of vitellogenesis and its inhibition by tamoxifen in Nile tilapia (Oreochromis niloticus).

In order to investigate the mechanism by which o,p'-DDT disrupts endocrine functioning of Nile tilapia in vivo, the estrogenicity of o,p'-DDT was investigated in conjunction with 17beta-estradiol (E2) and tamoxifen. Mature, male tilapia were treated intraperitoneally with o,p'-DDT (60 mg/kg, one dose) or E2 (5 mg/kg, four doses) in the presence or absence of tamoxifen (5 mg/kg, six doses) for 12 days and then plasma vitellogenin (Vtg) (measured as alkaline-labile phosphorous), E2, and testosterone (T) were measured. Vtg levels were increased dramatically by E2 (1,744 +/- 171 microg/ml) and moderately by o,p'-DDT (82 +/- 15 microg/ml) compared with controls (23 +/- 3.5 microg/ml). Tamoxifen alone had no effect on Vtg production, but inhibited both E2 and o,p'-DDT stimulated vitellogenesis. T levels were reduced with E2 administration (1,688 +/- 383 pg/ml) and declined further with the combined treatment of E2 and tamoxifen (281 +/- 70 pg/ml), compared with controls (6,558 +/- 1,438 pg/ml). Tamoxifen or o,p'-DDT alone did not affect T levels, but their combined treatment did (2,069 +/- 647 pg/ml). The results of this study suggest that o,p'-DDT is weakly estrogenic in male tilapia, and that this activity may be mediated through the estrogen receptor.