José Luis Ochoa

Sea bird mortality at Cabo San Lucas, Mexico: Evidence that toxic diatom blooms are spreading

Domoic acid was found to be responsible for an isolated event involving the massive death of brown pelicans (Pelecanus occidentalis) in January 1996, at the tip of the Baja California peninsula. The death of these sea birds was the result of feeding on mackerel (Scomber japonicus) contaminated by domoic acid-producing diatoms (Pseudonitzschia sp.). The number of dead birds (150 animals) found during a period of 5 days caused alarm and called for a governmental task force that would help to implement emergency measurements to protect other species of bird. Also, local canneries were inspected to verify the safety of their recent production and prevent the toxin entering the human market. Fortunately, the timing, response and coordination of this task force enabled identification of the origin and nature of the toxin that provoked such a phenomenon. Future monitoring is recommended to avoid a larger impact of domoic acid spreading and the occurrence of similar toxic events.

Isolation of Prorocentrum lima (Syn. Exuviaella lima) and diarrhetic shellfish poisoning (DSP) risk assessment in the Gulf of California, Mexico

A benthic toxic dinoflagellate identified as Prorocentrum lima (Syn. Exuviaella lima), and designated as strain PRL-1, was isolated from the coast of El Pardito (Coyote) Island in Baja California Sur, Mexico, after a fisherman poisoning incident involving consumption of liver from Lutjanus colorado, and Mycteroperca prionura fish. Purification and culturing was done in ES-Si medium, under 12:12 light/dark cycle (4 x 20 W cool-white fluorescent lamps), at 22 degrees C and constant stirring during 28 days. Whole cells were toxic to Artemia franciscana and its methanolic extract to mouse and to the marine yeast Debaryomyces hansenii. Chromatographic analysis (TLC and HPLC-MS) of such extract indicated an unusual proportion (1:2) okadaic acid (OA) and dinophysistoxin-1 (DTX-1). Estimated total toxin content by mouse bioassay (based on OA toxicity) was 19 pg/cell, a value significantly higher than that found by HPLC-MS (about 5.2 pg/cell, taking into account OA and DTX-1 only), suggesting that additional toxic components of unidentified nature are detected with the bioassay. This is the first report of a successful isolation and culturing of a toxic dinoflagellate from the Gulf of California, Mexico.

Toxic and harmful marine phytoplankton and microalgae (HABs) in Mexican Coasts.

Harmful Algal Blooms (HABs) are becoming an increasing problem to human health and environment (including effects on natural and cultured resources, tourism and ecosystems) all over the world. In Mexico a number of human fatalities and important economic losses have occurred in the last 30 years because of these events. There are about 70 species of planktonic and non-planktonic microalgae considered harmful in Mexican coasts. The most important toxin-producing species are the dinoflagellates Gymnodinium catenatum and Pyrodinium bahamense var. compressum, in the Mexican Pacific, and Karenia brevis in the Gulf of Mexico, and consequently the poisonings documented in Mexico are Paralytic Shellfish Poisoning (PSP) and Neurotoxic Shellfish Poisoning (NSP). Although there is evidence that Amnesic Shellfish Poisoning (ASP), Diarrhetic Shellfish Poisoning (DSP) and Ciguatera Fish Poisoning (CFP) also occur in Mexico, these problems are reported less frequently. The type of phytoplankton and epiphytic microalgae, their toxins and harmful effects as well as current methodology used to study these phenomena are presented in this paper. As an experienced group of workers, we include descriptions of monitoring and mitigation programs, our proposals for collaborative projects and perspectives on future research.

Toxicities and distribution of tetrodotoxin in the tissues of puffer fish found in the coast of the Baja California Peninsula, Mexico.

Toxicities and tetrodotoxin distribution in tissues of five puffer fish species commonly found in the littoral of Baja California Peninsula, Mexico (Sphoeroides annulatus, S. lobatus, S. lispus, Arothron meleagris and Canthigaster punctatissima) were evaluated by bioassay and HPLC. The toxicities estimated as tetrodotoxin-equivalents of all species were more than 0.42 microg/g in at least one of the tissues tested, and the highest was found in S. lispus liver (130 microg/g).

Anti-Inflammatory Activity of Debaryomyces hansenii Cu,Zn-SOD

BACKGROUND Cu,Zn-superoxide-dismutase, Cu,Zn-SOD, can be obtained from different sources with different anti-inflammatory activities. In this study we compared the anti-inflammatory capacity of the marine yeast Debaryomyces hanseii Cu,Zn-SOD (Dh-SOD) with that of bovine erythrocytes (Be-SOD) in a preventive and a therapeutic fashion. METHODS Edema was induced by carrageenan injection into the rat hind paw and was evaluated using a mercury plethysmograph. Development of the inflammatory process was followed by volume displacement at time 0 (carrageenan injection), 1, 2, 3, 4, 5, 6, 9, 12, and 24 h thereafter. Three different SOD doses were used in preliminary experiments to prevent edema: 10, 100, and 1,000 U/kg. RESULTS The results indicate that, at the lowest dose (10 U/kg), both SOD samples are effective in reducing inflammation in both the prostaglandin and amplification phases (-24.8% and -17.5% in the case of Be-SOD, and 11.8% and -18.7% in the case of Dh-SOD, respectively) (p < 0.05). At 100 U/kg, Be-SOD also shows good anti-inflammatory activity in all edema phase (-27.1% in the serotonin phase; -19.4% in the prostaglandin phase; and -20% in the amplification phase) (p < 0.05), but Dh-SOD was less effective (-10.9%, -9.1%, and -5.7%). At the highest dose tested (1000 U/kg), Dh-SOD was, again, more effective than Be-SOD in all three edema phases (-33.1% and -1.5%; -17.9% and -2.6%; and -13.8% and 6.7%, respectively) (p < 0.05). When evaluated as a therapeutic alternative, single doses of Dh-SOD at 1,000 U/kg, and Be-SOD at 100 U/kg, both showed good anti-inflammatory activities (-31.7% and -23.5%, respectively) (p < 0.05). CONCLUSION For therapy purposes alone, Dh-SOD appears to be a better anti-inflammatory agent than Be-SOD in carrageenan-induced edema.

Toxic events in the Northwest Pacific coastline of Mexico during 1992–1995: origin and impact

Previously considered as toxin-free, the Baja California Peninsula has witnessed several toxic algal blooms during the past three years. Apparently these ‘red-tide’ phenomenas outbreaks are not linked to any human related activity. This may just reflect better detection and training. Such events may be periodical and natural rather than induced. The most common types of marine toxins have been detected along the coast of the Peninsula and neighboring waters by mouse bioassay and chromatographic techniques. These are: Tetrodotoxin (TTX), Amnesic Shellfish Poison (ASP), Paralytic Shellfish Poisons (PSP), Diarrhetic Shellfish Poisons (DSP) and even Ciguatera (CFP), which are related to the presence of organisms of Prorocentrum sp. and Alexandrium sp. groups, and the diatom Pseudonitzschia sp. among others. There are also some indications about different kinds of TTX in the puffer fish of the region, and reasons to believe that we are facing a quite different pattern in toxic components, since PSP toxic potency (defined as the number of mouse units per gram(MU/g)of shellfish meat) is very high in spite of low dinoflagellates cell density registered. The ecological and social impact of the above has been considerable, with mass deaths of shellfish, seagulls, dolphins and turtles, and even some human casualties. The locally registered toxicity records: PSP found in one single fanshell reaches to 23 000 MU/100 g of tissue as determined by the mouse bioassay and, on a different event, two persons killed after ingesting puffer fish fillet. The largest reservoir of commercial marine organisms in Mexico is precisely the Northwest coast of the country and important plans for building large harbors and develop aquaculture areas are in progress. Therefore, a monitoring program is essential for an adequate management of such resources. Considering the large extension of the Peninsula (about1600 km)and, at this time, the lack of efficient communication means and scarce population, the implementation of such monitoring programs presents a big challenge.

Cloning and sequencing of a cDNA encoding a copper-zinc superoxide dismutase enzyme from the marine yeast Debaryomyces hansenii

Cu‐Zn superoxide dismutase (SOD‐1) is a ubiquitously occurring eukaryotic enzyme with a variety of important effects on respiring organisms. A gene (dhsod‐1) encoding a Cu‐Zn superoxide dismutase of the marine yeast Debaryomyces hansenii was cloned using mRNA by the RT‐PCR technique. The deduced amino‐acid sequence shows ∼70% homology with that of cytosolic superoxide dismutase from Saccharomyces cerevisiae and Neurospora crassa, as well as lower homologies (between 55 and 65%) with the corresponding enzyme of other eukaryotic organisms, including human. The gene sequence encodes a protein of 153 amino acids with a calculated molecular mass of 15·92 kDa, in agreement with the observed characteristics of the purified protein from D. hansenii. The dhsod‐1 sequence has been deposited in the public data library of the NCBI under Accession Number AFO 16383.

Retention and tissue damage of PSP and NSP toxins in shrimp: Is cultured shrimp a potential vector of toxins to human population?

Toxic microalgae outbreaks have caused significant economic losses in the Mexican aquaculture industry. Blooms that involve PSP and NSP phycotoxins are two of the most dangerous, causing harmful effects to the environment, economy and public health. The exact metabolic mechanism of these toxins in shrimp still remains unknown. Because shrimp consume microalgae their edible tissues are clearly possible vectors for human toxic syndrome. This study examined and verified the toxicological effects for white leg shrimp (Litopenaeus vannamei) exposed to different cell densities of Gymnodinium catenatum and Karenia brevis. Acute assays demonstrated good survival rates of shrimp at low densities of dinoflagellates (10(3) cell/L), while mortality and abnormal behavior were observed with higher densities (>10(4) cell/L). Chronic assays showed significant differences in survival rates, percentage of feed and weight gain of organisms exposed to the dinoflagellates with respect to controls. Furthermore, PSP and NSP toxins were detected in all the edible tissues. Gastric glands and muscle retained toxins for a longer period of time compared to other tissues, even after a depuration period. Histology damages were observed in the heart, gastric gland and brain. This study strongly supports that shrimp represent a potential risk for humans as unconventional vectors of phycotoxins.

Distribution of Sporobolomyces (Kluyver et van Niel) genus in the western coast of Baja California Sur, Mexico

Summary One hundred forty one strains of yeast were isolated from 98 seawater samples collected in the Pacific Ocean, off the west coast of Baja California, Mexico. The genera Sporobolomyces represented 32% of the total isolates, and was comprised of the species Sp. holsaticus (10%), Sp. puniceus (20%) and Sp. roseus (2%). By correlating the distribution of marine yeast with temperature, salinity, dissolved oxygen and depth, it was possible to establish the distribution pattern for each species.

Effect of Superoxide Dismutase From Bovine Erythrocytes on Different Activity Parameters in Adjuvant-Induced Arthritis

BACKGROUND The purpose of this work was to evaluate the effect of superoxide dismutase (SOD) on primary swelling, lipoperoxidation, body thymus, and spleen weight in the adjuvant-induced arthritis (AIA) model in rats. METHODS Orally and intraperitoneally administered SOD (100 U/kg) from bovine erythrocytes, as well as naproxen (40 mg/kg) and dexamethasone (25 mg/kg), were evaluated against placebo. RESULTS Primary edema was not decreased by SOD; in contrast, naproxen and dexamethasone showed good anti-inflammatory activity. Lipoperoxidation increased 1.8, 2.5, and 2.8 times with intraperitoneal SOD, naproxen, and dexamethasone administration, respectively, while oral SOD decreased lipoperoxidation levels to approximately one-half of that found in the control group. Body weight increased with SOD but decreased with dexamethasone. Naproxen did not change the animal weight. Thymus weight remained unchanged with SOD and naproxen, while it decreased with dexamethasone. Spleen weight remained the same with SOD, but increased with naproxen and decreased with dexamethasone. No side effects were observed in the SOD group, whereas 20% of the rats in the naproxen group died of gastrointestinal hemorrhage, and 50% of the rats in the dexamethasone group, of pulmonary infection. CONCLUSIONS In conclusion, SOD showed no anti-inflammatory activity but decreased lipoperoxidation when administered orally. No deleterious effects in primary and secondary immunologic organs were observed with this agent.