Association between endotoxin levels in dust from indoor swine housing environments and the immune responses of pigs

Indoor animal husbandry environments are inevitably contaminated with endotoxins. Endotoxin exposure is associated with various inflammatory illnesses in animals. This cross-sectional study evaluated the relationship between the degree of endotoxin exposure and the cellular and humoral immune profiles of fattening pigs. Blood samples were taken from the jugular vein of 47 pigs from ten pig farms in Korea. Whole blood cell counts and plasma immunoglobulin (Ig) classes were determined. Peripheral-blood mononuclear cells were stimulated in vitro with concanavalin A for 48 h, and cytokines released into culture supernatants were measured. The barns in which the pigs lived were assessed for endotoxin levels in the total and respirable dust by using the limulus amebocyte lysate kinetic QCL method. Low and high endotoxin exposures were defined as ≤ 30 and > 30 EU/m³, respectively. Compared to pigs with low endotoxin exposure (n = 19), highly exposed pigs (n = 28) had higher circulating neutrophil and lymphocyte (particularly B cells) counts, IgG and IgE levels, interferon-gamma (IFNγ) and interleukin (IL)-4 productions, and lower IgA levels and tumor necrosis factor-alpha (TNFα) production. The IL-4, IFNγ, and TNFα levels significantly correlated with endotoxin level and/or pig age. Constant exposure of pigs to high levels of airborne endotoxins can lead to aberrant immune profiles.

Epizootiological characteristics of viable bacteria and fungi in indoor air from porcine, chicken, or bovine husbandry confinement buildings

Microorganisms found in bioaerosols from animal confinement buildings not only foster the risk of spreading diseases among livestock buildings, but also pose health hazards to farm workers and nearby residents. This study identified the various microorganisms present in the air of swine, chicken, and cattle farms with different kinds of ventilation conditions in Korea. Microbial air samples were collected onto Petri dishes with bacterial or fungal growth media using a cascade impactor. Endotoxin levels in total dust were determined by the limulus amebocyte lysate kinetic QCL method. Prevalent Gram-positive bacteria were Staphylococcus (S.) lentus, S. chromogenes, Bacillus (B.) cereus, B. licheniformis, and Enterococcus faecalis, while the dominant fungi and Gram-negative bacteria were Candida albicans and Sphingomonas paucimobilis, respectively. Considering no significant relationship between the indoor dust endotoxin levels and the isolation of Gram-negative bacteria from the indoor air, monitoring the indoor airborne endotoxin level was found to be also critical for risk assessment on health for animals or workers. The present study confirms the importance of microbiological monitoring and control on animal husbandry indoor air to ensure animal and worker welfare.

Effect of perivitelline fluid from horseshoe crab on the expression of COL1A1 in dental pulp stem cells

Perivitelline fluid, extracted from the fertilized eggs of horseshoe crabs, has been reported to play a vital role in supporting embryogenesis as well as cell proliferation. The present study aims to evaluate the effect of PVF on the expression of COL1A1 in human dental pulp stem cells (DPSCs). The cells were grouped into two; untreated (control) and treated with a single dose of PVF (0.019 mg/ml). Gene expression was quantified for COL1A1 on day 1, 3 and 7 using reverse transcriptase PCR. The expression of COL1A1 on day 3 of treated group with PVF was the highest though there was a decline of COL1A1 expression on day 7. Mann Whitney test was utilized to determine the significance of COL1A1 expression between treated and untreated groups. Significant difference in the expression of COL1A1 was observed between the treated and untreated groups on day 3 though there was no significance in the expression on day 7. The present study indicates that PVF may have the potential to increase cell proliferation in human DPSCs.

Population structure of the red mangrove crab, Goniopsis cruentata (Decapoda: Grapsidae) under different fishery impacts: Implications for resource management / Estructura de la población del cangrejo rojo de manglar, Goniopsis cruentata (Decapoda: Grapsidae) bajo diferentes impactos de pesca: Implicaciones para la gestión de los recursos

The red mangrove crab, Goniopsis cruentata, influences the recruitment and composition of plant species in the mangrove ecosystem and it is an important fishery resource. Nevertheless, no current management and conservation plans are available for this species for the Brazilian coast. This investigation evaluated the population structure and reproductive biology in populations of G. cruentata under contrasting fishery pressures. The sampling program was carried out in two mangroves, Vaza-Barris and Sergipe River, from January through December 2011. Crabs from both mangroves were randomly collected by a professional fisherman during daytime low tide periods, using a fishing rod baited with pieces of a locally abundant gastropod, Pugilina morio, during 20min/area (catch per unit effort). Monthly measurements of air, sediment surface layer and water temperatures were obtained with a digital thermometer and salinity with an optical refractometer. Both crab populations were compared concerning their abundance, body size, sex ratio, size at onset of sexual maturity and fecundity (FI). Abiotic factors (air, water and mud temperature; and salinity) showed no significant differences between sampling localities. A total of 4 370 crabs were sampled, 2 829 from the Sergipe River and 1 541 from the Vaza-Barris River. The abundance and body size of crabs were compared between mangroves, and statistically significant differences were found. The sex ratio for both populations differed from the expected 1:1 ratio, and a significant deviation in favor of juvenile males was obtained, while adults showed a bias toward females. The estimated size at onset of sexual maturity for both sexes was similar in both populations. However, the populations differed significantly in the number and volume of eggs: a higher FI was obtained in females from the Sergipe River, while a higher egg volume was observed in females from the Vaza-Barris River mangrove. These results indicated a tendency to decrease the body size, the abundance of crabs and the reproductive potential of the species, with higher fishing intensities, reinforcing the need to develop a management plan as well as to establish conservation units for G. cruentata in the Northeastern Brazilian coast. Habitat loss in conjunction with long-term overfishing can have irreversible consequences, which can impact not only the populations of commercially exploited crabs, but the dynamics of virtually the entire mangrove ecosystem.

El cangrejo rojo Goniopsis cruentata influye en el reclutamiento y la composición de especies vegetales del ecosistema manglar y es un importante recurso pesquero. Sin embargo hasta la fecha, no se han desarrollado planes de manejo y conservación para esta especie en Brasil. La presente investigación evaluó la estructura poblacional y la biología reproductiva de las poblaciones de G. cruentata expuestas a diferentes presiones de pesca. El muestreo se realizó en dos manglares, Río Sergipe y Río Vaza-Barris entre enero y diciembre 2011. En ambos manglares, los animales fueron recolectados de día durante la marea baja por un pescador profesional, utilizando una caña de pescar con carnada del gasterópodo Pugilina morio, durante 20min/área (captura por unidad de esfuerzo, CPUE). Mensualmente con un termómetro digital se registró la temperatura del aire, agua y capa superficial del sedimento, mientras que la salinidad fue cuantificada utilizando un refractómetro óptico. Las poblaciones fueron comparadas en su abundancia, tamaño corporal, proporción de sexos, talla de primera madurez sexual y fecundidad. Los factores abióticos(temperatura del aire, agua y sedimento y la salinidad) no mostraron diferencias significativas entre ambos sitios de muestreo. Se recolectaron un total de 4 370 cangrejos; 2 829 en el río Sergipe y 1 541 en Vaza-Barris. Se registraron diferencias significativas entre los manglares con respecto a la abundancia y el tamaño corporal de los animales. La proporción de sexos de ambas poblaciones difirió de la relación 1:1 esperada y se obtuvo una desviación significativa en favor de machos jóvenes, mientras que los adultos mostraron una tendencia hacia las hembras. A su vez, las poblaciones difirieron significativamente en el número de huevos, con un mayor IF para las hembras del manglar del río Sergipe. El volumen de los huevos también mostró diferencias significativas entre ambas poblaciones. Estos resultados indican que la alta intensidad de pesca podría estar afectando dichos parámetros, lo que refuerza la necesidad de desarrollar un plan de manejo y la creación de unidades de conservación para G. cruentata en la costa noreste de Brasil. La pérdida del hábitat junto con la sobrepesca a largo plazo, pueden tener consecuencias irreversibles, que afectan no sólo a las poblaciones de cangrejos de importancia comercial, sino también la dinámica del ecosistema de manglar.

Colour preference and light sensitivity in trilobite larvae of mangrove horseshoe crab, Carcinoscopius rotundicauda (Latreille, 1802).

The trilobite larvae of C. rotundicauda were tested to determine their colour preference and light sensitivity until their first moulting (25 days post hatching) under laboratory conditions. Maximum congregation size of the trilobite larvae was found in the white zone respectively where (n= 12) followed by yellow (n= 8) and orange (n= 8), which showed the larval preference for lighter zones. Morisita’s index calculation showed a clumped/aggregated distribution (yellow, blue, orange and white) and uniform/hyper dispersed distribution (green, red and black) for various colours tested. Trilobite larvae showed least preference for brighter regions while tested in the experiment [black; (n=4) and red; (n=5)]. Experiments done to determine the light sensitivity of trilobite larvae showed that the larvae had more preference towards ultraviolet lights. The maximum congregation size of 38.8 and 40.7% of the larvae was encountered under ultraviolet light, when the light sources were kept horizontal and vertical, respectively. Overall, results suggested that the trilobite larvae of C. rotundicauda, preferred light source of shorter wavelengths (UV light) and colours of lighter zone (white, yellow, orange), which might be due to their adaptation to their natural habitat for predator avoidance, prey selection and water quality.

Effects of plasma endotoxin concentrations on the severity of atopic dermatitis in children

PURPOSE: Endotoxin exposure is associated with the development and pathogenesis of allergic diseases. The present study aimed to determine whether plasma endotoxins are elevated in children with atopic dermatitis (AD) and whether the plasma endotoxins are associated with the AD severity. METHODS: This study included 42 children with AD and 16 controls. The AD severity was assessed and skin prick tests were performed along with serum total IgE, total eosinophil count, and ImmunoCAP analyses for common allergens. For plasma endotoxin levels, kinetic turbidimetric assay and limulus amebocyte lysate reagent were used. RESULTS: The endotoxin levels were significantly higher in children with AD than in controls (0.039 [interquartile range {IQR}, 0.031 to 0.046] EU/mL vs. 0.031 [IQR, 0.019 to 0.035] EU/mL, P<0.05). The endotoxin levels showed a significantly inverse correlation with the disease severity in the extrinsic AD (r=-0.414, P=0.017) but not in the intrinsic AD (r=0.222, P=0.426). No significant correlation was observed between plasma endotoxin levels, serum total eosinophil count, and serum total IgE. CONCLUSION: Our results suggest that plasma endotoxin level is increased in the AD subjects than in the normal controls and affects the disease severity only in the extrinsic AD subjects. Further studies with larger sample sizes are warranted to investigate whether plasma endotoxin levels have a different effect on the AD severity depending on the presence of atopy.

A Hyperactive Neutrophil Phenotype in Aggressive Periodontitis

Although neutrophils function in both defense and tissue destruction, their defensive roles have rarely been studied in association with periodontitis. We hypothesized that peripheral neutrophils are pre-activated in vivo in periodontitis and that hyperactive neutrophils would show enhanced phagocytic ability as well as an increased production of reactive oxygen species (ROS). Peripheral blood neutrophils from patients with aggressive periodontitis and age/gender-matched healthy subjects (10 pairs) were isolated. The levels of CD11b and CD64 expression on the neutrophils and the level of plasma endotoxin were determined by flow cytometry and a limulus amebocyte lysate test, respectively. In addition, neutrophils were subjected to a flow cytometric phagocytosis assay and luminol-enhanced chemiluminescence for non-opsonized Fusobacterium nucleatum in parallel. The neutrophilsfrom most patients expressed increased levels of both CD11b and CD64. In addition, the plasma from these patients tended to contain a higher level of endotoxin than the healthy controls. In contrast, no differences were found between the two groups with regard to phagocytosis or ROS generation by F. nucleatum. The ability to phagocytose F. nucleatum was found to positively correlate with the ability to produce ROS. In conclusion, peripheral neutrophils from patients with aggressive periodontitis are hyperactive but not hyperreactive to F. nucleatum.

Content detection of bacterial endotoxin in two kinds of injection by gelatin technique / 中国中药杂志

<p><b>OBJECTIVE</b>To detect content of bacterial endotoxin in Yuxingcao and Qingkailing injections by specific and nonspecific tachypleus amebocyte lysate technique for in order to investigate the feasibility of specific tachypleus amebocyte lysate technique for detecting bacterial endotoxin in traditional Chinese drug injections.</p><p><b>METHOD</b>Different batches of Yuxingcao and Qingkailing injections were detected by specific and nonspecific tachypleus amebocyte lysate kits.</p><p><b>RESULT</b>Yuxingcao injection could be detected by specific and nonspecific tachypleus amebocyte lysate technique, Whereas Qingkailing injection could be detected only by specific tachypleus amebocyte lysate.</p><p><b>CONCLUSION</b>Using specific tachypleus amebocyte lysate as a substitute for nonspecific tachypleus amebocyte lysate is an effective method for detecting content of bacterial endotoxin in Qingkailing injection.</p>

Tetrodotoxin poisoning following ingestion of the toxic eggs of the horseshoe crab Carcinoscorpius rotundicauda, a case series from 1994 through 2006.

Between 1994 and 2006, a total of 280 cases of varying degrees of tetrodotoxin poisoning following ingestion of the toxic eggs of the horseshoe crab, Carcinoscorpius rotundicauda, were admitted to the medical service of Chon Buri Hospital. The severity of the poisoning was classified into four stages based on clinical signs and symptoms of human tetrodotoxication. Of 245 available medical records; 100 were in stage 1, 74 were in stage 2, 3 were in stage 3 and 68 were in stage 4. The frequencies of symptoms and signs included the following : circumoral and lingual numbness (98%), hands and feet numbness (94.7%), weakness (59.6%), dizziness and vertigo (54.3%), nausea and vomiting (52.6%), transient hypertension (39.6%), respiratory paralysis (27.7%), fixed dilated pupils (14.7%), ophthalmoplegia (12.2%), blood pressure lower than 90/60 mmHg (5.7%), and polyuria (0.4%). All patients received symptomatic and supportive treatment. Endotracheal intubation and mechanical ventilation were considered when paralysis was progressing rapidly. The results of treatment are as follows: 239 patients (97.5%) showed complete recovery, 5 patients (2%) died, and 1 patient (0.4%) suffered anoxic brain damage. Horseshoe crab poisoning occurs both sporadically and epidemically in Chon Buri. Seasonal variation in the number of cases of poisoning was observed with a peak from December through March.

The Effect of Preoperative Oral Application of Bovine Colostrum on Endotoxemia and TNF-alpha in Abdominal Surgery

PURPOSE: We conducted this study to assess the effect of oral application of bovine colostrum on the plasma endotoxin and TNF-alpha following the abdominal surgery. METHODS: 46 patients who underwent the abdominal surgery were evenly enrolled in a prospective, randomized, double blind and placebo-controlled study. The patients were preoperatively managed by oral application of 20 g of a bovine colostrums or placebo (nonfat dried milk) per a day for 3 days. In both groups, endotoxin was sequentially determined pre-, intra- and postoperatively by a modified limulus amebocyte lysate test. TNF-alpha and CRP were also measured. The clinical course was followed and compared in both groups. RESULTS: The colostrum group showed significantly lower level of endotoxin and TNF-alpha compared to the placebo group. CRP did not differ in both groups. Clinical event did not occur in both group until the discharge. Colostrum group tends to have a lower incidence of fever and leukocytosis without statistic significance. CONCLUSION: Prophylactic preoperative oral application of bovine colostrum lower the plasma level of perioperative endotoxin and TNF-alpha. Further studies were needed for the relation of clinical effect and preoperative oral application of bovine colostrum.

Epidemic Investigation of Pyrogenic Reactions caused by Contaminated Intravenous Products / 병원감염관리

PURPOSE: This study was conducted to investigate the epidemic pyrogenic reactions that occurred hospital-widely in 51 patients on 30-31 Mar. 1995 at a tertiary, educational, and acute care hospital. METHODS: The remained intravenous (IV) drugs and IV fluids that were infused to patients (N=51) who developed pyrogenic reactions were cultured and tested pyrogenic matter. Pyrogenic test was done by Limulus Amebocyte Lysate (PyrotellRMA,USA). RESULTS: IV normal saline 50 cc bottles (AAXSAT and AAX5AV) those were collected being used from case patients, and new bottles stocked in the drug storage were positive to pyrogenic test. Used normal saline collected from case patients were cultured coagulase-negative Staphylococus. Moreover the company, manufacturer of these fluids reported the positive pyrogenic test on these IV fluids (AAXSAT and AAX5AV). CONCLUSION: This epidemic was a pyrogenic reaction for infused the intrinsic contaminated fluid with pyrogen. We could prevent not the additional pyrogenic reactions in study hospital but national wide epidemics to apply the rapid and well-designed epidemic investigation and infection control.

House Dust Endotoxin Exposure and Allergic Sensitization in Korean Home Living Conditions / 소아알레르기및호흡기학회지

PURPOSE: Household endotoxin exposure in allergy and asthma has been gaining attention for its dual potential to exacerbate these conditions in individuals with established disease and to abrogate atopy before disease onset. The aim of this work was to analyze associations between current exposure to bacterial endotoxin in house dust and allergic sensitization in adults with asthma. METHODS: From the homes of 52 adults with asthma and 28 normal controls, house dust endotoxin (detected with a Limulus Amebocyte Lysate QCL-1000) and house dust mite allergens (Der f 1 and Der p 1) were quantified. Allergen sensitization was measured by skin prick test. RESULTS: The endotoxin levels from mattresses were lower in the home of adults with asthma than in that of normal controls. The endotoxin levels from mattresses detected in the home of adults with asthma were positively correlated with Der p 1 levels and wheal size to housedust mite allergens on skin prick tests. Furthermore, the endotoxin levels in living room floor dust were positively correlated with Der f 1 levels, but didn't relate with wheal size to house dust mite allergens. CONCLUSION: Our results indicate that current exposure to house dust endotoxin might be positively associated with allergic sensitization in adults with asthma.

Effect of bovine colostrum on the bacterial translocation and intestinal endotoxemia in 5-fluorouracil treated rats / 대한내과학회지

BACKGROUND: Antitumor drugs such as 5-fluorouracil (5-FU) are known to induce intestinal damages and bacterial translocation. The present studies examined whether or not bovine colostrum protects against gut barrier damage, bacterial translocation and endotoxemia from these antitumor drugs. METHODS: Rat received either no drug, chemotherapy alone (5-FU, 300 mg/kg intraperitoneal injection) or bovine colostrum (4 g/day per os) for 5 days prior to 5-FU and for 5 days afterward. Intestinal permeability, enteric aerobic bacterial counts, serum albumin and protein levels, and pathologic findings of ileum were measured. Bacterial translocation to systemic blood, mesenteric lymph nodes, liver and spleen were measured. Systemic plasma endotoxin levels were quantified by the chromogenic limulus amebocyte lysate (LAL) technique. RESULTS: 5-FU increase intestinal permeability and plasma endotoxin levels, and decreased serum levels of total protein and albumin. Also 5-FU induced bacterial translocation to mesenteric lymph nodes, liver and spleen, not to systemic blood, but did not induce changes of enteric bacterial numbers and mucosal damages of small intestine. Combined administration of bovine colostrum with 5-FU reduced an increase in intestinal permeability and declines in serum albumin and protein levels by 5-FU. Bovine colostrum supplements also reduced bacterial translocation to mesenteric lymph nodes, liver and spleen, and endotoxemia. CONCLUSION: Bovine colostrums may beneficial effects in preventing 5-FU induced gut barrier damage, bacterial translocation and intestinal endotoxemia.

Influence of immunization dose schemes on the immune response to anti-tetrodotoxin vaccine / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the relationship between the immune response of anti-tetrodotoxin vaccine, including its dose-response, and to select optimal immunization dose so as to enhance antitoxic effect of the anti-tetrodotoxin vaccine.</p><p><b>METHODS</b>Tetrodotoxin (TTX) was coupled to Tachypleus tridentatus hemocyanin (TTH) chemically to form artificial antigen (TTX-TTH), and with which Balb/c mice were immunized. Influence of different immunization doses [100 microg as the higher (H) and 25 microg as the lower (L) dose group] on the protective effects of TTX vaccine was compared. The quality of antisera and effects of vaccine in anti-TTX poisoning were observed.</p><p><b>RESULTS</b>The sera antibody quality increased more quickly in group L than that in group H after immunization. The dose at which the half of immunized mice survived when challenged once with TTX were 16 x LD (1 LD = 13.5 microg/kg, i.p.) in group L and 11 x LD in group H. When TTX was used time and again, the half of immunized mice could tolerate as high as 40 x LD and 22 x LD of accumulated dose, and the maximum tolerable cumulated dose was 104 x LD and 90 x LD for group L and H respectively. The scheme L was better both in antibody quality and effect of protecting against TTX toxicity than that in scheme H.</p><p><b>CONCLUSIONS</b>The experimental vaccine of TTX could effectively protect animal from TTX intoxication. The lower immunization dose in this study is selected as the optimal immunization scheme.</p>

The Effects of Inline Intravenous Filtration on Bacteria, Candida and Bacterial Endotoxin Retentions / 감염

BACKGROUND: In-line intravenous filters have been used in intravenous administration sets of fluid to retain bacteria and bacterial endotoxin. We evaluated the effects of intravenous filters on Staphylococcus epidermidis, Pseudomonas aeruginosa, Candida and bacterial endotoxin retention. METHODS: S. epidermidis, C. albicans, P. aeruginosa and endotoxin were injected into each 10 bags of amino acid solution. After incubation for 24 and 72 hours, aspirates of 0.1 mL from 30 bags, with filtration and without filtration, were inoculated into culture media. The effluent from bags containing endotoxin was tested by limulus amebocyte lysate test. RESULTS: When effluents were filtered before culture, those from each 10 bags containing P. aeruginosa, C. albicans and endotoxin were sterile; also there was only one positive bacterial culture among 10 effluents from bags containing S. epidermidis. But by contrast all effluents without filtration showed positive cultures and endotoxin detection. CONCLUSION: This result showed that inline intravenous filters were useful device to remove bacteria, Candida, and bacterial endotoxin.

The Extraction of Vibrio vulnificus Lipopolysaccharide by the Phenol-Chloroform-Petroleum Ether Method and Biological Activity of the LPS / 대한피부과학회지

BACKGROUND: Vibrio vulnificus is a pathogenic, marine, halophilic, gram-negative bacillus which causes fulminant infecticn in humans through the ingestion of raw seafood or skin wounds. V. vulnificus produces seveal kinds of virulent factors including cytolysin, endotoxin, exoenzymes, and siderphores. Among these, the lipopolysaccharide(LPS) of V. vulnificus has recently been purified, but the biological activity of this endotoxin is not well clarified as yet. OBJECTIVE: The purpose of this study was to extract LPS from Vibrio vulnificus and to test the biological activity of extracted LPS for the elucidation of the role in V. vulnificas septicemia. METHODS: V. vulnificus LPS was extracted by the Phenol Chloroform-Petroleurn ether(PCP) method. The biological activity of LPS was evaluated with a limulus amebocyte lysate assay and by assessment of lethality to ICR mice. RESULTS: Five hundreds mg of LPS were extracted from 10g of dried V. vulnificus. Lirnulus amebocyte lysate formed a hard gel in response to the extracted LPS. This LPS showed low level of protein contaminatior in SDS-PAGE electophoresis and spectrophotometry. A High dose of LPS(200 mg/ml body weight) was lethal to mice. CONCLUSION: The PCP extraction yield relatively large amounts of LPS from V. vnlnificus. with out significant protein contamination and e xtrated LPS has endotoxin activity. This extrated LPS can be used for further studies such as making antibody or characterizing pathogenic roles in the V. vulnificus infection.

An outbreak of horseshoe crab poisoning in Chon Buri, Thailand: clinical, toxicologic and therapeutic considerations.

In 1994-1996, an outbreak of horseshoe crab poisoning by eating toxic eggs of the horseshoe crab Carcinoscorpius rotundicauda affected over 100 persons in Chon Buri which located on the eastern coast of Thailand. We discuss clinical description and management of this first major outbreak. The responsible toxin has been partially purified by means of ultrafiltration and high performance liquid chromatography. The horseshoe crab toxin is identified as tetrodotoxin (TTX) and anhydro TTX.

An epidemic of tetrodotoxin poisoning following ingestion of the horseshoe crab Carcinoscorpius rotundicauda.

At certain seasons of the year in Thailand, the horseshoe crab Carcinoscorpius rotundicauda may be toxic to human and fatal poisoning occasionally occur. Tetrodotoxin (TTX) and its derivatives were major toxins in the toxic eggs of the horseshoe crab. An epidemic of poisoning by eating toxic eggs of the horseshoe crab affected 71 persons in Chon Buri which located in the eastern coast of Thailand. Patients generally presented with neurologic symptoms such as paresthesia, vertigo, weakness, respiratory paralysis, altered consciousness with unreactive dilated pupils in addition to gastrointestinal symptoms such as nausea and vomiting. Nineteen patients required artificial ventilation and there were two deaths. This is the first large outbreak of tetrodotoxin poisoning recognized in Thailand.

Lipophorin as a lipid carrier in the hemolymph of the horseshoe crab, Limulus polyphemus

A high-density lipoprotein (HDL) was obtained from the hemolymph of Limulus polyphemus in yields generally less than 30 micrograms/ml by ultracentrifugal flotation. SDS-PAGE revealed two apolipoproteins with masses similar to those of apolipophorins (apoLp-I, 265 +/- 14 kDa; apoLp-II, 89 +/- 6 kDa). Lipid composition was different from both insect lipophorin and crustacean HDL, and showed less diacylglycerols than triacylglycerols (3.8 and 36.2 of total lipids, respectively). Since Limulus polyphemus is closely related to precambrian chelicerates, our results confirm that lipophorin was present early in the evolution of arthropods.

C-reactive protein (CRP) in haemolymph of a mollusc, Achatina fulica Bowdich.

C-reactive protein (CRP) was found to be one of the major components of the haemolymph of A. fulica with a normal level of approximately 2 mg CRP/ml haemolymph. Immunological cross-reactivity was found between CRP from A. fulica, a mollusc and Limulus polyphemus, an arthropod. The present finding demonstrates the evolutionary significance of C-reactive protein.