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Incidents of non-specific illness associated with the consumption of oysters have highlighted the lack of published information on the bacteriology of shellfish suitable for consumption. Investigations showed that the majority of molluscan shellfish entering English markets conform to the accepted standard of less than 5 Escherichia coli/ml. tissue. The numbers of E. coli were related to the sanitary quality of the growing area but no relation could be established between numbers of E. coli and coliforms, faecal streptococci or Clostridium welchii. The numbers of non-specific bacteria varied considerably but shellfish from sources associated with non-specific illness yielded relatively high counts at 37 degrees C. The results showed that there was no justification for a standard based on total plate counts, which often exceeded 10(6)/g. Such a standard would have to be coupled with spoilage or the incidence of non-specific illness. The relation between the numbers of non-specific bacteria growing at 20 and 37 degrees C. appears to be a useful measure for assessing the likelihood that raw shellfish are a public health risk.

The present study focused on the importance of contaminated sediments in shellfish accumulation of human viruses. Epifaunal (Crassostrea virginica) and infaunal (Mercenaria mercenaria) shellfish, placed on or in cores, were exposed to either resuspended or undisturbed sediments containing bound poliovirus type 1 (LSc 2ab). Consistent bioaccumulation by oysters (four of five trials) was only noted when sediment-bound viruses occurred in the water column. Virus accumulation was observed in a single instance where sediments remained in an undisturbed state. While the incidence of bioaccumulation was higher with resuspended rather than undisturbed contaminated sediment, the actual concentration of accumulated viruses was not significantly different. The accumulation of viruses from oysters residing on uninoculated sediments. When clams were exposed to undisturbed, virus-contaminated sediments, two of five shellfish pools yielded viral isolates. Bioaccumulation of undisturbed sediments by these bivalves was considered marginal when related to the concentration of virus in contaminated sediments; they would only represent a significant threat when suspended in the water column. Arguments were advanced for water-column sampling in the region of the water-sediment interface to provide an accurate determination of the virological quality of shellfish harvesting waters.

Consumption of raw shellfish has long been known to be associated with individual cases and sporadic outbreaks of enteric illness. However, during 1982, outbreaks of gastroenteritis associated with eating raw shellfish reached epidemic proportions in New York State. Between May 1 and December 31, there were 103 well-documented outbreaks in which 1017 persons became ill: 813 cases were related to eating clams, and 204 to eating oysters. The most common symptoms were diarrhea, nausea, abdominal cramps, and vomiting. Incubation periods were generally 24 to 48 hours long, and the duration of illness was 24 to 48 hours. Bacteriologic analyses of stool and shellfish specimens did not reveal a causative agent. Norwalk virus was implicated as the predominant etiologic agent by clinical features of the illness and by seroconversion and the formation of IgM antibody to Norwalk virus in paired serum samples from persons in five (71 percent) of seven outbreaks in which testing was done. In addition, Norwalk virus was identified by radioimmunoassay in clam and oyster specimens from two of the outbreaks. Determining the source of the shellfish was not always possible, but northeastern coastal waters were implicated. The magnitude, persistence, and widespread nature of these outbreaks raise further questions about the safety of consuming raw shellfish.

A new chromogenic compound, 5-bromo-4-chloro-3-indoxyl-beta-D-glucuronide, was found to be useful for the rapid, specific, differential identification of Escherichia coli in the sanitary analysis of shellfish and wastewater. Of 1,025 presumptively positive colonies (blue) and 583 presumptively negative colonies (nonblue), only 1% false-negative and 5% false-positive results were found.

The effects of storage on the microbial load in two commercially important species of shellfish were examined. Oysters (Crassostrea virginica) were stored as shellstock, shucked meats, and fully processed meats at four temperatures for up to 21 days, and clams (Mercenaria campechiensis) were stored only as shellstock. The concentrations of most microbiological groups of organisms increased with the duration and temperature of storage in both shellfish species, although the increases were significantly lower in claims. Concentrations of Vibrio cholerae rose by approximately 1 log in oysters stored as shellstock after 7 days at 2 degrees C, and Lac+ vibrios increased 2 logs at 8 degrees C. Total counts of bacteria, fungi, coliforms, fecal streptococci, Aeromonas hydrophila, and clostridia were significantly higher in shucked oysters than in those stored as shellstock. Fecal coliforms were statistically the same, but V. cholerae, Vibrio parahaemolyticus, and the Lac+ vibrios were higher in oysters stored as shellstock. The concentrations of all microbial groups were higher in fully processed oysters than in shucked meats, with the exception of the vibrios, which showed no significant difference among the treatments. The results showed that although traditional methods of storing shellfish resulted in an overall increase in the microbial load, vibrio levels increased only in oysters stored as shellstock. Although fecal coliform and total bacterial counts did not correlate with those for vibrios in fresh oysters, strong correlations were observed in oysters stored for 7 days, suggesting that these indicators may be useful in monitoring oyster quality when meats are stored for a limited time as shellstock.

Positive fecal coliform most-probable number results were compiled from 7,701 shellfish samples examined over 9 years. Generally, less than 2% of all positive reactions in EC medium are results of inoculations from presumptive media positive at the maximum (48-h) incubation period.

Vibrio vulnificus was enumerated in seawater and shellfish from two Florida estuaries at selected seasonal intervals. There were significant fluctuations in the presence and numbers of V. vulnificus. Relatively high seawater temperature and salinity favored the presence of V. vulnificus in both seawater and shellfish samples.

A study was carried out on the occurrence of Vibrio parahaemolyticus in Dutch mussels originating from the East Schelde Estuary. In a totol of 79 10-g tissue samples, 3 (3.8%) were found to contain V. parahaemolyticus. In a second survey, 6 out of 23 bags of mussels (26%) contained one or more strains of V. parahaemolyticus in 5-g tissue samples. The many limitations of current methodology used in such surveys are stressed. Positive samples can be missed because viable cells may die during refrigerated transport. Surviving cells also may not be detected because they have been sublethally stressed. In addition, the unreliability of the identification criterion of no growth in 10% NaCl was demonstrated.

A multiple-tube technique based on peptone water incubated at 44 degrees C for 24 h followed by detection of indole was found to be sensitive and specific for the detection of Escherichia coli in oysters and mussels. The method has the advantage of providing rapid results and is both less expensive and less time-consuming than other MPN techniques.

One hundred and ten hard clams (Mercenaria spp.) harvested from the Calabar river estuary were examined for total platable heterotrophic bacteria and Vibrio spp. in their mantle fluids using estuarine salts agar and thiosulphate citrate bile salts sucrose agar plates respectively. The mean counts of heterotrophic bacteria and vibrios were 7.9 x 10(6) organisms/ml and 2.5 x 10(2) vibrios/ml respectively. The rate of vibrio infection was 58.2%. Of the clams positive for vibrios, 54 (84%) harboured dual infections (V. alginolyticus and V. parahaemolyticus) of the moderate grade (greater than or equal to 10(3) to 10(5) vibrios/ml). Single infections due to V. alginolyticus or V. parahaemolyticus, found in 16% of clams, were either low (less than 10(3) vibrios/ml), moderate (greater than or equal to 10(3) to 10(5) vibrios/ml), or high (greater than 10(5) vibrios/ml). Dual infections only were detected in clams with mantle fluid volumes greater than 30 ml.

American oysters, Crassostrea virginica, and hard clams, Mercenaria mercenaria, were experimentally contaminated with Escherichia coli, Salmonella typhimurium, and Shigella flexneri either by intracardial injection or via the natural route of ingestion. Bacterial inactivation in the hemolymph was monitored for 72 h after exposure to these enteric pathogens at 20 and 6 degrees C. At 6 degrees C, both mean bacterial uptake by ingestion and subsequent clearance was singificantly lower that at 20 degrees C. However, substantial bacterial clearance from the hemolymph occurred for both shellfish at each temperature. At 20 degrees C, viable bacteria were no longer detectable after 24 h in hemolymph of either clams or oysters after exposure to contaminated water containing 4 x 10(3) bacteria per ml.

The most likely shellfish to be harvested if sportfishing is reinstated in San Francisco Bay is the Japanese cockle Tapes japonica. The virus levels present in these shellfish are unknown and need to be evaluated before the shellfish beds are open. Towards this end, a procedure for recovering and concentrating enteric viruses from these clams has been evaluated. Effective elution of poliovirus from clam tissues was found to occur with pH 9.5 glycine-buffered saline rather than with the pH 7.5 fluid utilized by other investigators on oysters. Poliovirus desorption was combined with Cat-Floc clarification to remove cytotoxicity from clam tissue homogenates. For assay purpose, viruses were concentrated by mixing the glycine supernatant with a beef extract solution, lowering the pH, and suspending the resulting floc in a small volume of phosphate buffer. This simple technique successfully recovered an average of 73% of the poliovirus added to clam homogenates at levels of 93 and 660 plaque-forming units per 100 g. Coxsackievirus B2 was isolated from clams exposed to raw sewage.

Samples of seawater, sediment, and the following invertebrates were collected from the Mediterranean Sea in the neighborhood of Alexandria, Egypt: sea urchins (Echinus spp.), clams (Tapes spp.), and wedge shells (Donax trunculus). A total of 165 samples were collected from November 1979 to July 1980. Average counts (per 100 mL or 100 g) of Vibrio parahaemolyticus content were as follows: seawater, 36; sea urchins, 349; sediment, 436; wedge shells, 534; clams, 1872. Samples collected in the summer months contained higher levels of V. parahaemolyticus than winter samples.

An outbreak of 17 cases of paralytic shellfish poisoning in humans occurred in Cape Town during May 1978. The clinical features were typical and no deaths occurred. Efforts to correlate the severity of disease with the amount of toxin ingested, to demonstrate a protective effect of alcohol, and to demonstrate the immunogenicity of the toxin proved unsuccessful. The regional ecological effects are described. Continued monitoring for the presence of toxic dinoflagellates must be conducted, and the dangers of the consumption of mussels from the Cape west coast should be widely publicized.

The incidence of bacteria of sanitary significance and isolation of Vibrio alginolyticus during a subtidal clam assessment study in the Bering Sea are described. Vibrio parahaemolyticus was not detected in clams, sediment, or seawater.

Vibrio parahaemolyticus was isolated from soft-shell clams (Mya arenaria) taken from 10 different clamming areas on Cape Cod, Mass., during July and August 1972. Direct plating on thiosulfate-citrate-bile salts-sucrose agar was found to be superior to either direct plating on Vanderzant modified salt starch agar or enrichment with Trypticase soy broth containing 7% salt for isolation from clam samples. Morphological and biochemical characteristics of 33 isolates from 30 samples generally conform to those described for this organism in the literature, except for the production of acid from sucrose, lactose, and sorbitol. Six of the isolates were hemolytic on human blood agar plates, whereas all showed a negative Kanagawa phenomenon. Twenty of the 33 isolates reacted with pooled antisera to the K antigen; 15 of these reacted with 9 different specific K antisera, leaving 5 untypable. Ten of these 15 reacted with 4 different O antisera.

Both biotypes of halophilous vibrios, V. parahaemolyticus and V. alginolyticus, have been found to cause intestinal diseases among the inhabitants of the littoral localities of the Crimea. These diseases mostly assume the form of acute gastroenteritis and alimentary toxic infections. Most frequently people contact infection by using sea food. It is suggested that the etiological unraveling of intestinal infections may be improved by introducing the method for the isolation of halophilous vibrios into laboratory practice.

The microbial cenosis in the mantle fluid and stomach of mussels is similar in its biochemical characteristics to the microflora of sea water.

Two strains of Vibrio parahaemolyticus were isolated from stool specimens of a patient who came down with gastroenteritis 20 hours after eating imported mussels which had not been cooked sufficiently. This is the first documented case of V. parahaemolyticus enteritis acquired in the Federal Republic of Germany.

The survival rate was followed up of S. typhimurium, S. enteritidis, S. aureus, and E. coli in the organs of M. galloprovincialis L. in experimental conditions. A vat was used with 30 mussels in 10 l of sea water contaminated with 10 cm3 broth culture in conc. of 2.10(4) bacterial cells per cub. cm. The water was changed at 24-hour intervals. Microbiologic studies were carried out of the gills, liver, and meat of the mussels and of the sampled sea water. Lowest survival rate showed S. aureus--up to the second day, S. typhimurium and S. orientalis--up to the 4th days (in the gills only), and E. coli--up to the sixth day (in the gills and liver). None of the experimental strains to the sixth day (in the gills and liver). None of the experimental strains was isolated from the meat. With regard to the periods of investigation cited M. galloprovincialis L. could play the role of a carrier, resp. vector of the infection and could bring about a disease after consumption. Stated is the important part played by mussels as biofilter, and, at the same time, as source of pathogenic bacterial microflora.

The accumulation of low levels (0.002 to 0.18 PFU/ml) of both feces-associated and monodispersed poliovirus by oysters (Crassostrea virginica or C. gigas) and clams (Mercenaria mercenaria) was investigated. These levels were chosen to duplicate the conditions present in light to moderately polluted waters. Experiments were performed in both small- and large-scale flowing seawater systems, developed to mimic the natural marine habitats of shellfish. Under these experimental conditions, viral accumulation by physiologically active shellfish was only noted when water column concentrations exceeded approximately 0.01 PFU/ml. Bioaccumulation increased with increasing concentrations of both monodispersed and feces-associated viruses. At virus concentrations below this level, viruses were seldom detected in either clams or oysters. Evidence indicated that the lack of accumulation was not the result of inefficient extraction or detection methods. The modified Cat-Floc-beef extract procedure used in the experiment was found to be capable of detecting as few as 1.5 to 2.0 PFU per shellfish. Evidence is presented to indicate that an uptake-depuration equilibrium was present at virus exposure levels of 0.10 PFU/ml, but not at 0.01 PFU/ml. The results suggested that viral accumulation by shellfish may not be efficient at water column concentrations below congruent to 0.01 PFU/ml.

The elimination of sewage effluent-associated poliovirus, Escherichia coli, and a 22-nm icosahedral coliphage by the common mussel, Mytilus edulis, was studied. Both laboratory-and commercial-scale recirculating, UV depuration systems were used in this study. In the laboratory system, the logarithms of the poliovirus, E. coli, and coliphage levels were reduced by 1.86, 2.9, and 2.16, respectively, within 52 h of depuration. The relative patterns and rates of elimination of the three organisms suggest that they are eliminated from mussels by different mechanisms during depuration under suitable conditions. Poliovirus was not included in experiments undertaken in the commercial-scale depuration system. The differences in the relative rates and patterns of elimination were maintained for E. coli and coliphage in this system, with the logarithm of the E. coli levels being reduced by 3.18 and the logarithm of the coliphage levels being reduced by 0.87. The results from both depuration systems suggest that E. coli is an inappropriate indicator of the efficiency of virus elimination during depuration. The coliphage used appears to be a more representative indicator. Depuration under stressful conditions appeared to have a negligible affect on poliovirus and coliphage elimination rates from mussels. However, the rate and pattern of E. coli elimination were dramatically affected by these conditions. Therefore, monitoring E. coli counts might prove useful in ensuring that mussels are functioning well during depuration.

This study is one of a series in which variations of the A-1 method for the detection and enumeration of fecal coliforms and Escherichia coli in seawater and foods were evaluated. The tests were conducted jointly by the Food and Drug Administration and state and provincial laboratories that support shellfish control programs in the United States and Canada as part of the National Shellfish Sanitation Program's Microbiology Task Force activities. Results showed significantly higher recovery of fecal coliforms from naturally contaminated shellfish by the AOAC official A-1-M method than by the American Public Health Association standard method. There was no significant difference in recovery of E. coli by the 2 methods.

Because the previous studies of 541 fish of the Southern Central Baltic Sea area gave negative results for the presence of Vibrio parahaemolyticus, 150 shellfish and 40 mud samples of the same origin were tested this time. In none of them presence of Vibrio parahaemolyticus nor Vibrio alginolyticus has been stated, though.

The bacterial flora of the Pacific oyster Crassostrea gigas, the sea mussel Perna viridis and the arkshell clam Scapharca cornea differed considerably from that of seawater in both numbers and generic composition. The numbers of heterotrophic bacteria in the bivalve shellfish, including the anaerobes and spore-forming bacteria, were greater than that in the surrounding water. Pseudomonas spp. were the dominant organisms, comprising over one third of the 321 strains characterized after isolation from the bivalves and seawater. Other bacteria isolated from the shellfish included Vibrio, Acinetobacter, and Aeromonas spp., whereas the seawater flora consisted mainly of coliform organisms, coryneform bacteria and Flavobacterium/Cytophaga spp. Bacteria associated with the deposit-feeding clams were higher in density and more distinct in generic composition as compared with those in the suspension-feeding oysters and mussels. Over 90% of the coliform and heterotrophic bacteria in oysters were found in organs associated with the digestive tract. Coliforms were mainly found in the stomach while heterotrophs were present in both stomach and the lower intestine. The results suggest that the stomach flora of oysters are mainly derived from the external environment and, through a process of selection and multiplication, that it may be gradually replaced by a more indigenous population which dominates the lower digestive tract.

Non-01 Vibrio cholerae gastroenteritis has been reported in the literature in association with foreign travel, or seafood ingestion, particularly raw shellfish harvested from warm-water climates. Non-01 V. cholerae gastroenteritis has not been reported before from New England. The case presented describes a previously healthy female who developed non-01 V. cholerae gastroenteritis approximately 26 hours after ingestion of raw cherrystone clams.

The relationship of fecal coliforms, Escherichia coli, and Salmonella spp. was examined in freshly harvested and stored shellfish. In 16 of 40 freshly collected oyster samples, fecal coliform levels were above the recommended wholesale level suggested by the National Shellfish Sanitation Program (less than or equal to 230/100 g), and Salmonella spp. were present in three of these samples. Salmonella spp. were not, however, present in any sample containing less than 230 fecal coliforms per 100 g. Analysis of the data suggests that low fecal coliform levels in both fresh and stored oysters are good indicators of the absence of Salmonella spp., but that high levels of fecal coliforms are somewhat limited in predicting the presence of Salmonella spp. E. coli levels correlated very strongly with fecal coliform levels in both fresh and stored oysters and clams, suggesting that there is no advantage in replacing fecal coliforms with E. coli as an indicator of shellfish quality.

Certain pathogenic micro-organisms in or on food of animal origin still constitute a particular hygienic risk. Salmonellae are found chiefly on meat and in meat products including poultry. Most of the infections with salmonellae in men are presumably caused by the consumption of raw products of animal origin. Our stock of slaughter animals is most often latently infected. During the slaughter and the processing meat surfaces are contaminated with salmonellae. Nevertheless a precautionary decontamination of these surfaces is not undertaken. Furthermore contaminations are still ignored by the official microbial meat inspection, and it is still allowed to use the meat for the production of minced meat meant to be eaten raw. Foods are also "infected" when the pathogenic organisms are introduced by contamination. It is not admissible to consider a small amount of salmonellae in food of animal origin as an acceptable lack of security that is to be respected by the consumer. Usual thinking models and customs are to be made dubious. It is shown at the example of raw meat and mussels that the hygiene of food must be optimized by comprising food technology.

Candida parapsilosis, C. tropicalis, and Torulopsis glabrata were the human-associated yeasts most frequently isolated from quahogs, oysters, and mussels collected from four estuarine areas along the northern shore of Long Island Sound. Some inconsistency and seasonal variation in the occurrence of these and other yeast species were noted. In particular, C. albicans densities were greatest during colder months in the more heavily polluted waters. A total of 347 yeasts were isolated and cultured at 37 degrees C and, of these, 219 of 62% were human-associated forms. Generally, these yeasts in the animals sampled reflected the overall pollution status of the estuary from which they were taken. This study represents a clear demonstration of potentially pathogenic yeasts in a valuable marine resource.

Codakia orbicularis may obtain nutrients from chemoautolithotrophic bacteria. The chemical composition of the C. orbicularis was investigated because of this unusual source of nutrition, and because it is a human food source in the Caribbean. The lipid fraction of these molluscs is discussed in detail. Polyunsaturated fatty acids account for only 11-15% of the total fatty acids, and non-methylene interrupted dienes are present as high as 9.5%. Cholesterol represents about 45% of the total sterols present.

The authors propose two new technics swift and accurate, to determine the sanitary quality of edible Bivalves. They experiment their technics on a sample of Mussels subjected in a conchal area to the method of self filtering.