Cases Of Botulism Research Articles

Use of enzyme-linked immunoassays for antibody to types C and D botulinum toxins for investigations of botulism in cattle.

The development of specific enzyme-linked immunosorbent assays (ELISA) for antibody to types C and D Clostridium botulinum toxins for investigation of botulism in cattle is described. Partially purified type C and D toxins were used as antigens to develop these ELISAs. Specificity of the ELISAs was evaluated on sera from 333 adult beef and dairy cattle from areas with no history or evidence of botulism in animals or water birds. The test was also evaluated on sera from 41 herds that included herds vaccinated against botulism, confirmed botulism cases and herds from areas where the disease is considered endemic. The ELISAs detected the presence of antibody to botulinum toxins in samples from vaccinated cattle and both convalescent and clinically normal animals from unvaccinated herds with outbreaks of botulism. Antibody was also found in unvaccinated animals from herds in which there had been no diagnosed botulism cases in areas where botulism was considered endemic. Sera from some unvaccinated cattle with high ELISA reactivity was shown to be protective for mice in botulinum toxin neutralisation tests. The use of these tests in investigations of botulism in cattle is discussed.

Wound Botulism—California, 1995

During January-November 1995, a total of 19 laboratory-confirmed cases of wound botulism were reported to the California Department of Health Services (CDHS); of these, 13 had occurred since August. Since 1990, the number of wound botulism cases reported annually in California has increased steadily (one case in 1990, two in 1991, three in 1992, four in 1993, and 11 in 1994). All cases except one since 1991 have occurred in injecting-drug users, and many involved subcutaneous injection or "skin popping" of black tar heroin. This report summarizes the findings of the investigation of two cases.

Foodborne Botulism—Oklahoma, 1994

On July 2, 1994, the Arkansas Department of Health and the Oklahoma State Department of Health were notified about a possible case of foodborne botulism. This report summarizes the investigation, which implicated consumption of improperly stored beef stew.

Survey of honey for Clostridium botulinum spores in eastern France

Ninety samples of honey produced in eastern France, and 26 samples of honey originating from various countries, were analyzed for the presence of Clostridium botulinum spores. Although eight samples were found positive when using the mouse toxicity test, none of the strains isolated from the positive samples were identified as C. botulinum. This result is consistent with the hitherto rare occurrence of cases of infant botulism reported in France.

Application of PCR to a clinical and environmental investigation of a case of equine botulism

PCR for the detection of botulinum neurotoxin gene types A to E was used in the investigation of a case of equine botulism. Samples from a foal diagnosed with toxicoinfectious botulism in 1985 were reanalyzed by PCR and the mouse bioassay in conjunction with an environmental survey. Neurotoxin B was detected by mouse bioassay in culture enrichments of serum, spleen, feces, and intestinal contents. PCR results compared well with mouse bioassay results, detecting type B neurotoxin genes in these samples and also in a liver sample. Other neurotoxin types were not detected by either test. Clostridium botulinum type B was shown to be prevalent in soils collected from the area in which the foal was raised. Four methods were used to test for the presence of botulinum neurotoxin-producing organisms in 66 soil samples taken within a 5-km radius: PCR and agarose gel electrophoresis (types A to E), PCR and an enzyme-linked assay (type B), hybridization of crude alkaline cell lysates with a type B-specific probe, and the mouse bioassay (all types). Fewer soil samples were positive for C. botulinum type B by the mouse bioassay (15%) than by any of the DNA-based detection systems. Hybridization of a type B-specific probe to DNA dot blots (26% of the samples were positive) and PCR-enzyme-linked assay (77% of the samples were positive) were used for the rapid analysis of large numbers of samples, with sensitivity limits of 3 x 10(6) and 3,000 cells, respectively. Conventional detection of PCR products by gel electrophoresis was the most sensitive method (300-cell limit), and in the present environmental survey, neurotoxin B genes only were detected in 94% of the samples.

Retrospective study of 108 cases of botulism in Poitiers, France.

Botulism, a food-borne toxin-mediated disease caused by Clostridium botulinum is still a common disease, which is most frequent in the rural environment; 108 cases, 66 males and 42 females, average age 32 years, were recorded from 1965 to 1990 in the infectious disease department of the University Hospital of Poitiers (France). In 83% of patients, the food responsible was home-cured ham. Mean incubation time was 3.4 days; digestive symptoms were observed in 93% of cases, ocular symptoms in 92% and urinary tract dysfunction in 22%. A scale of severity was used to classify the patients into those suffering from severe (6), intermediate (50) and mild (52) forms of the disease. Botulinum toxin type B was found in 36 (52%) of 69 blood samples and in 41 (51%) of 81 samples of the suspected food. From 1965 to 1976, 44 patients were treated with both toxoid and heterologous equine serotherapy. Since 1976, 29 patients have been treated with guanidine hydrochloride (35 mg/kg daily) and 35 patients with guanidine hydrochloride plus heterologous serotherapy. All 108 patients recovered without any sequelae.

A case of infant botulism associated with honey feeding in Italy

A case of infant botulism in a 9 week-old female is described. A strain of C. botulinum type B was isolated from the feces of the baby. The epidemiologic study detected in a sample of home canned honey Clostridium botulinum spores of the same serotype that was isolated from the patient. The honey had been used only to sweeten the pacifier of the baby. This is the first case of infant botulism in Europe linked conclusively to honey.

Transfer of neurotoxigenicity from Clostridium butyricum to a nontoxigenic Clostridium botulinum type E-like strain.

Two Clostridium butyricum strains from infant botulism cases produce a toxic molecule very similar to C. botulinum type E neurotoxin. Chromosomal, plasmid, and bacteriophage DNAs of toxigenic and nontoxigenic strains of C. butyricum and C. botulinum type E were probed with (i) a synthesized 30-mer oligonucleotide encoding part of the L chain of type E botulinum toxin and (ii) the DNA of phages lysogenizing these cultures. The toxin gene probe hybridized to the chromosomal DNA of toxigenic strains but not to their plasmid DNA. All toxigenic and most nontoxigenic strains tested were lysogenized by a prophage on the chromosome. Prophages of toxigenic strains, irrespective of species, had related or identical DNAs which differed from the DNAs of prophages in nontoxigenic strains. The prophage of toxigenic strains was adjacent or close to the toxin gene on the chromosome. Phage DNAs purified from toxigenic strains did not hybridize with the toxin gene probe but could act as the template of the polymerase chain reaction to amplify the toxin gene. The toxin gene was not transferred between C. botulinum and C. butyricum (either direction) when different pairs of a possible gene donor and a recipient strain were grown as mixed cultures. Nontoxigenic C. butyricum or C. botulinum type E-like strains did not become toxigenic when grown in broth containing the phage induced from a toxigenic strain of the other species.(ABSTRACT TRUNCATED AT 250 WORDS)

Infant botulism: A review in South Australia (1980–89)

Between March 1980 and March 1989 there were six identified cases of infant botulism. The presenting symptoms were predominantly poor feeding, lethargy and constipation, and the presenting signs were predominantly hypotonia, weak gag reflex and respiratory difficulty. All required intensive care and prolonged hospital stays. Recovery was eventually complete in all cases. Infant botulism is an uncommon condition but early recognition with adequate intensive care usually results in complete recovery.

A recent reminder of botulism

Botulism is a rare yet potentially common form of food poisoning that can be fatal. (1) Few documented cases of botulism exist in Australia (2,3,4) and New Zealand(5), emphasising how infrequently it is encountered. The recent admission to our Intensive Care Unit (ICU) of a man suspected of having botulism was a timely reminder of just how dangerous this condition can be. This paper will review the contemporary knowledge and interventions necessary in the management of botulism. It will also utilise anecdotes from our recent experience to illustrate some of the clinical scenarios and potentially fatal complications seen in this condition

Sequences of the botulinal neurotoxin E derived from Clostridium botulinum type E (strain beluga) and Clostridium butyricum (strains ATCC 43181 and ATCC 43755)

Recently, it has been shown that two Clostridium butyricum strains (ATCC 43181 and ATCC 43755), isolated from cases of infant botulism, produce a botulinal neurotoxin type E ( BoNT E ). Here we have determined the nucleotide sequences of the BoNT E genes of these two C. butyricum strains and from C. botulinum E strain Beluga. We show that the sequences of the BoNT E genes from the two C. butyricum strains are identical and differ in only 64 positions resulting in 39 amino acid changes (97% identity at the amino acid level) from that derived from C. botulinum. Our data suggest a transfer of the BoNT E gene from C. botulinum to the originally nontoxigenic C. butyricum strains.

Estimating the Value of Avoiding Morbidity and Mortality from Foodborne Illnesses

When people consume products violating the Food, Drug, and Cosmetic (FD&C) Act they may experience morbidity or mortality from foodborne illness. Some studies have used survey data to estimate the dollar value of avoiding a few illnesses, but surveys are expensive and the resulting estimates cannot easily be generalized to the wide variety of foodborne illnesses. We develop a method that uses published data to estimate the value of avoiding morbidity and mortality from foodborne illnesses using two metrics--quality-adjusted life-years and dollars. The method first describes the effects of different illnesses on the patient and then converts these descriptions into changes in time spent in different health states with different relative utility weights. We use these estimated changes to compute losses in quality-adjusted life-years. We demonstrate our method and derive estimates for the value of avoiding a future case of botulism, salmonellosis, chronic hepatitis, and bladder cancer. Researchers can use our method to compare the value of avoiding all illnesses caused by violations of the FD&C Act. More importantly, government officials can use our method to quickly and cost-effectively generate morbidity and mortality valuation estimates for any illness for policy purposes.

Infant Botulism with Down Syndrome

A 4-month-old boy with Down syndrome and infant botulism is reported. He was admitted to our hospital with stridor and developed respiratory failure and generalized paralysis. Laboratory examination revealed botulinal toxin type A in his blood and feces. Clostridium botulinum organisms were found in the feces and in the honey which he had been fed. Anti-botulinal toxin was administered on the 10th day of illness. Artificial ventilation was given for 87 days and he survived. This is the 11th known case of infant botulism in Japan. It is possible that other cases remain undiagnosed.

Therapeutic Uses of Botulinum Toxin

BOTULISM had been recognized by the 18th century, but the observation that a toxin produced by an anaerobic organism might be responsible for food poisoning was not made until 1897.1 Although seven immunologically distinct toxins have since been identified, only types A, B, and E have been linked to cases of botulism in humans.2 , 3 Botulinum toxin type A (hereafter referred to as botulinum toxin), one of the most lethal biologic toxins, has been found to be of therapeutic value in the treatment of a variety of neurologic and ophthalmologic disorders.4 The Food and Drug Administration recently approved botulinum toxin (Oculinum) . . .

Cloning of a DNA fragment encoding the 5'-terminus of the botulinum type E toxin gene from Clostridium butyricum strain BL6340

Chromosomal DNA was extracted from toxigenic Clostridium butyricum strain BL6340 isolated from a case of infant botulism. After digestion by EcoRI, a DNA fragment of about 1 kbp was cloned into Escherichia coli using lambda gt11, and was subcloned into pUC118. The E. coli cells transformed with this cloned fragment produced a 33 kDa protein which reacted with monoclonal antibodies recognizing the light chain (Lc) component of botulinum type E toxin. The nucleotide sequence of the cloned fragment was determined. The sequence was similar to that from botulinum type E toxin gene fragments previously determined by our laboratory (strains Mashike, Otaru and Iwanai). Several highly homologous sequences among the botulinum type A, C, E, butyricum and tetanus toxin genes were found in both translated and untranslated regions. These results suggest that the toxin gene of C. butyricum may have evolved by transfer from C. botulinum.

Effect of Heat and pH on Toxigenic Clostridium butyricum

ABSTRACT Clostridium butyricum long known to cause spoilage in canned acid foods recently caused two separate cases of infant botulism in Italy by strains producing type E botulinum toxin. Heat resistance of spores of toxigenic and nontoxigenic C. butyricum was determined. The non‐toxigenic strain was considerably more heat resistant, having a D‐value at 212°F of 4.7 min compared to a D‐value 170°F of 2.3–2.5 min for toxigenic strains at pH 7.0. Minimum pH for growth and toxin production was also determined. The nontoxigenic strain grew at pH 4.2; toxigenic strains grew and produced toxin at pH 5.2 but not at pH 5.0.

Garlic-in-oil associated botulism: episode leads to product modification.

In February 1989, three cases of botulism occurred in persons who consumed garlic bread made from a garlic-in-oil product. Testing of leftover garlic-in-oil showed it to have a pH of 5.7 and to contain high concentrations of Clostridium botulinum organisms and toxin. This was the second episode of botulism associated with a low acid garlic-in-oil product which needs constant refrigeration. In response, the Food and Drug Administration has taken steps to prevent a recurrence by requiring that microbial inhibitors or acidifying agents be added to such products.

ATTEMPTS TO QUANTITY CLOSTRIDIUM BOTULINUM TYPE A TOXIN AND ANTITOXIN IN SERUM OF TWO CASES OF INFANT BOTULISM IN JAPAN

Serum samples taken from two infant botulism cases during hospitalization were titrated for botulinum toxin by both the intraperitoneal (ip) injection method and the score method in mice. By the ip method, in which death is the only parameter, such low levels of toxin as lower than 4 ip LD50/ml may not be titrated even though the surviving mice show abdominal palsy. By the score method based on the degree of abdominal palsy, such low levels of toxin as 1.1 and 0.8 ip LD50/ml were detected in specimens of one of the patient's serum. No antitoxin was demonstrated in either case of infant botulism by applying the score method. It is not known whether spontaneous recovery from infant botulism is due to the antitoxin production.

COMPARATIVE STUDIES ON CLOSTRIDIUM BOTULINUM TYPE A STRAINS ASSOCIATED WITH INFANT BOTULISM IN JAPAN AND IN CALIFORNIA, USA

Twenty strains of Clostridium botulinum type A associated with infant botulism cases, six in Japan and 14 in California, USA, were compared in their characters. All six Japanese strains produced medium-sized progenitor toxin (M toxin; Mr 300 k) but no hemagglutinin and showed lower 50% infective doses (ID50) in the infant mouse test; whereas most American strains produced large-sized progenitor toxin (L + LL toxins; Mr 500 k and 900 k) and hemagglutinin in addition to M toxin and showed higher ID50 in infant mice. No marked difference in the biochemical properties was found between the two groups except for two American strains.

Risk Factors for Infant Botulism in the United States

To define risk factors for infant botulism, we performed a 2-year prospective case-control study of 68 laboratory-confirmed cases in infants living in the United States, outside of California. For each case patient, two control subjects were matched by date and hospital of birth or county birth records. By univariate analysis, breast-feeding (odds ratio = 2.9) and consumption of honey (odds ratio = 9.8) were associated with disease, but only 11 case patients (16%) had eaten honey. Decreased frequency of bowel movement (less than one per day for at least 2 months) was also associated with disease in infants 2 months of age and older (odds ratio = 5.2). Risk factors changed with the age of the patient at disease onset when analyzed by multivariate logistic regression methods. For infants less than 2 months old, living in a rural area or on a farm was the only significant risk factor (odds ratio = 6.4). For infants 2 months of age and older, breast-feeding (odds ratio = 3.8), less than one bowel movement per day for at least 2 months (odds ratio = 2.9), and ingestion of corn syrup (odds ratio = 5.2) were associated with disease. The severity of the disease was similar for breast- and bottle-fed infants. Clearly defined food exposures account for a minority of infant botulism cases. Preexisting host factors, such as intestinal flora and frequency of bowel movements, may be the most important risk factors for development of disease.