Mycobacterium avium and Mycobacterium intracellulare infections in patients with and without AIDS.

A genetic probe (Gen-Probe) was used to evaluate potential epidemiologic and susceptibility differences of Mycobacterium avium complex (MAC) strains isolated from 154 patients with and without the acquired immunodeficiency syndrome (AIDS). Genetic analysis revealed that 98% of the 45 patients with AIDS harbored only M. avium regardless of the anatomic or geographic source of the isolate; in contrast, approximately 40% of MAC isolates recovered from 109 patients without AIDS were M. intracellulare. Most M. intracellulare of respiratory origin recovered from patients without AIDS were involved in infectious processes. When 95 MAC isolates (M. avium, n = 53; M. intracellulare, n = 42) were evaluated for in vitro susceptibility to primary or secondary antimycobacterial drugs, significant differences were noted. M. intracellulare was more susceptible to streptomycin, rifampin, and ethambutol than M. avium; the converse was true for ethionamide. The results of this study suggest potentially important differences in disease spectrum and in vitro susceptibility profile for M. avium and M. intracellulare.

Quality control of individual components used in Middlebrook 7H10 medium for mycobacterial susceptibility testing.

The acceptability of different lots of commercial components which constitute our basal medium for susceptibility testing of mycobacteria was evaluated. The basal medium consisted of Middlebrook 7H10 agar supplemented with 10% oleic acid-albumin-dextrose-catalase and 0.5% glycerol. Studies were performed by using three separate microbiologic assays, and results were compared with parallel tests on previously standardized and acceptable lots of media. Components were rejected if comparison with standardized medium showed a major change in growth support or susceptibility status of any reference strain to any antimicrobial agent tested. Of the components tested in such a manner, 7 of 23 (30%) lots of 10% oleic acid-albumin-dextrose-catalase, 2 of 13 (15%) lots of Middlebrook 7H10 agar, and 0 of 5 lots of glycerol were found to be unacceptable. This study demonstrates that individual lots of components of this basal medium may vary significantly in their suitability for susceptibility testing, and failure to detect such variation may dramatically affect susceptibility profiles.

Improved selective medium for the isolation of lipase-positive Clostridium botulinum from feces of human infants.

Isolation of lipase-positive Clostridium botulinum from fecal specimens establishes the diagnosis of infant botulism, contributes to the diagnosis of food-borne botulism, and is most easily accomplished by use of selective media. Modification of an available selective medium, C. botulinum inhibitory medium (CBI), enabled more rapid isolation of C. botulinum. The modified medium (botulinum selective medium [BSM] contained (per liter) 25 g of dehydrated heart infusion broth, 20 g of agar, 30 ml of egg yolk suspension, 250 mg of cycloserine, 76 mg of sulfamethoxazole, 4 mg of trimethoprim, and 100 IU of thymidine phosphorylase at pH 7.4. The two media were compared by using 15 fresh fecal specimens from infant botulism patients (10 type A and 5 type B) and a C. botulinum isolate that had been obtained from an infant botulism patient and that was mixed into a fresh stool specimen from a healthy human infant. In comparison to CBI, BSM always provided better suppression of the nonbotulinum fecal flora and earlier emergence of lipase-positive colonies. Diagnosis of infant botulism was accomplished sooner with BSM than with CBI because isolation of lipase-positive C. botulinum was easier.

Protective role of human milk against sudden death from infant botulism.

We examined the possibility that human milk and formula milk might differentially affect the severity of the infant botulism because they differ in immunologic composition and in influence on the normal intestinal microflora against which Clostridium botulinum must compete. A beneficial effect of human milk was suggested by the different feeding experiences of the moderate, hospitalized patients and the sudden death cases. Of patients hospitalized in California, 66% (33/50) were still being nursed at onset of illness, a percentage significantly greater than that of matched controls (P less than 0.01). In contrast, all ten California cases of sudden infant death attributable to C. botulinum infection were being fed iron-supplemented formula milk at death) unlike their controls, P less than 0.02) and had received no human milk within ten weeks of death. A beneficial effect of human milk was also observed in differences in mean age at onset; hospitalized breast-fed patients were almost twice as old (13.8 +/- 6.7 weeks) as were hospitalized formula-fed patients (7.6 +/- 2.9 weeks) (P less than 0.01). Human milk (or possibly other factors associated with breast-feeding) appeared to have moderated the severity at onset of infant botulism, allowing time for hospital admission, whereas for some infants with this illness, formula milk (or possibly other factors associated with formula feeding) was linked to sudden unexpected death.

Effects of Clostridium botulinum toxins on infant and adult mice.

Toxic supernatants of many Clostridium botulinum type B isolates from various sources, especially those of isolates obtained from cases of infant botulism, appeared to more toxic in infant mice than in adult mice. The significance of this finding in diagnostic situations is discussed.

Quantitative evidence of intestinal colonization by Clostridium botulinum in four cases of infant botulism.

Infant botulism is an infectious form of a disease heretofore principally known as food-borne intoxication. Previous epidemiologic and laboratory studies have shown that infant botulism results from the ingestion of spores of Clostridium botulinum that subsequently germinate in the infant intestine and produce botulinal toxin. A quantitative study of the fecal microflora of four infants with infant botulism revealed the presence of C. botulinum in numbers as high as 6.0 x 10(8) colony-forming units (cfu)/g. At various times after the onset of illness, the numbers of C. botulinum that were recovered from feces ranged from 10(3) to 10(8) cfu/g and constituted from 0.01% to 3.3% of the total fecal flora. It was concluded that the large numbers of C. botulinum found in patients' feces could occur only as a consequence of in vivo spore germination and outgrowth.

Laboratory aspects of infant botulism in California.

Infant botulism is the newly recognized form of the disease in which illness results from the production of toxin in the infant's intestines. Between the recognition of infant botulism as a distinct clinical entity in 1976 and the end of 1978, 50 cases were identified in California. The diagnosis of botulism was established by the identification of botulinal toxin in and the isolation of Clostridium botulinum from stool specimens of the affected infants. Thirty cases were type A and 20 were type B. Fecal excretion of toxin and organisms was monitored during hospitalization and after discharge from the hospital. Stools from family members and from 160 age-matched control infants did not contain botulinal toxin. A total of 396 food and drug samples, as well as 159 environmental and miscellaneous items, were examined. None of the foods or drugs tested contained preformed toxin. However, C. botulinum organisms were isolated from the nine samples of honey, five of soil, and one of vacuum cleaner dust.

Food and environmental aspects of infant botulism in California.

In an effort to identify vehicles by which Clostridium botulinum spores might have reached the intestine of patients with infant botulism, 555 samples of foods, drugs, and environmental specimens were examined. Of the food items, C. botulinum was only found in nine of 90 (10%) honey specimens. Five patients had been exposed to honey that contained C. botulinum, and ingestion of honey was found to be a significant risk factor for type B infant botulism (P = 0.005). In addition, C. botulinum was isolated from five samples of soil (three from case homes, two from control homes) and from vacuum cleaner dust from one case home. In every instance in which C. botulinum was isolated from a specimen of honey, soil, or duct associated with a case of infant botulism, the type of toxin (A or B) in the honey, soil, or dust isolate matched the type of toxin of the organism recovered from the infant. Isolation of C. botulinum from the soil of homes of control infants emphasizes the ubiquitous distribution of and exposure to this organism and suggests that host factors are important in the development of illness. Prevention of infant botulism will depend on the identification of these host factors, as well as on the identification of other vehicles that, like honey, may convey C. botulinum spores to susceptible infants.

Honey and other environmental risk factors for infant botulism.

Infant botulism results from the in vivo production of toxin by Clostridium botulinum after it has colonized the infant's gut. Epidemiologic and laboratory investigations of this recently recognized disease were undertaken to identify risk factors and routes by which C. botulinum spores might reach susceptible infants. Clostridium botulinum organisms, but no preformed toxin, were identified in six different honey specimens fed to three California patients with infant botulism, as well as from 10% (9/90) of honey specimens studied. By food exposure history, honey was significantly associated with type B infant botulism (P = 0.005). In California, 29.2% (12/41) of hospitalized patients had been fed honey prior to onset of constipation; worldwide, honey exposure occurred in 34.7% (28/75) of hospitalized cases. Of all food items tested, only honey contained C. botulinum organisms. On household vacuum cleaner dust specimens and five soil specimens (three from case homes, two from control homes) contained Clostridium botulinum. The known ubiquitous distribution of C. botulinum implies that exposure to its spores is universal and that host factors contribute importantly to the pathogenesis of infant botulism. However, honey is now an identified and avoidable source of C. botulinum spores, and it therefore should not be fed to infants.

Isolation of Clostridium botulinum from Honey.

Methods for the isolation of Clostridium botulinum from honey samples are described. A total of 9 of 90 honey samples were positive for C. botulinum; 6 of the positive samples had been fed to babies who developed infant botulism.

Intestinal infection and toxin production by Clostridium botulinum as one cause of sudden infant death syndrome.

The spontaneous production of botulinum toxin in the infant gut by ingested Clostridium botulinum organisms is the underlying cause of infant botulism, recognised as an infectious disease only in late 1976. Because of the recognition of the pathophysiology of this disease and because the known potency and action of botulinum toxin can lead to rapid respiratory arrest, it appeared possible that the in-vivo production of botulinum toxin could cause the sudden death of some infants. To test this hypothesis, serum, selected tissues, and bowel contents from 280 dead infants were examined for the presence of C. botulinum toxin and/or organsisms. We found C. botulinum organisms in 10 infants, all of whom died suddenly and unexpectedly. 9 of these deaths were classified by the forensic pathologist as sudden infant death syndrome (S.I.D.S. or crib death). In 2 of these 10 sudden deaths both C. botulinum organisms and botulinum toxin were identified, and from the spleen of 1, C. botulinum organisms were isolated. Faecal specimens from 160 age-matched healthy infants who served as controls in studies of inpatient infant botulism cases were negative for both C. botulinum organisms and toxin, except for one specimen that contained only C. botulinum type A organisms. The 9 S.I.D.S. cases with evidence of C. botulinum infection comprised 4.3% of the 211 S.I.D.S. cases examined over 12 months. These findings suggest that intestinal production of botulinum toxin by C. botulinum is one cause of S.I.D.S. The strikingly similar age-distribution of 62 inpatient infant botulism cases and the 211 S.I.D.S. cases is also consistent with this concept. The possibility that in-vivo production of botulinum toxin may account for a larger proportion of S.I.D.S. cases is discussed.

Infant botulism. Epidemiological, clinical, and laboratory aspects.

Clostridium botulinum organisms and toxin were identified in the feces of six infants, aged 5 to 20 weeks, who had illnesses clinically consistent with botulism. Five of the infants lived in California and became ill within a six-month period in 1976; one infant became ill in New Jersey in 1975. Three cases were type A botulism, and three were type B. No source of ingested botulinal toxin could be found in any case. However, one infant with type B botulism had ingested a food containing C botulinum type B organisms, and no toxin was found in it. The clinical findings in these cases include constipation, weak sucking and crying ability, pooled oral secretions, cranial nerve deficits, generalized weakness, and, on occasion, sudden apnea. A characteristic electromyographic pattern termed "brief, small, abundant, motor-unit action potentials" (BSAP) was observed. The sources of C botulinum toxin for these six infants is thought to have been in vivo (gastrointestinal) production following ingestion of C botulinum organisms. Studies are underway to determine the full clinical spectrum, incidence, and potential public health importance of this infectious disease newly recognized in infants.

Infant botulism. Identification of Clostridium botulinum and its toxins in faeces.

Clostridium botulinum and its toxin were identified in the faeces of four infants, aged 6 to 13 weeks, who had symptoms consistent with botulism. Two cases had type-A toxin and two cases had type-B toxin present in their faeces. No toxin was detectable in sera C. botulinum and toxin could be recovered from faeces more than 8 weeks after admission to hospital. All four cases occurred within a 6-month period. The source of the toxin in these infants may have been in-vivo production from ingested organisms.

Clostridium botulinum type F: isolation from venison jerky.

A Clostridium botulinum type F was isolated from the venison jerky responsible for the only type F botulism outbreak reported in the United States. The isolate differed from the prototype Langeland type F strain in being nonproteolytic.