Bacillus sphaericus binary toxin elicits host cell autophagy as a response to intoxication.

Bacillus sphaericus strains that produce the binary toxin (Bin) are highly toxic to Culex and Anopheles mosquitoes, and have been used since the late 1980s as a biopesticide for the control of these vectors of infectious disease agents. The Bin toxin produced by these strains targets mosquito larval midgut epithelial cells where it binds to Cpm1 (Culex pipiens maltase 1) a digestive enzyme, and causes severe intracellular damage, including a dramatic cytoplasmic vacuolation. The intoxication of mammalian epithelial MDCK cells engineered to express Cpm1 mimics the cytopathologies observed in mosquito enterocytes following Bin ingestion: pore formation and vacuolation. In this study we demonstrate that Bin-induced vacuolisation is a transient phenomenon that affects autolysosomes. In addition, we show that this vacuolisation is associated with induction of autophagy in intoxicated cells. Furthermore, we report that after internalization, Bin reaches the recycling endosomes but is not localized either within the vacuolating autolysosomes or within any other degradative compartment. Our observations reveal that Bin elicits autophagy as the cell's response to intoxication while protecting itself from degradation through trafficking towards the recycling pathways.

[A case of fatal neonatal Bacillus cereus meningitis]. / Méningite néonatale à Bacillus cereus, à propos d'un cas.

A fatal case of meningoencephalitis caused by Bacillus cereus, an uncommon but potential pathogen, resistant to most beta-lactam antibiotics, is described in a 28-day old premature neonate. Difficulties for clinical diagnosis and treatment are discussed. A review of the literature (26 published cases) is given. Early diagnosis of neonatal B. cereus infection is crucial as it leads to a standard treatment including vancomycin.

Acute inflammation and loss of retinal architecture and function during experimental Bacillus endophthalmitis.

Rapid vision loss and explosive inflammation are devastating consequences of Bacillus endophthalmitis that have not been well defined. We therefore analyzed the evolution of intraocular inflammation and loss of retinal architecture and function during experimental Bacillus endophthalmitis. Mice were intravitreally injected with 100 CFU of B. cereus, and eyes were analyzed for bacterial growth, retinal function, architectural changes and retinal cellular stress, inflammatory cytokines, and infiltrating cells. Retinal electrophysiologic and structural changes began as early as 4 to 6 hr postinfection. Significant declines in retinal function paralleled the loss of retinal architecture. Glial fibrillary acidic protein (GFAP) was detected in retina, indicating potential stress. Polymorphonuclear leukocyte (PMN) infiltration into the vitreous began as early as 4 hr postinfection, coinciding with a significant increase in TNF-alpha in the eye. These results indicated that acute inflammation and detrimental architectural and electrophysiologic changes in the retina began earlier than once thought, suggesting that therapeutic intervention should be given at the earliest possible time to avoid vision loss during Bacillus endophthalmitis.

Necrotizing gastritis due to Bacillus cereus in an immunocompromised patient.

Bacillus cereus is increasingly being acknowledged as a serious bacterial pathogen in immunocompromised patients. We present a case of acute necrotizing gastritis caused by B. cereus in a 37-year-old woman with acute myeloblastic leukemia, who recovered following total parenteral nutrition and treatment with imipenem and vancomycin. B. cereus was isolated from gastric mucosa and blood cultures. Up to now, no case of acute necrotizing gastritis due to this organism has been reported.

Bacillus endophthalmitis: roles of bacterial toxins and motility during infection.

PURPOSE: Bacillus endophthalmitis is a highly explosive infection of the eye that commonly results in rapid inflammation and vision loss, if not loss of the eye itself, within a few days. Quorum-sensing-controlled toxins are essential to virulence during infection. Another unique characteristic of this disease is the ability of Bacillus to replicate rapidly and migrate to all parts of the eye. This study was conducted to determine the combined roles of toxins and motility during Bacillus endophthalmitis. METHODS: Rabbit eyes were injected intravitreally with approximately 100 cfu of wild type, nonmotile, or nonmotile/quorum-sensing-deficient Bacillus thuringiensis. Infection courses were analyzed by biomicroscopy, histology, electroretinography, and quantitation of bacteria and inflammatory cells. RESULTS: Infection with wild type B. thuringiensis resulted in complete retinal function loss by 18 hours after infection, whereas nonmotile B. thuringiensis infections required 30 hours to achieve a reduction of >90% in retinal function. Further attenuation of infection resulted from infection with the nonmotile/quorum-sensing-deficient B. thuringiensis strain, with approximately 90% retinal function loss occurring at 36 hours. Overall, the nonmotile and nonmotile/quorum-sensing-deficient mutants were significantly less virulent than wild-type B. thuringiensis. CONCLUSIONS: The results demonstrate that, in addition to quorum-sensing-controlled toxin production, bacterial migration within the eye contributed to the rapidly fulminant and destructive course of Bacillus endophthalmitis. Motility and quorum-sensing may therefore represent possible targets for the development of therapies designed to attenuate the devastating effects of Bacillus in the eye during endophthalmitis.

Characterization of Mexican Bacillus thuringiensis strains toxic for lepidopteran and coleopteran larvae.

Bacillus thuringiensis strains C-4, C-9, GM-7, and GM-10, isolated from northeast Mexico and selected for their high toxicity against lepidopteran and coleopteran pests, were characterized following United States Environmental Protection Agency (EPA)'s guidelines. Flagellar serotyping revealed that GM-7 and GM-10 belonged to serotype aizawai, whereas C-4, C-9 corresponded to the kumamotoensis serotype. GM-10 and C-9 were also shown to be the most effective against lepidoptera and coleoptera larvae, respectively. None of the tested strains produced beta-exotoxin or showed activity against mosquitoes. GM-7 and GM-10 were sensitive to R-41 and CP-51 phages. All strains synthesized crystal proteins of 130-140 kDa. PCR analysis showed that C-4, GM-7, and GM-10 strains expressed cry1 genes, and C-9 expressed cry3 and cry7/8 genes, but not cry1. However, the C-9 strain had no cross-reaction with antisera raised against Cry3A and Cry7A proteins. GM-7 and GM-10 were sensitive to R-41 and CP-51 phages. When the delta-endotoxin (crystal) from the four strains was subcutaneously injected to Balb/c mice, alone or in combination with spores, only C-4 and C-9 provoked tissue necrosis similar to that caused by the beta-exotoxin producer HD-41. Tissue necrosis was prevented with the injection of pentoxifylline, an inhibitor of tumor necrosis factor alpha (TNF-alpha) production, suggesting a role of this cytokine in the observed effect. Our results demonstrated that GM-7 and GM-10 strains are effective and suitable for control of lepidopteran pests and safe for mammals under EPA regulations. The potential of the C-9 strain for the control of several coleopteran pests, and the induction of tissue necrosis in mice by C-4 and C-9 strains, are discussed.

[Pathogenetic mechanisms of bacterial cholangitis in mechanical jaundice (experimental histological study)]. / Patogeneticheskie mekhanizmy bakterial'nogo kholangita pri mekhanicheskoi zheltukhe (éksperimental'no-gistologicheskoe issledovanie).

In 89 rats with experimentally induced mechanical jaundice the infectious process was modeled by intragastric administration of E. coli and B. subtilis. The study included the investigation of the main routes and time intervals of dissemination, organ content of microbes labeled with 3H-thymidine, as well as of structural and functional peculiarities of the interaction of animal tissues with the microbes in cholestasis development dynamics and under the condition of biliary tract decompression. It was established that under the condition of mechanical jaundice viable bacteria from the alimentary tract penetrated the organs of peritoneal cavity through the blood and lymph flow. In the liver, the endogenous agents of surgical infections, and E. coli, in particular, were found to aggravate the inflammatory and necrobiotic processes. B. subtilis did not cause any inflammatory or other pathologic process in the tissue of liver.

Contribution of membrane-damaging toxins to Bacillus endophthalmitis pathogenesis.

Membrane-damaging toxins are thought to be responsible for the explosive clinical course of Bacillus endophthalmitis. This study analyzed the contribution of phosphatidylinositol-specific phospholipase C (PI-PLC) and phosphatidylcholine-specific phospholipase C (PC-PLC) to the pathogenesis of experimental Bacillus endophthalmitis. Isogenic mutants were constructed by insertion of lacZ into Bacillus thuringiensis genes encoding PI-PLC (plcA) and PC-PLC (plcB). Rabbit eyes were injected intravitreally with 2 log(10) CFU of strain BT407 (wild type), the PI-PLC mutant (BTplcA::lacZ), or the PC-PLC mutant (BTplcB::lacZ). The rates of decrease in retinal responses of eyes infected with the isogenic mutants were similar to that of wild type, with all infections resulting in elimination of retinal function by 18 h. Strain BT407 caused a significant increase in the latency of retinal responses at 6 h, but strains BTplcA::lacZ and BTplcB::lacZ did not. All strains elicited significant inflammatory cell influx into the anterior chamber by 12 h. Histologically, eyes infected with each strain were indistinguishable throughout the infection course. In this model, neither PI-PLC nor PC-PLC had an effect on the course or severity of experimental Bacillus endophthalmitis. Alterations in retinal responses early in infection may mark the beginnings of specific photoreceptor or glial cell dysfunction.

Effects of intravitreal corticosteroids in the treatment of Bacillus cereus endophthalmitis.

OBJECTIVE: To investigate whether intravitreal corticosteroid therapy reduces the extent of inflammatory intraocular tissue damage caused by Bacillus cereus endophthalmitis. METHODS: New Zealand white rabbits were inoculated with 1 x 10(6) B cereus organisms and randomized to receive no treatment (control eyes; n=14), intravitreal vancomycin hydrochloride (n=13), or a combination of intravitreal vancomycin and dexamethasone sodium phosphate (n=13) after 24 hours. The eyes were examined and graded for clinical signs of infection and inflammation on days 7 and 14, followed by enucleation for histopathologic analysis. RESULTS: Both treated groups had significantly less clinical sequelae than controls on day 7. By day 14, eyes given combination treatment had significantly less clinically graded corneal (P=.03) and conjunctival (P=.007) inflammation than eyes treated with vancomycin. Histopathologic analysis revealed a significant decrease in inflammatory changes between all treated eyes and controls at day 14. The only statistically significant difference between eyes given combination treatment and eyes given vancomycin alone was in the retina (P=.03). CONCLUSIONS: Intravitreal corticosteroids may enhance the recovery from B cereus endophthalmitis when given in conjunction with intravitreal antibiotics. The beneficial effect of corticosteroids is noted clinically, but not histologically, by day 14 after single-dose treatment in rabbits. CLINICAL RELEVANCE: This study provides evidence that the use of intravitreal corticosteroids with antibiotics for the treatment of B cereus endophthalmitis may lead to an improvement compared with the use of antibiotics alone. Arch Ophthalmol. 2000;118:803-806

Bacillus thuringiensis subsp. konkukian (serotype H34) superinfection: case report and experimental evidence of pathogenicity in immunosuppressed mice.

We present a case of severe war wounds infected by Bacillus thuringiensis serotype H34 and describe the experimental protocol used to demonstrate its ability to infect mice after cutaneous inoculation. This case is interesting because B. thuringiensis is considered to be a contaminant in laboratories and receives inadequate attention.

Histopathological and histochemical changes in honeybee larvae (Apis mellifera L.) after infection with Bacillus larvae, the causative agent of American foulbrood disease.

Morphological, histochemical and cytochemical changes were examined in honeybee larvae after infection with the bacterium Bacillus larvae. The results indicate cell necrosis in the midgut epithelium accompanied by increasing cell vacuolization and nuclear pyknosis following per os inoculation with B. larvae. Many autolysosomes were positive for acid phosphatase. Non-vacuolar acid phosphatase activity was also found in lysed cell compartments. No such activity was found in regenerative epithelial cells. Degradation of haemocytes, salivary glands and other tissues was also observed. Histochemical analyses after per cutaneous inoculation with B. larvae of three- and five-day-old honeybee larvae show intense non-vacuolar acid phosphatase activity followed by disintegration of infected salivary glands, epithelial cell cytoplasm and haemocytes.

Association of cilia-associated respiratory (CAR) bacillus with natural chronic tracheitis in goats.

A histological, histochemical and immunohistological study of the respiratory tract of 83 slaughtered goats (50 adults and 33 kids) is described. Cilia-associated respiratory (CAR) bacillus was detected by means of the Warthin Starry method in the tracheal epithelium of seven (21.2%) of the kids and 16 (32%) of the adult goats. A chronic diffuse tracheitis characterized by mixed lymphocyte and plasma-cell infiltration was found in all seven kids and in 17 adults, including the 16 infected with the CAR bacillus. Although not proved, it is possible that the CAR bacillus caused the chronic tracheitis. Immunohistochemical results suggested that the caprine CAR bacillus was closely related to the rabbit CAR bacillus.

Fluoroscopically guided percutaneous disk aspiration in 10 dogs with diskospondylitis.

Fluoroscopically guided percutaneous fine-needle aspiration of the intervertebral disk space was performed in 10 dogs with diskospondylitis. Positive bacterial cultures were obtained from 9 of 12 aspirated disk spaces, 1 of 6 blood cultures, and 6 of 10 urine cultures. Positive disk cultures were obtained from 2 dogs with negative blood and urine cultures and from 2 additional dogs with low numbers of Staphylococcus in urine cultures. Adverse clinical sequelae of the procedure were not noted. Percutaneous fine-needle aspiration of the intervertebral disk space is an alternative technique to surgical biopsy to obtain positive bacterial cultures from dogs with diskospondylitis.

Demonstration of rat CAR bacillus using a labelled streptavidin biotin (LSAB) method.

Immunohistochemical detection of rat CAR bacillus antigen in paraffin-embedded experimentally infected rat lungs, using an immunoperoxidase technique based on the labelled streptavidin biotin (LSAB) method and 3-amino-9-ethylcarbazole (AEC) as substrate is described in this paper. The pattern of immunostaining was confined to the ciliated bronchial epithelium and the specificity of this technique was confirmed. The use of AEC as substrate was evaluated more efficient than diaminobenzidine (DAB). The usefulness of this immunoperoxidase technique for the detection of CAR bacillus in rats and its advantages compared to the indirect immunofluorescence (IF) are discussed.

[The detection of Peptostreptococcus asaccharolyticus and Bacillus cereus in clinical materials from a child who died of sepsis with a fulminant clinical course]. / Vyiavlenie Peptostreptococcus asaccharolyticus i Bacillus cereus v klinicheskikh materialakh ot rebenka, umershego ot sepsisa s molnienosnym klinicheskim techeniem.

A case of an acute disease with a rapid clinical course and a fatal outcome in the presence of irreversible toxicoinfectious shock, appearing in two children after the consumption of sheep kidneys, is described. The post mortem examination of the children revealed the presence of hemorrhagic, erosive and necrotic areas on the mucous membrane of the stomach, the duodenum and the upper section of the small intestine. From the material obtained by probing the stomach of one of the children 6 hours before death P.asaccharolyticus and B.cereus were isolated. Hemorrhage, erosions and necrosis were also found in experimental mice, injected with the centrifugates of the gastric secretions of the patient and the cultures of the isolated bacteria, which was indicative of the presence of highly active exotoxin. On the basis of the above facts, compared with similar data in the literature, this case was considered to be etiologically related anaerobic Peptostreptococcus in symbiosis with B.cereus.

Experimental posttraumatic Bacillus cereus endophthalmitis in a swine model. Efficacy of intravitreal ciprofloxacin, vancomycin, and imipenem.

PURPOSE: The authors compare the intravitreal efficacy of ciprofloxacin, vancomycin and imipenem, in treating experimental Bacillus cereus endophthalmitis. METHODS: Thirty-three Yorkshire pigs received a surgically induced injury to the right eye, which was then repaired and injected with 8400 colony forming units of live B. cereus. Nine pigs received no therapy and served as a natural history group. Twenty-four other pigs then were randomized into a treatment group with ciprofloxacin (n = 6), vancomycin (n = 6), imipenem (n = 6), or normal saline (n = 6). Eyes were examined clinically 1, 2, 3, 4, 5, 6, 8, 12, and 24 hours after inoculation. After 24 hours, the eyes were enucleated for histologic study. RESULTS: Experimental disease was characterized by an aggressively developing endophthalmitis, with retinitis and vitritis developing at 4 hours. Histologic examination showed vitreous abscess and retinal necrosis. Both vancomycin- and imipenem-treated group had less inflammation and tissue destruction than control animals, based on the Wilcoxon rank sum test (P < 0.05). Ciprofloxacin-treated animals showed significantly more intraocular destruction and were indistinguishable from controls. CONCLUSION: Vancomycin and imipenem appear to limit inflammation and tissue destruction when given early in the course of experimental posttraumatic endophthalmitis caused by B. cereus. Results with ciprofloxacin are less conclusive and warrant further investigation.

[Intracerebral hemorrhage and multiple brain abscesses caused by Bacillus cereus within the scope of acute lymphatic leukemia]. / Intrazerebrale Blutung und multiple Hirnabszesse durch Bacillus cereus im Rahmen einer akuten lymphatischen Leukämie.

Multiple hematogenic brain abscesses in immunosuppression are occasionally caused by rare and primary apathogenic causative agents. We report a first case of an isolated CNS infection by Bacillus cereus, which led to death from multiple brain abscesses and an intracerebral hemorrhage, probably caused by the infection, within 4 days. The underlying disease leading to immunosuppression was acute lymphatic leukemia in complete remission. In spite of antibiotic therapy the chemotherapy-induced neutropenia enabled unhindered spreading of the necrotizing infection, which was verified by histological analysis. The production of potent toxins such as hemolysin and cerelolysin by B. cereus leads to rapid and fulminant tissue destruction usually involving the walls of blood vessels.

Fatal Bacillus cereus meningoencephalitis in an adult with acute myelogenous leukemia.

Bacillus cereus, a ubiquitous, endospore-forming, aerobic gram-positive bacillus, is primarily associated with toxin-mediated food poisoning. Frequently, isolates of Bacillus species from clinical specimens are discussed as contaminants. We report a rapidly fatal case of disseminated infection due to B cereus in a patient receiving induction chemotherapy for M0 acute leukemia. A short clinical syndrome of nausea and vomiting preceded neurologic symptoms. Autopsy showed extensive meningoencephalitis with subarachnoid hemorrhage and multiple liver abscesses. Areas of necrosis were devoid of any inflammatory response consistent with a severely immunocompromised state. The organism was isolated from immediate premortem and autopsy blood specimens. This case illustrates the possibility and severity of true B cereus infections in immunocompromised patients, the clinicopathologic features of which are as yet not well defined.