Showing papers on "Shigella dysenteriae published in 1995"

Genetics and regulation of heme iron transport in Shigella dysenteriae and detection of an analogous system in Escherichia coli O157:H7.

Abstract: Shigella species can use heme as the sole source of iron. In this work, the heme utilization locus of Shigella dysenteriae was cloned and characterized. A cosmid bank of S. dysenteriae serotype 1 DNA was constructed in an Escherichia coli siderophore synthesis mutant incapable of heme transport. A recombinant clone, pSHU12, carrying the heme utilization system of S. dysenteriae was isolated by screening on iron-poor medium supplemented with hemin. Transposon insertional mutagenesis and subcloning identified the region of DNA in pSHU12 responsible for the phenotype of heme utilization. Minicell analysis indicated that a 70-kDa protein encoded by this region was sufficient to allow heme utilization in E. coli. Synthesis of this protein, designated Shu (Shigella heme uptake), was induced by iron limitation. The 70-kDa protein is located in the outer membrane and binds heme, suggesting it is the S. dysenteriae heme receptor. Heme iron uptake was found to be TonB dependent in E. coli. Transformation of an E. coli hemA mutant with the heme utilization subclone, pSHU262, showed that heme could serve as a source of porphyrin as well as iron, indicating that the entire heme molecule is transported into the bacterial cell. DNA sequences homologous to shu were detected in strains of S. dysenteriae serotype 1 and E. coli O157:H7.

Persistence of local cytokine production in shigellosis in acute and convalescent stages.

Abstract: Shigella infection is accompanied by an intestinal activation of epithelial cells, T cells, and macrophages within the inflamed colonic mucosa. A prospective study was carried out to elucidate the cytokine pattern in Shigella infection linked to development of immunity and eradication of bacteria from the local site and also to correlate the cytokine profile with histological severity. An indirect immunohistochemical technique was used to determine the production and localization of various cytokines at the single-cell level in cryopreserved rectal biopsies from 24 patients with either Shigella dysenteriae type 1 (n = 18) or Shigella flexneri (n = 6) infection. The histopathological profile included presence of chronic inflammatory cells with or without neutrophils and microulcers in the lamina propria, crypt distortion, branching, and less frequently crypt abscesses. Patients had significantly higher (P 0.05) in the incidence of cytokine-producing cells between acute (2 to 8 days after the onset of diarrhea) and convalescent (30 days after onset) stages.

Immunoglobulin subclass distribution and dynamics of Shigella-specific antibody responses in serum and stool samples in shigellosis.

Abstract: To assess the humoral immunological responses at the subclass level in shigellosis, specific antibody responses against Shigella dysenteriae 1 lipopolysaccharide (LPS), S. flexneri Y LPS, invasion plasmid-coded protein antigens (Ipa), and Shiga toxin were analyzed. Antibody responses of 41 patients with S. dysenteriae 1 infection (SDIP) and 15 patients with S. flexneri infection (SFIP) were compared with those of controls (n = 40). The levels of total immunoglobulin G (IgG), IgA, IgM, and albumin in serum and stool samples were analyzed. In addition, total IgA (t-IgA), secretory IgA (s-IgA), and antigen-specific s-IgA in fecal samples were analyzed to evaluate the specificities and magnitudes of the mucosal immune responses. By comparing the relative increases in optical density for each IgG subclass separately, it was determined that the anti-LPS (homologous) response initially increased in the order IgG2 > IgG1 > IgG3 > IgG4 and that this order changed to IgG2 > IgG3 > IgG1 > IgG4 later in the disease. The IgG subclass response against protein antigens initially showed the order IgG1 > IgG3 > IgG2 > IgG4, which changed to IgG3 > IgG1 > IgG2 > IgG4 later in the disease. A significant increase in the proportion of IgA2 among t-IgA compared with that in controls was seen in both SDIP and SFIP, while significant changes in the proportions of IgG1 and IgG2 among t-IgG compared with controls was seen only in SDIP. The anti-LPS IgA2 response was more prominent in SDIP than in SFIP. We found an early peak of antigen-specific s-IgA in fecal samples, with a shorter duration than the corresponding response in serum samples. The simultaneous increase of serum IgA, fecal t-IgA, and s-IgA in SDIP compared with those in SFIP suggests that there is a massive increase in the local IgA production, giving an increase in systemic IgA concomitant with an extensive gut mucosal inflammation leading to an increased loss of albumin, IgG, and IgA with a high ratio of t-IgA to s-IgA.

Expression of flagella and motility by Shigella

Abstract: Since the discovery of Shigella as the aetiologic agent of acute dysentery almost 100 years ago, this organism has been described as a non-motile and nonflagellated organism that invades the human colonic mucosa. In this study, the production of flagella by prototypic strains of all four Shigella species and, moreover, by fresh clinical isolates was demonstrated by electron microscopy. The flagellum of Shigella (flash) is approximately 10 microns long and 12-14 nm in diameter and is typically seen emanating from one pole of the bacterium. Flash is composed of a putative structural polypeptide subunit of 33-38 kDa that shares immunological similarities with Escherichia coli, Salmonella spp., and Proteus mirabilis flagellins, and with the recently described recombinant Shigella flagellins (FliCSS and FliCSF) expressed in E. coli K-12. A fliCSS-specific oligo probe hybridized with all four Shigella species, while a fliCSF probe hybridized with all Shigella flexneri and Shigella dysenteriae strains, but not with all Shigella sonnei or Shigella boydii strains, indicating genetic divergence among their flagellin genes. Shigella exhibits motility in low-concentration motility agar under physiological growth conditions. The expression of flash and motility appears to be strictly regulated by unidentified genetic and environmental factors. These heretofore undescribed features may allow the bacteria to circumvent the natural intestinal mucosal defences leading to bacterial colonization and disease. The motility of shigellae may represent an evolutionary adaptation important for bacterial survival.

An animal model for the study of neurotoxicity of bacterial products and application of the model to demonstrate that Shiga toxin and lipopolysaccharide cooperate in inducing neurologic disorders

Abstract: An approach for studying neurotoxicity of bacterial products is presented. Pentylenetetrazol, a convulsant drug, was injected into mice, and increased sensitivity to pentylenetetrazol was used as an indicator of neurotoxicity. The preinjection of sonicates of Shigella dysenteriae 60R or Escherichia coli H30 (producing Shiga toxin or Shiga-like toxin I, respectively) enhanced the response of mice to pentylenetetrazol within 6 h. This was indicated by a higher mean convulsion score, increased number of mice responding with convulsions, and induction of seizures in animals pretreated with a subepileptic dose of pentylenetetrazol. Preinjection of purified Shiga toxin significantly changed the response to pentylenetetrazol only when coadministered with bacterial lipopolysaccharide (LPS); mean convulsion scores were 1.6 and 0.9 for the Shiga toxin-LPS group and controls, respectively. LPS alone did not affect sensitivity to pentylenetetrazol. These results suggest that Shiga toxin and LPS together induce neurologic disorders early in the course of infection.

Emergence of Nalidixic Acid Resistant Shigella dysenteriae type 1 in Thailand: An Outbreak Associated with Consumption of a Coconut Milk Dessert

Abstract: Shigella dysenteriae type 1 has been responsible for large outbreaks of severe dysentery in many parts of Asia but relatively few cases of the disease have been reported from Thailand and have generally not involved nalidixic acid resistant strains. Nalidixic acid resistant Shigella dysenteriae type 1 however emerged in a community in Thailand and was traced to a point source outbreak at a local school. Beginning March 1991 as part of prospective surveillance for diarrheal disease in Suan Phung all patients with diarrhea seen at the hospital outpatient department (OPD) in Suan Phung Thailand were cultured for enteric pathogens. 79 of the 197 patients treated for diarrhea at the hospital OPD in July and August 1992 had bloody diarrhea compared with 86/561 patients seen during 16 months of previous surveillance. Shigella dysenteriae type 1 was isolated from 33 of these 197 patients. Compared to matched controls patients with Shigella dysenteriae type 1 were more likely to attend one of the local elementary schools or live in the community surrounding that school. A cross-sectional study conducted at the school found that 50 of 485 students had dysentery in July. A coconut milk dessert prepared at the school was identified as the vehicle of transmission.

Structural elucidation of the O-antigen lipopolysaccharide from two strains of Plesiomonas shigelloides that share a type-specific antigen with Shigella flexneri 6, and the common group 1 antigen with Shigella flexneri spp and Shigella dysenteriae 1.

Abstract: Sugar and methylation analyses of native polysaccharides together with one-dimensional 1H- and 13C-NMR spectroscopy revealed that the two polysaccharides from strains 22074 and 12254 of Plesiomonas shigelloides are identical. The structure of the polysaccharide from strain 22074 was deduced from a uronic acid degradation and by NMR spectroscopy where heteronuclear multiple bond connectivitiy and two-dimensional nuclear Overhauser effect spectroscopy experiments established the pentasaccharide repeating unit as 4)-α-D-Galp A-(13)-α-D-Glcp NAc-(13)-α-L-Rhap -(12)-α-L-Rhap -(12)-α-L-Rhap -(1.

Epidemic dysentery caused by Shigella dysenteriae type 1: a sentinel site surveillance of antimicrobial resistance patterns in Burundi.

Abstract: Annual epidemics of bacillary dysentery have been a public health problem in Burundi for the last 14 years. Recent civil unrest, resulting in the displacement of large numbers of people into refugee settlements, has aggravated the situation. We report the results of a nationwide, health-centre based, sentinel site survey to check the drug resistance of Shigella dysenteriae type 1 (Sd1), the causal organism of such epidemics. Shigella spp. (of which 97% were Sd1) were isolated from 73% of the 126 specimens collected from six main sites around the country. There was no difference in culture results from fresh and frozen stool specimens. Overall Sd1 resistance to commonly available antibiotics (sulfamethoxazole + trimethoprim, ampicillin, tetracycline, and chloramphenicol) varied from 77% to 99% and was fairly uniformly distributed over the country. All Sd1 isolates were susceptible to newer drugs, such as ciprofloxacin and ceftriaxone. Resistance to nalidixic acid, the current first line of treatment for bacillary dysentery in Burundi, varied from 8% to 83% in the different sentinel sites; global resistance was 57%.

Detection of provisional serovars of Shigella dysenteriae and designation as S. dysenteriae serotypes 14 and 15.

Abstract: In initial studies of slide and tube agglutination tests of shigella-like isolates from diarrheal stools from Bangladesh, 10 and 8 strains of Shigella dysenteriae were identified as provisional serovars (serovars that have been defined but not given a number in the antigenic schema) E22383 and E23507, respectively. Further screening of diarrheal stool isolates in a slide agglutination test with antisera (made with the international reference strains) to the two provisional serovars of S. dysenteriae identified an additional 36 isolates as belonging to E22383 and 8 isolates as belonging to E23507. All strains had properties typical of invasive Shigella strains in that they were positive in the Sereny test and had the 120- to 140-MDa plasmid associated with invasiveness. On the basis of these markers of pathogenicity and isolation from various geographical locations, now we recommend that provisional serovars E22383 and E23507 be designated S. dysenteriae serotypes 14 and 15, respectively.

C reactive protein and prealbumin as markers of disease activity in shigellosis.

Abstract: To evaluate serum C reactive protein (CRP) and prealbumin concentrations as markers of disease activity in shigellosis this study serially measured serum concentrations of CRP and prealbumin in 39 patients infected with Shigella spp, and a comparison group of 10 patients infected with Vibrio cholerae serotype 01. On admission, patients with shigellosis had significantly higher median concentrations of CRP (109 v 5 mg/l; p or = 110 mg/l had a 70% sensitivity and a 61% specificity in predicting failure of treatment among patients infected with Shigella spp; the predictive value of a positive and negative test was 14% and 96% respectively. In summary, acute shigellosis elicits an acute phase response, the magnitude of which predicts clinical outcome.

Does treatment of bloody diarrhea due to Shigella dysenteriae type 1 with ampicillin precipitate hemolytic uremic syndrome

Abstract: Diarrhea-associated hemolytic uremic syndrome (HUS), the most common cause of acute renal failure in infancy and childhood, is often associated with infection by organisms producing Shiga toxin (ST) or Shiga-like toxin (SLT), mainly verocytotoxin-producing Escherichia coli (VTEC O157:H7) and Shigella dysenteriae type 1 (1,2). Although antibiotics are believed to be essential in treating shigellosis, treatment of S. dysenteriae type 1 patients with antibiotics to which the organism is resistant has been considered a risk factor for HUS (3,4). Until 1993, HUS was rarely reported from Saudi Arabia. Four cases of diarrhea-associated HUS due to S. dysenteriae type 1 were identified in 1989 (J. Hibbs and A. Mishkas, unpublished report), and one case of HUS attributed to plasma transfusion was documented in 1988 (5). In May 1993, four dysentery-associated HUS cases in two families were reported from northwestern Saudi Arabia (Tabuk). S. dysenteriae type 1 was isolated from the stool of each HUS patient. The organism was also isolated from 6 of the other 10 members of the two families who had dysentery. All isolates were resistant to trimethoprim-sulfamethoxazole, chloramphenicol, tetracycline, and ampicillin but sensitive to nalidixic acid. The two families had just returned from a 1-week visit to relatives in two neighboring villages in Gizan. This densely populated region in southwestern Saudi Arabia has about 1.2 million people living in more than 4,000 villages; the population is relatively poor and uneducated, and environmental sanitation is generally inadequate. We defined a case of HUS as any case of bloody diarrhea (BD) that had all of the following: acute renal failure (serum urea nitrogen, 18 mg/dL (or 6.3 mmol/L); or creatinine, 1.3 mg/dL (or 115 mmol/L)); thrombocytopenia (platelet count 130,000/mm3); and hemolytic anemia (hemoglobin level less than 10 g/dL; or hematocrit less than 30%; or appearance of fragmented red cells on direct microscopy). We standardized the treatment of BD in the Gizan region as follows: No antibiotics were given for treatment of BD at the primary health care centers (PHCCs) before a stool specimen was taken for culture and sensitivity testing. After reviewing the preliminary results, we either recommended use of nalidixic acid for treatment of BD or were guided by the results of the stool culture. This protocol was followed for management of BD in the entire region. Parasitologic, bacteriologic, and biochemical tests and drug treatment regimens were obtained for all patients admitted with BD or HUS to the regional referral hospital or five district hospitals in the outbreak area. BD cases were identified through hospital admission records, visits to PHCCs in the affected villages, interviews with family members of the identified patients, and school visits. We visited the houses of all HUS and BD patients and interviewed family members to ascertain which antibiotic was used to treat the BD patients; mothers were shown bottles and boxes of antibiotics and were asked to identify the antibiotic used for treating the children with BD. We identified 233 cases of BD occurring from February through July 1993 among 79 families scattered over 19 contiguous villages. Affected villages were predominantly in southern Gizan region near the Yemeni border. One hundred ninety patients (81.5%) consulted PHCCs; of those, 97 (51%) were referred to hospitals, and 81 (43%) were admitted. Thirty-four other BD patients were admitted directly to hospitals (a total of 115 admissions). In nine BD cases patients did not seek medical care including seven (3%) who used traditional treatment (the Wicka plant). In 23 (10%) patients, 13 male and 10 female, BD developed into HUS. Four isolates of S. dysenteriae type 1 that showed the same antibiotic susceptibility described earlier were obtained from four patients with BD in different villages in the middle of the outbreak. We used Cary-Blair transport medium for transporting stool specimens collected before antibiotic treatment from newly recognized patients with BD. However, communityand hospital-based interviews showed that the sequence of symptoms was almost identical in all of the 233 BD cases: the condition started with colicky abdominal pain and tenesmus (69%), followed by watery diarrhea (60%), which rapidly became only Dispatches

Serogroup prevalence of Shigellae in Bombay.

Abstract: Prevalence of Shigellae serotypes in Bombay was studied from June 1988 to May 1991. A total of 2758 faecal specimens were collected from paediatric patients (< 12 yrs) with acute gastroenteritis. A total of 90 Shigella were isolated giving the isolation rate of 3.2%. Shigella flexneri was the predominant serogroup (73.3%) followed by Shigella dysenteriae (16.6%). All the isolates were sensitive to nalidixic acid. Eighty percent of the Shigellae were multidrug resistant. Present data were compared with the study carried out during the period of 1983-87 from the same institute. A change in the serogroup prevalence was noted wherein Shigella flexneri dominated over Shigella dysenteriae since 1985. Increase in resistance to ampicillin and cotrimoxazole was seen in Shigella flexneri strains as compared to previous years.

Expression of Shigella dysenteriae serotype 1 O-antigenic polysaccharide by Shigella flexneri aroD vaccine candidates and different S. flexneri serotypes.

Abstract: The potential utility of Shigella flexneri aroD vaccine candidates for the development of bi- or multivalent vaccines has been explored by the introduction of the genetic determinants rfp and rfb for heterologous O antigen polysaccharide from Shigella dysenteriae serotype 1. The serotype Y vaccine strain SFL124 expressed the heterologous antigen qualitatively and quantitatively well, qualitatively in the sense of the O antigen polysaccharide being correctly linked to the S. flexneri lipopolysaccharide R3 core oligosaccharide and quantitatively in the sense that typical yields were obtained, with ratios of homologous to heterologous O antigen being 4:1 for one construct and 1:1 for another. Moreover, both polysaccharide chains were shown to be linked to position O-4 of the subterminal D-glucose residue of the R3 core. In contrast to the hybrid serotype Y SFL124 derivatives, analogous derivatives of serotype 2a vaccine strain SFL1070 did not elaborate a complete heterologous O antigen. Such derivatives, and analogous derivatives of rough, O antigen-negative mutants of SFL1070, formed instead a hybrid lipopolysaccharide molecule consisting of the S. flexneri lipid A R3 core with a single repeat unit of the S. dysenteriae type 1 O antigen. Introduction of the determinants for the S. dysenteriae type 1 O antigen into a second serotype 2a strain and into strains representing other serotypes of S. flexneri, revealed the following for the expression of the heterologous O antigen: serotypes 1a, 1b, 2a, and 5a did not produce the heterologous O antigen, whereas serotypes 2b, 3a, 3b, 4a, 4b, 5b, and X did.

Significance of the secreted form of IpaC, a 45 kDa protein of Shigella dysenteriae 1, in the invasive process as determined by monoclonal antibodies

Abstract: Invasion plasmid antigen C (IpaC), a 45 kDa plasmid encoded protein, is associated with the virulence of virulent Shigella spp. In S. dysenteriae type 1 the 45 kDa IpaC protein is secreted to a greater extent into the surrounding medium in comparison to other Shigella spp. Monoclonal antibodies (mAbs) to the secreted form of IpaC protein were raised in this study. Of the four secretory hybrid cells, one (3G4) was found to have a very high antibody titre as determined by ELISA. The specificity of 3G4 was confirmed by immunoblotting of whole cell extract of Escherichia coli strain MC1061 carrying the plasmid pHW756 which synthesizes both the IpaB and C proteins. The effect of the mAbs on plaque formation by virulent Shigella dysenteriae 1 was determined and it was found that the clone 3G4 substantially (55%) reduced plaque formation on HeLa cell monolayer. The epitope specificity of the mAb 3G4 was competitively inhibited by the convalescent phase sera from human, suggesting that the epitope recognized by clone 3G4 was expressed during the natural course of infection and also indicating that the 45 kDa (IpaC) protein in secreted form has a definite role in the invasive process.

Role of enteric nervous system in Shigella dysenteriae type 1 toxin-induced fluid secretion in rabbit ileum.

Abstract: Intracellular-free calcium concentrations were measured in the intestinal epithelial cells isolated from the rabbit ileum treated with Shigella dysenteriae type 1 toxin. There is almost a five-fold increase in the [Ca+2]i levels in the epithelial cells isolated from the toxin-treated rabbit ileum compared to control. The [Ca+2]i levels were also measured in the presence or absence of tetrodotoxin, a neurotoxin, and atropine, a muscarinic cholinergic antagonist. These drugs significantly decreased the [Ca+2]i levels of the cells isolated from the toxin-treated rabbit ileum. Furthermore, the unidirectional fluxes of Na+ and Cl- were determined in both the S. dysenteriae type 1 toxin-treated and control rabbit ileum. Dantrolene significantly stimulated net absorption of these ions in the toxin-treated ileum, suggesting the involvement of [Ca+2]i in the regulation of intestinal electrolyte transport. To evaluate the role of neural mechanisms, the transport of these ions was also determined in the presence and absence of tetrodotoxin and atropine. The presence of these drugs caused significant absorption of Na+ and Cl- in the toxin-treated compared to their absence. These findings suggest that the enteric nervous system plays an important role in S. dysenteriae type 1 toxin-induced diarrhoea, and the release of neurotransmitters causes changes in the intracellular calcium levels, ultimately changing electrolyte transport across the rabbit ileum.

[Hemolytic-uremic syndrome after verotoxin-producing Escherichia coli infection].

Abstract: The haemolytic uraemic syndrome, first described in 1955 by Gasser, is the number one cause of acute renal failure in infants. There are three types of the haemolytic uraemic syndrome: the seasonal epidemic form with prodromic diarrhoea and generally favourable outcome which usually occurs in infants, a less typical form without signs of digestive tract involvement and no seasonal prevalence which occurs more readily in older children and sometimes in families has a less favourable prognosis, and finally drug- or disease-related forms. Currently, overall mortality due to haemolytic uraemic syndrome has been reduced to about 4%, usually as a result of damage to the central nervous system. Several microorganism, including Shigella dysenteriae, Salmonella typhi, Campylobacter jejuni, Streptococcus pneumoniae, Rickittsiae and certain viruses (Coksackiae, Influenzae, Epstein-Barr) have been identified as causative agents. In 1983, digestive tract infection due to an Escherichia coli strain producing verotoxin was identified as capable of producing haemolytic uraemic syndrome and more rarely thrombopenic thrombotic purpura. The germ produces two exotoxins (whose effect is accentuated by the E. coli lipopolysaccharide endotoxin) which lead to the glomerular microangiopathy causing haemolytic uraemic syndrome. Diagnosis is based on identification (monoclonal antibodies, ELISA, PCR) of the verotoxins themselves or the two encoding genes in stool samples. Symptomatic treatment is essential but the effectiveness of antibiotics is still debated. Theoretically, antibiotics could worsen the syndrome by increasing endotoxin release from lysed bacteria, but inversely they could also prevent the syndrome if given early enough. Further research is required to acquire precise epidemiological data and identify animal reservoirs of verotoxin producing E. coli.

Antibiotic sensitivity to epidemic strains of Vibrio cholerae and Shigella dysenteriae 1 isolated in Rwandan refugee camps in Zaire

Abstract: Multiresistance or epidemic enteric bacteria to antibiotics greatly complicates treatment, and in some cases prophylaxis, of severe invasive gastroenteritis. During the summer of 1994, two epidemics of diarrhea, one due to Vibrio cholerae and the other to Shigella dysenteriae 1 isolated from the Goma and Bukavu camps was determined by measurement of the Agar Minimal Inhibitory Concentration. Multiresistance to tetracyclins, aminopenicillins, trimethoprimsulfamethoxazole, and nifuroxazide was observed. After intensive treatment mutant forms of both bacteria resistant to nalidixic acid rapidly appeared. Only fluoroquinolones remained active on these mutant strains, but the availability of this agent in Africa is restricted due to cost. The most effective way of preventing resistance is to limit the spread of enteric infections by health education and improvement of hygiene. This can be difficult during wartime.

Cytokines in the stools of children with complicated shigellosis.

Abstract: The pathogenesis of the systemic complications, leukemoid reaction and hemolytic uremic syndrome, associated with Shigella dysenteriae type 1 infection is not well understood. The excessive production of proinflammatory cytokines, including tumor necrosis factor alpha (TNF-alpha) and interleukin 6 (IL-6), has been suggested as a possible factor. We measured IL-6 and TNF-alpha in stools of 56 children with S. dysenteriae 1 infection and 29 children without any apparent infection, all age 12 to 60 months. Sixteen children with S. dysenteriae 1 infection had leukemoid reaction or hemolytic uremic syndrome (complicated shigellosis), while the others did not (uncomplicated shigellosis). Stool IL-6 and TNF-alpha concentrations were higher in children with uncomplicated shigellosis than in children with complicated shigellosis (P = 0.009 and < 0.001, respectively) or in uninfected children (P < 0.001). It is concluded that complicated infection is not associated with higher concentrations of the proinflammatory cytokines IL-6 and TNF-alpha in stool.

Shigellosis in Thai children: experience from a rural hospital 1985-1993

Abstract: Six hundred and ninety-four cases of shigellosis in Nakhon Nayok Hospital from January 1985 to December 1993 were studied to determine epidemiologic and microbiological features. Forty-five percent of cases were children under the age of 14 years. The majority of cases were in children under the age of four. The organism was found throughout the year, with peak incidence in June and July. The most common type isolated was Shigella flexneri, about 74.43%. Only 0.32% of organisms were Shigella dysenteriae. Shigella isolates showed a high rate of resistance to ampicillin and co-trimoxazole, in 1993 only 16.67% and 22.22% were sensitive respectively to these 2 drugs, but 100% were still sensitive to nalidixic acid. Fewer cases of shigellosis were isolated in recent years possible due to widespread use of quinolones in the treatment of acute infective diarrhea in adults.

Antimicrobial susceptibilities of Shigella dysenteriae type 1 isolated in Zimbabwe--implications for the management of dysentery.

Abstract: Shigella dysenteriae type 1 was cultured from 56/170 (33 pc) rectal swab specimens collected from patients presenting to hospitals in Harare, Zimbabwe with dysentery. All of the isolates were resistant in vitro to trimethoprim-sulfamethoxazole, with MICs > 32 mg/l, and all except one were resistant to ampicillin, most with an MIC > 256 mg/l. One isolate was resistant to nalidixic acid (MIC > 256 mg/l), but all of the others were sensitive, most with an MIC of 2 mg/l or less. Using antibiotic disks, 96 pc isolates were resistant to chloromphenicol and 94 pc to tetracycline. All isolates were sensitive in vitro to gentamicin. On the basis of these findings, we suggest that commonly available antibiotics including ampicillin, cotrimoxazole, chloramphenicol or tetracycline should not be used for the treatment of dysentery. The most appropriate antimicrobial agent at the present time would be nalidixic acid. Resistance to this is, however, likely to emerge and data on susceptibilities to fluoroquinolones as well as to cephalosporins should be obtained so that further recommendations can be given timeously.

Surface components of shigellae involved in adhesion and haemagglutination

Abstract: M.A. HAQUE, F. QADRI, K. OHKI AND O. KOHASHI. 1995. Strains of Shigella species were Studied for their ability to adhere and agglutinate mammalian erythrocytes. Shigella dysenteriae and Sh. flexneri exhibited haemagglutinating (HA) properties when cultured in Casamino Acids-Yeast Extract (CYE) broth in the presence of 1 mmol l -1 calcium chloride, but other shigellae did not show this property under the same culture conditions. Repeated subcultivation of Sh. boydii, Sh. sonnei and HA negative strains of Sh. dysenteriae and Sh. flexneri in CYE broth medium induced adhesive and haemagglutinating properties that were inhibited by sodium periodate. HA activities of Shigella spp. were also inhibited by N-acetylneuraminic acid, α 1 -glycoprotein and fetuin, but not by protease. Electron microscopy of Sh. dysenteriae 1, Sh.flexneri 2a, Sh. boydii 12 and Sh. sonnei 1 grown in CYE broth showed the presence of an extracellular slime layer that promoted agglutination of erythrocytes. The slime layer extracted from the cell surface of Shigella spp. showed HA properties, whereas lipopolysaccharide (LPS) obtained from the same strains, except Sh. dysenteriae 1, did not agglutinate erythrocytes. This evidence suggests that the cell surface haemagglutinin is a loosely bound slime layer which is expressed in CYE broth medium.

Fecal excretion of leukotriene C4 during human disease due to Shigella dysenteriae.

Abstract: Fecal excretion of leukotriene C 4 was determined in 26 individuals with dysentery and in 19 healthy controls. Of the patients, five were infected with Shigella dysenteriae type 1, 15 were injected with Shigella flexneri, two were infected with Shigella boydii, and hour were injected with Shigella sonnei. Three of the healthy controls were infected with non-dysenteriae Shigellae. All isolates of Shigella dysenteriae type I produced Shiga toxin; the other strains were not toxigenic. Patients with dysentery due to Shigella dysenteriae type I excreted higher concentrations of leukotriene C 4 (median, 3,234 pg/0.05 g of feces) than either ill individuals infected with non-dysenteriae Shigellae (median, 202 pg/0.05 g) or healthy criers (median, 145 pg/0.05 g) and uninfected controls (median, 129 pg/0.05 g). We propose that Shiga toxin stimulates intestinal mast cells, which release leukotriene C 4 , contributing to the inflammatory response in Shigella dysenteriae type I-associated dysentery

Peripheral blood neutrophil responses in children with shigellosis.

Abstract: Alterations in peripheral blood neutrophil function are known to occur in patients with colitis and may have a role in precipitating nonspecific tissue injury. It is not known whether neutrophil function is altered in patients with Shigella dysenteriae type 1 infection, during which there is extensive colitis and which may be associated with life-threatening complications in young children. Three aspects of peripheral blood neutrophil function, polarization, attachment to yeast particles, and locomotion, were therefore studied in 111 children with S. dysenteriae type 1 infection and 57 children without any infection. All children were aged 12 to 60 months. Of the children with S. dysenteriae type 1 infection, 42 had leukemoid reaction, hemolytic-uremic syndrome, or septicemia (complicated shigellosis), while the others did not (uncomplicated shigellosis). Polarization and locomotion in the absence of chemoattractants and in response to N-formylmethionyl-leucylphenylalanine (FMLP) and the lipopolysaccharide (LPS) of S. dysenteriae type 1 were determined. Attachment to unopsonized and opsonized yeast particles was also determined. Children with shigellosis (uncomplicated or complicated) had more polarized neutrophils with and without chemoattractants than uninfected children (P < 0.05). Children with complicated shigellosis had more polarized neutrophils with FMLP at 10(-7) and 10(-6) M (P < 0.05) and with LPS than children with uncomplicated shigellosis (P < 0.05). At 3 to 5 days after enrollment, the numbers of polarized neutrophils with 10(-8), 10(-6), and 10(-5) M FMLP declined in children with uncomplicated shigellosis but not in those with complicated shigellosis. Attachment to yeast particles was similar in all three groups of children. Locomotion was inhibited by LPS in children with shigellosis (P < 0.05), whether it was uncomplicated or complicated, compared with locomotion in uninfected children. Finally, neutrophil polarization in uninfected children was negatively influenced by nutritional status. Thus, poorly nourished uninfected children had more polarized neutrophils with FMLP at 10(-9) M (P < = 0.02) and 10(-5) M (P = 0.043) than their better-nourished counterparts. In summary, altered neutrophil responses are associated with both uncomplicated and complicated shigellosis.

L-rhamnose inhibits proliferation of murine splenocytes by the lipopolysaccharide and polysaccharide moiety of Shigella dysenteriae type 1 lipopolysaccharide.

Abstract: The induction of proliferation of murine splenocytes by lipopolysaccharide (LPS) of Shigella dysenteriae type 1 and its polysaccharide (PS) and lipid A fractions was investigated. The LPS-induced proliferation reached a maximum at a concentration of 30 ng/ml. The PS and lipid A induced proliferation of murine splenocytes at similar concentrations. Preincubation of murine splenocytes with varying concentrations of L-rhamnose blocked LPS- and PS-induced proliferation in a dose-dependent manner. The lipid A-induced stimulation, on the contrary, was not affected by preincubation of the cells with L-rhamnose. These data suggest that activation of splenocytes by LPS and PS is mechanistically different from that induced by lipid A and is presumably involved in the specific recognition of carbohydrate structures on LPS and PS.

Pathogenesis of bacterial colitis.

Abstract: developing fatal colorectal cancer. Aspirin seems to have immunostimulatory and tumoricidal properties in the colon' 2 thus accounting for the protection. It is these properties that may also make low dose aspirin useful in the chemoprevention of colorectal cancer in high risk patients.3 This is supported by the data of Manzano et al. Preneoplastic ulcerative colitis is associated with T lymphocyte immunosuppression and this may permit immunologically unchallenged malignant degeneration in a chronically inflamed mucosa. Aspirin may therefore be useful in the chemoprevention of colorectal cancer by augmenting the immune system thus destroying early tumours. This principle also applies in the oesophagus as there are several similarities between the carcinogenesis of colorectal and oesophageal cancer. Chronic inflammation may be a pre-neoplastic lesion in both organs4 while Barrett's oesophagus is also associated with immunosuppression.5 NSAIDs may thus be useful in the chemoprevention of both colorectal and oesophageal cancer6 in patients with pre-neoplastic disease. I agree with Choi and Zelig that longterm studies of low dose NSAID prophylaxis are warranted in patients receiving surveillance.