Surveillance of Antimicrobial Susceptibility Patterns among Shigella Species Isolated in China during the 7-Year Period of 2005-2011

BACKGROUND: Shigella is a frequent cause of bacterial dysentery in the developing world. Treatment with antibiotics is recommended for shigellosis, but the options are limited due to globally emerging resistance. This study was conducted to determine the frequency and pattern of antimicrobial susceptibility of Shigella in China. METHODS: We studied the antimicrobial resistance profiles of 308 Shigella spp. strains (260 S. flexneri, 40 S. sonnei, 5 S. boydii, and 3 S. dysenteriae) isolated from fecal samples of patients (age, from 3 months to 92 yr) presenting with diarrhea in different districts of Anhui, China. The antimicrobial resistance of strains was determined by the agar dilution method according to the CSLI guidelines. RESULTS: The most common serogroup in the Shigella isolates was S. flexneri (n=260, 84.4%), followed by S. sonnei (n=40, 13.0%). The highest resistance rate was found for nalidixic acid (96.4%), followed by ampicillin (93.2%), tetracycline (90.9%), and trimethoprim/sulfamethoxazole (80.8%). Among the isolates tested, 280 (91.0%) were multidrug resistant (resistant to > or =2 agents). The most common resistance pattern was the combination of ampicillin, tetracycline, and trimethoprim/sulfamethoxazole (70.8%). Resistance to ampicillin and tetracycline were more common among S. flexneri than among S. sonnei isolates. CONCLUSIONS: S. flexneri is predominant in Anhui, China, and its higher antimicrobial resistance rate compared with that of S. sonnei is a cause for concern. Continuous monitoring of resistance patterns is necessary to control the spread of resistance in Shigella. The recommendations for antimicrobial treatment must be updated regularly based on surveillance results.

Diarreia associada a Shigella em crianças e sensibilidade a antimicrobianos / Diarrhea associated with Shigella in children and susceptibility to antimicrobials

OBJETIVO: Avaliar a distribuição e suscetibilidade a antimicrobianos de Shigella isolada de crianças com diarreia aguda e sem diarreia em Teresina (PI). MÉTODOS: Quatrocentas crianças com idade até 60 meses foram estudadas. Fezes foram coletadas de todos os pacientes entre janeiro de 2004 e agosto de 2007. Shigella foi identificada por métodos convencionais e antibiograma e pesquisa de β-lactamase de espectro ampliado (ESBL) foram realizados por difusão em ágar. RESULTADOS: Shigelose foi detectada apenas em crianças com diarreia aguda (26/250; 10,4%), especialmente naquelas entre 6 e 24 meses de idade e nos meses chuvosos. Shigella foi suscetível a ceftriaxona, ciprofloxacina e ácido nalidíxico. Mais da metade das amostras foram resistentes a sulfametoxazol-trimetoprim e ampicilina. ESBL não foi detectada. CONCLUSÕES: S. flexneri é comum em Teresina. A resistência a ampicilina e sulfametoxazol-trimetoprim é preocupante, pois estas drogas são amplamente utilizadas na prática e sulfametoxazol-trimetoprim ainda é recomendada para tratamento de crianças com suspeita de shigelose.

OBJECTIVE: To evaluate the distribution and susceptibility to antimicrobials of Shigella isolated from children with acute diarrhea and without diarrhea in Teresina, state of Piauí, Brazil. METHODS: Four hundred children aged up to 60 months were studied. Stools were collected from all the patients between January 2004 and August 2007. Shigella was identified by conventional methods and antibiogram and extended-spectrum β-lactamase (ESBL) were performed by agar diffusion. RESULTS: Shigellosis was only detected in children with acute diarrhea (26/250; 10.4%), especially in those aged from 6 to 24 months and in the rainy months. Shigella was susceptible to ceftriaxone, ciprofloxacin and nalidixic acid. More than half of the strains were resistant to sulphametoxazole-trimethoprim and ampicillin. ESBL was not detected. CONCLUSIONS: S. flexneri is common in Teresina. The resistance to ampicillin and sulphametoxazole-trimethoprim gives cause for concern, as these drugs are widely used in practice and sulphametoxazole-trimethoprim is also recommended for treating children suspected of having shigellosis.

Quinolone-resistant Shigella flexneri Isolated in a Patient Who Travelled to India

We report a recent case in which ciprofloxacin-resistant Shigella flexneri was isolated from a 23-yr-old female patient with a history of travel to India. Prior to her admission to our internal medicine department, she experienced symptoms of high fever and generalized weakness from continuous watery diarrhea that developed midway during the trip. S. flexneri was isolated from the stool culture. Despite initial treatment with ciprofloxacin, the stool cultures continued to show S. flexneri growth. In the susceptibility test for antibiotics of the quinolone family, the isolate showed resistance to ciprofloxacin (minimum inhibitory concentration [MIC], 8 microg/mL), norfloxacin (MIC, 32 microg/mL), ofloxacin (MIC, 8 microg/mL), nalidixic acid (MIC, 256 microg/mL), and intermediate resistance to levofloxacin (MIC, 4 microg/mL). In molecular studies for quinolone resistance related genes, plasmid borne-quinolone resistance genes such as qnrA, qnrB, qnrS, aac(6')-Ib-cr, qepA, and oqxAB were not detected. Two mutations were observed in gyrA (248C-->T, 259G-->A) and 1 mutation in parC (239G-->T). The molecular characteristics of the isolated S. flexneri showed that the isolate was more similar to the strains isolated from the dysentery outbreak in India than those isolated from Korea.

Ceftriaxone resistant Shigella Flexneri, an emerging problem.

Shigellosis is a disease of public health importance in developing countries. It may cause self-limited diarrhea to severe dysentery. Emergence of multi drug resistant (MDR) strains is a growing concern globally. Ceftriaxone and ciprofloxacin are the drugs of choice for MDR cases. Here, we report a case of MDR Shigella flexneri from an immunocompromised patient. The strain was resistant to ceftriaxone [minimum inhibitory concentration (MIC) ≥ 64 μg/ml], limiting the treatment option. Simultaneously, the strain was also found to be resistant to ciprofloxacin (MIC ≥ 4 μg/ml). However, it was susceptible to ceftazidime (MIC 4 μg/ml). This is the first case of ceftriaxone resistant Shigella spp. reported from our hospital.

Surveillance of antibiotic susceptibility pattern among shigella flexneri strain isolated in Nagpur district during three years period, January 2009-January 2012.

Objective : In the few cases of childhood dirrhea that require the antimicrobial therapy, the correct choice of the drug depends on detailed previous knowledge of local strains and pattern of antimicrobial resistance. Shigellosis is one of the most improtant examples of this group of intestinal infections. In order to establish such parameters in Nagpur city, this study was carried out to determine the antimcrobial resistance profile of Shigella flexneri isolated from patients suffering from diahhrea admitted to Various hoapitals in Nagpur district, India. Materials and Methods: The study included 110 stool samples collected from patients during the 3 year period. All the isolates were characterized and confirmed by VITEK® 2 GN ID cards and antimicrobial susceptibility was tested by VITEK® 2 AST test cards. Results: We received 73 positive cultures of S. flexneri out of 110 stool samples during three year periods of January 2009 to January 2012. S. flexneri strains presented a high resistance rate to Ampicillin (100%), Chloramphenicol (76.71%), Trimethoprime-sulfamethaxazole (TMP-SMZ) (68.49%) and low resistance to third- and fourth-generation Cephalosporin. None of the isolates was found to be resistant to Ciprofloxacin (MIC ≥ 4), Norfloxacin (MIC ≥12), and Nalidixic acid (MIC ≥30). Conclusion: Our results provide data on antimicrobial resistance to choose a proper antibiotic for the treatment of Shigellosis in our country. According to current findings, Quinolones and Cephalosporins are the drug of choice for the diarrheic patients. In conclusion, systematic monitoring is needed to identify changes in the antimicrobial resistance.

Epidemiological characterization of resistance and PCR typing of Shigella flexneri and Shigella sonnei strains isolated from bacillary dysentery cases in Southeast Brazil

Shigella spp are Gram-negative, anaerobic facultative, non-motile, and non-sporulated bacilli of the Enterobacteriaceae family responsible for "Shigellosis" or bacillary dysentery, an important cause of worldwide morbidity and mortality. However, despite this, there are very few epidemiological studies about this bacterium in Brazil. We studied the antibiotic resistance profiles and the clonal structure of 60 Shigella strains (30 S. flexneri and 30 S. sonnei) isolated from shigellosis cases in different cities within the metropolitan area of Campinas, State of São Paulo, Brazil. We used the following well-characterized molecular techniques: enterobacterial repetitive intergenic consensus, repetitive extragenic palindromic, and double-repetitive element-polymerase chain reaction to characterize the bacteria. Also, the antibiotic resistance of the strains was determined by the diffusion disk method. Many strains of S. flexneri and S. sonnei were found to be multi-resistant. S. flexneri strains were resistant to ampicillin in 83.3 percent of cases, chloramphenicol in 70.0 percent, streptomycin in 86.7 percent, sulfamethoxazole in 80.0 percent, and tetracycline in 80.0 percent, while a smaller number of strains were resistant to cephalothin (3.3 percent) and sulfazotrim (10.0 percent). S. sonnei strains were mainly resistant to sulfamethoxazole (100.0 percent) and tetracycline (96.7 percent) and, to a lesser extent, to ampicillin (6.7 percent) and streptomycin (26.7 percent). Polymerase chain reaction-based typing supported the existence of specific clones responsible for the shigellosis cases in the different cities and there was evidence of transmission between cities. This clonal structure would probably be the result of selection for virulence and resistance phenotypes. These data indicate that the human sanitary conditions of the cities investigated should be improved.

Study of isolation and determination of antibiotic resistance, plasmid profiles, protein bands and phenotypic virulence of Shigella flexneri strains

Shigellosis is an acute gastroenteritis caused by Shigella species, including S. dysenteriae, S. flexneri, S. boydi and S. sonnei.The purpose of this study was to isolate and to determine antibiotic resistance, plasmid profile and protein b and s by SDS-PAGE, and also phenotypic virulence by Congo red dye among S. flexneri strains. The isolated bacteria were identified by use of st and ard bacterial and biochemical methods. Antibiotic susceptibility tests were performed according to kirby-Bauer method. Plasmids were isolated by alkaline lysis method. Serological reactions were detected by slide agglutination tests with both polyclonal and monoclonal antisera kits. Virulent strains were isolated on a TSA plate containing Congo red dye. From 350 isolated Shigella spp. 142 [40.57%] were S. flexneri. Of 350 patients 41% were female and 59% male. In 100 S. flexneri isolates, resistance rates to tetracycline, ampicillin, trimethoprim-sulfometoxazol and cephalexin were 95%, 91.3%, 85.6% and 70.3% respectively. All of isolates were sensitive to ciprofloxacin. Most isolates contained multiple plasmid b and s [1-5 plasmid's b and s]. A total of 11 distinct plasmid profile patterns were identified. The results of our study showed that serotype 2 was the most common isolated serotype of S. flexeneri [39%]. In this study, 46% of S. flexneri were Congo red positive and haemolysin positive on blood agar plates. A 120 KDa protein b and was detected on electrophoresis gel. These data showed, pathogenicities of all S. flexneri isolates are not similar. Health care can prevent bacterial diarrhea due to shigella. Therefore laboratories are recommended to be more concerned about isolation of these bacteria. Congo red dye binding test is cheap, simple and rapid, so, it can be used to determine the virulence properties of S. flexneri. Antibiogram tests are recommended to prevent antibiotic resistance

Experimental study on the antibacterial effect of origanum volatile oil on dysentery bacilli in vivo and in vitro / 华中科技大学学报（医学）（英德文版）

To observe the germistatic and germicidal effects of origanum volatile oil (OVI) on the dysentery bacteria, the abdominal cavity of mice was infected with Shigella sonne (Sh. sonnei) and Shigella flexneri (Sh. flexneri) F2a. After OVI was given to the mice via gastric lavage, the effects of OVI on the infected mice were observed. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) for dysentery bacteria were determined in vitro. The results showed that origanum volatile oil showed obvious protective effect on mice infected with Sh. sonnei and Sh. flexneri F2a and it had germistatic and germicidal effects on dysentry bacteria. We are led to conclude that origanum volatile oil is an effective medicine against the infection of dysentery bacteria.

Antibacterial effects of Carica papaya fruit on common wound organisms

The purpose of the study was to investigate antibacterial activity of ripe and unripe Carica papaya on selected micro-organisms. Cultures of micro-organisms were routinely maintained in nutrient agar slants at 4 degrees C. Extracts of immature, mature and ripe Carica papaya fruit were obtained by separately grinding factions of the epicarp, endocarp and seeds and filtering them through gauze. Sensitivity tests were conducted by adding 0.06 ml of extract to agar wells (6 mm diameter) prepared from 20 ml agar seeded with 10(6) cells/ml suspension of one of the eight organisms per plate. The inoculated plates were allowed to equilibrate at 4 degrees C for 1 hour, incubated at 37 degrees C for 24 hours, and zones of inhibition measured in millimetres. Anti-bacterial activity was expressed in terms of the radius of zone of inhibition. Seed extracts from the fruit showed inhibition in the following order: B cereus > E coli > S faecalis > S aureus > P vulgaris > S flexneri. No significant difference was found in bacterial sensitivity between immature, mature and ripe fruits. No inhibition zone was produced by epicarp and endocarp extracts. Carica papaya seeds contain anti-bacterial activity that inhibits growth of gram-positive and gram-negative organisms. Observed activity was independent of stage of fruit maturity. Carica papaya has antibacterial effects that could be useful in treating chronic skin ulcers to promote healing

Integrones y cassettes genéticos de resistencia a antimicrobianos en cepas de Shigella flexneri / Integrons and antimicrobial resistance gene cassettes in Shigella flexneri strains

BACKGROUND: The resistance of Shigella flexneri to antimicrobial agents can be associated to the presence of integrons that may contain and express antimicrobial resistance gene cassettes. AIM: To study antimicrobial resistance and the presence of integrons and antimicrobial gene cassettes in Shigella flexneri strains. MATERIAL AND METHODS: In vitro susceptibility to 27 antimicrobials was studied in twenty four Shigella flexneri strains isolated from stools. The presence of integrons class 1, 2 and 3 and antimicrobial resistance gene cassettes was investigated by polymerase chain reaction (PCR) using specific primers for each gene. RESULTS: Most strains were resistant to one of the following antimicrobials: ampicillin, sulphonamide, trimethoprim, tetracycline, streptomycin, sulfamethoxazole-trimethoprim or chloramphenicol. Twenty nine percent were simultaneously resistant to all these antimicrobials. Integrons class 1 and 2 were found in 19 strains (79 per cent). Class 3 integrons were not found. Gene cassettes dfrA1 and ant(3")I were associated to integrons class 2 in most strains (15/20, 75 per cent). Genes cat, tetB and blarTEM were detected in 18/24 (75 per cent), 7/24 (29 per cent) and 4/24 (17 per cent) of the strains, respectively and were not associated to any of the studied integrons. Genes that codify enzymes AAC(6')Ib and APH(3')VI were not detected in any strain. CONCLUSIONS: The high frequency of integrons found in the studied strains, could partly explain the increasing antimicrobial resistance of Shigella flexneri strains, isolated in Chile.

Shigella isolation in Vellore, south India (1997-2001).

Shigellosis has been a major cause of dysentery for many years at Vellore, south India. In the last two years the number of Shigella being isolated from samples of faeces from patients with diarrhoea has decreased (5% isolation rate in 1997 to 3.9% in 2001), although the microbiological methods and media used have not changed. Also, the nalidixic acid (NA) resistance has increased for S. sonnei (now 94%). This is noteworthy, since NA has been recommended for the empirical treatment of patients suspected to have shigellosis and this concept needs to be reconsidered based on available data.

Antimicrobial resistance of Shigella spp isolated from diarrheal patients between 1989 and 1998 in Vietnam.

Shigellosis is an important cause of infectious diarrhea in Vietnam, caused mainly by Shigella flexneri. This study provides for the first time in the international literature, data on the development of antimicrobial resistance in Shigella between 1988 to 1998, including data reported to the National Program for Surveillance of Antimicrobial Resistance (NPSAR). Our studies show that about 80% of the Shigella strains tested were resistant to ampicillin, chloramphenicol, oxytetracycline, trimethoprim and sulfonamides. This combination of drugs was also the most common antibiogram among multiple-resistant S. flexneri (57%). Resistance to tetracyclines, sulfonamides and, in particular trimethoprim (p<0.001), increased during the study period. Our findings indicate that tetracyclines and co-trimoxazole (a combination of a sulfonamide and trimethoprim), which are recommended and commonly used drugs for the treatment of shigellosis in Vietnam, may have limited therapeutic effect. In contrast to neighboring countries, low percentages of resistance were found to nalidixic acid and norfloxacin (3-5%) and no resistance was found to ciprofloxacin, indicating that nalidixic acid with its low cost and safety in children could be recommended for the treatment of shigellosis. The NPSAR provides a useful picture of the levels and development of antimicrobial resistance in Vietnam and should receive continued support for further improvement by increasing the number of provinces covered, the numbers of isolates tested from rural areas, and the communication of results to medical practitioners and others prescribing and/or selling antimicrobials.

Multirresistência a antimicrobianos mediada por plasmídios R em cepas de Shigella flexneri isoladas no nordeste do Brasil / Multiple antibiotic resistance mediated by R plasmid in Shigella flexneri strains isolated in the northeast of Brazil

Em amostras de S. flexneri, isoladas no período de 1989 a 1993, foi estudado o mecanismo molecular que mediava a multirresistência. Foram utilizadas no estudo 26 amostras de S. flexneri. Estas amostras foram submetidas a teste de sensibilidade a antimicrobianos, experimentos de conjugação e extração de plasmídios. Com relação ao padrão de resistência a antimicrobianos, observou-se que todas as amostras de S. flexneri eram resistentes a pelo menos três antimicrobianos. Das 26 amostras de S. flexneri doadoras submetidas ao processo de conjugação, 34,6% (9 amostras) resultaram em uma freqüência variável de transconjugantes. Das amostras que conjugaram, 100% transferiram o fator de resistência relacionado à ampicilina; sendo que em todas as transconjugantes foi evidenciado apenas um plasmídio de 23,1Kb. Este plasmídio, encontrado em todas as amostras de Shigellas, caracterizou-se como o portador de marca de resistência para ampicilina.

In Shigella strains were studied the molecular mechanism that mediated the multiply antibiotic-resistance. Twenty-six strains of Shigella flexneri were utilised in this investigation. These strains were submitted to disk diffusion test, mating experiments and plasmid isolation. In relation to antibiotics resistance standard it was observed that all Shigella flexneri strains were resistant to at least, three antibiotics tested. From twenty-six Shigella flexneri strains donors submitted to conjugation process, 34.6% (nine strains) resulted in variable frequency of transconjugants. From strains that conjugated, 100%, transferred the resistance factor acquainted with ampicillin. Being that, in all transconjugants which were observed, just one plasmid with 23.1 Kb was evidenced. This plasmid found in all strains was characterised as the cause of resistance to ampicillin.

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

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.

Changing antibiotic sensitivity patterns in shigellosis.

Over five and a half months stool cultures were performed in 154 patients admitted with suspected bacillary dysentery to the Professorial Paediatric Unit of the Teaching Hospital, Galle. Thirty one patients had a positive stool culture; Shigella dysenteriae type 1 was isolated in 23 patients, and S flexneri in 8. In both groups the antibiotic sensitivity pattern was uniform, the only difference being S dysenteriae type 1 being resistant to nalidixic acid while S flexneri was sensitive. This antibiotic sensitivity pattern was different to that reported from Colombo during the same period. Complications were observed in some patients with S flexneri infection.

Efficacy of aqueous extract of garlic & allicin in experimental shigellosis in rabbits.

The aqueous extract of garlic (Allium sativum) and allicin both showed significant in vitro antibacterial activity against isolates of multiple drug-resistant Shigella dysenteriae 1, Sh. flexneri Y, Sh. sonnei and enterotoxigenic Escherichia coli. The minimum inhibitory concentrations of the aqueous extract and allicin against Sh. flexneri Y were 5 and 0.4 microliters/ml, respectively. The two agents also showed promising in vivo antibacterial activity against Sh. flexneri Y when tested in the rabbit model of experimental shigellosis, fully curing the infected rabbits within 3 days. On the contrary, 4 of the 5 rabbits in the control group died within 48 h. The rectal swab of rabbits of the experimental groups became free of the challenge bacteria on the second day of treatment. The antibacterial activity against the challenge strain was observed in the sera of the treated rabbits with 30-60 min of administration of the agents. The LD50 values of the aqueous extract and allicin in mice were 173.78 ml/kg and 204.17 microliters/kg of body weight, respectively. At the therapeutic dose, the two agents did not show any adverse effects on the standard biochemical profile of blood.