The effect of acupuncture in the treatment of young pigs with induced enteropathogenic Escherichia coli diarrhea was histopathologically evaluated by routine hematoxylin and eosin stain. Thirty two pigs weighed 4-5kg and aged 21days old were used in this study. The animals with diarrhea were treated with traditional acupuncture, or enrofloxacin. In the group treated with traditional acupuncture, acupoint GV1 (Jiaochao) was used and in the group treated with antibiotics, enrofloxacin was injected intramuscularly. Ten pigs were inoculated with E. coli, but were not treated and served as nontreated control group. At postinoculation day 6, all pigs of the acupuncture and antibiotic treated groups recovered from diarrhea. In the ascending and descending colons of the nontreated control group, severe infiltration of inflammatory cells in the lamina propria was observed and in the fundic stomach, destruction of the fundic gland architecture and necrotic lesions were observed, however, in the same sites of the acupuncture and antibiotics treated groups, the mucosae of the colon and stomach were relatively similar to those of the normal group. These results indicate that acupuncture treatment is effective in controlling induced E. coli diarrhea in pigs at its early stage.
Acupuncture ; Animals ; Colon/cytology/microbiology/pathology ; Diarrhea/therapy/*veterinary ; Escherichia coli Infections/therapy/*veterinary ; Gastric Mucosa/cytology/microbiology/pathology ; Intestinal Mucosa/cytology/microbiology/pathology ; Male ; Stomach/cytology/microbiology/pathology ; Swine ; Swine Diseases/*microbiology/therapy

OBJECTIVETo establish and evaluate a molecular diagnostic method for routine monitoring of four types of diarrheagenic Escherichia (E.) coli (DEC)and to study the distribution of four types of DEC isolated from diarrheal patients in Shanghai.

METHODSDEC-PCR standard operation procedure(SOP)had been developed for DEC detection and isolation, using the Statens Serum Institute (SSI) DEC PCR kits with multiplex PCR technique after verification tests on reference strains. Diarrhea specimens from 3 clinical hospitals in Shanghai were tested from June to September, 2012.

RESULTSSpecificity of the PCR kit was 100% by verification on the 26 DEC reference strains. A total number of 218 DEC isolates, including 181 fermented lactose and 37 unfermented lactose were identified from the 1887 stool specimens of diarrhea patients, with positive rate as 11.6%. The most common pathogen(54.1%, 118/218)was enteropathogenic E. coli(EPEC), followed by enterotoxigenic E. coli(ETEC, 41.3%, 90/218), enteroinvasive E. coli (EIEC, 4.1%, 9/ 218) and Shigatoxin-producing E. coli(STEC, 0.5%, 1/218)in addition to 18 Shigella isolates. ETEC dominated in diarrhea patients with foreign residency, as well as 1/3 were perinatal stage of neonatal ETEC of all diarrhea cases under the age of 5, while EPEC dominated in the Chinese diarrhea patients especially among young kids under the age of 2.

CONCLUSIONData was reliable after assessment on this molecular diagnostics and seperation procedures used for the routine monitoring on four types of DEC, while the diagnosis and reference ability of DEC regarding the laboratories net-working on food-borne pathogens need to be built up and improved.

Adolescent ; Adult ; China ; epidemiology ; Diarrhea ; diagnosis ; epidemiology ; microbiology ; Escherichia coli ; genetics ; isolation & purification ; Escherichia coli Infections ; diagnosis ; epidemiology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Pathology, Molecular ; Sentinel Surveillance

We evaluated the performance of a new chromogenic medium for detection of Clostridium difficile, chromID C. difficile agar (CDIF; bioMerieux, France), by comparison with BBL C. difficile Selective Agar (CDSA; Becton Dickinson and Company, USA). After heat pre-treatment (80degrees C, 5 min), 185 diarrheal stool samples were inoculated onto the two media types and incubated anaerobically for 24 hr and 48 hr for CDIF and for 48 hr and 72 hr for CDSA. All typical colonies on each medium were examined by Gram staining, and the gram-positive rods confirmed to contain the tpi gene by PCR were identified as C. difficile. C. difficile was recovered from 36 samples by using a combination of the two media. The sensitivity with CDIF 48 hr was highest (100%) and was significantly higher than that with CDIF 24 hr (58.3%; P<0.001), because samples with a low burden of C. difficile tended to require prolonged incubation up to 48 hr (P<0.001). The specificity of CDIF 24 hr and CDIF 48 hr (99.3% and 90.6%, respectively) was significantly higher than that of CDSA 48 hr and CDSA 72 hr (72.5% and 67.1%, respectively; P<0.001). CDIF was effective for detecting C. difficile in heat-pretreated stool specimens, thus reducing unnecessary testing for toxin production in non-C. difficile isolates and turnaround time.
Agar/chemistry ; Bacterial Proteins/genetics ; Bacteriological Techniques/*methods ; Chromogenic Compounds/chemistry/metabolism ; Clostridium difficile/genetics/*isolation & purification ; Culture Media/chemistry ; DNA, Bacterial/analysis/metabolism ; Diarrhea/microbiology/pathology ; Feces/*microbiology ; Humans ; Polymerase Chain Reaction ; Time Factors

In order to investigate the incidence, clinical and microbiologic characteristics of Clostridium difficile infection (CDI) in Korea, a prospective observational study was performed. From September 2008 through January 2010, all patients whose stool was tested for toxin assay A&B and/or C. difficile culture were studied for clinical characteristics. Toxin types of the isolates from stool were tested. The mean incidence of CDI per 100,000 patient-days was 71.6 by month (range, 52.5-114.0), and the ratio of CDI to antibiotic-associated diarrhea was 0.23. Among 200 CDI patients, 37.5% (75/200) was severe CDI based on severity score. Clinical outcome of 189 CDI was as followed; 25.9% (49/189) improved without treatment, 84.3% (118/140) achieved clinical cure and attributed mortality was 0.7% (1/140) with the treatment. Recurrence rate was 21.4% (30/140) and cure without recurrence was 66.4% (93/140). The most common type of toxin was toxin A-positive/toxin B-positive strain (77.5%), toxin A-negative/toxin B-positive strains or binary toxin-producing strains comprised 15.4% or 7.1%, respectively. In conclusion, the incidence of CDI in Korea is a little higher than other reports during the non-epidemic setting. We expect that the change of epidemiology and clinical severity in CDI can be evaluated based on these results.
Aged ; Bacterial Proteins/analysis ; Bacterial Toxins/analysis ; Clostridium Infections/*epidemiology/physiopathology ; Clostridium difficile/*isolation & purification/*pathogenicity ; Diarrhea/epidemiology/microbiology ; Enterocolitis, Pseudomembranous/*epidemiology/microbiology/pathology ; Enterotoxins/analysis ; Feces/microbiology ; Female ; Hospitals ; Humans ; Incidence ; Male ; Metronidazole/therapeutic use ; Middle Aged ; Prospective Studies ; Recurrence ; Republic of Korea/epidemiology ; Treatment Outcome ; Vancomycin/therapeutic use

OBJECTIVETo explore the effects of Huoxiang Zhengqi liquid (HXZQ) on enteric mucosal immune responses in mice with Bacillus dysenteriae and Salmonella typhimurium induced diarrhea (BSD).

METHODMice were randomly divided into four groups with 10 mice in each group: control group (control), BSD group, Huoxiang Zhengqi liquid treated BSD groups at high dosage and low dosage (HXZQ high, HXZQ low). HXZQ was administrated from the day of diarrhea induction at dosage of 5.21 g kg(-1) and 0.52 g kg (-1) respectively. Peyer's patch and periphery lymphocytes were prepared for flow cytometry, and level of TNF-alpha in periphery and enteric tissue homogenate were determined with ELISA. Student's t-test was used for statistics.

RESULTMice in BSD group started showing continuous diarrhea at the day of induction till the fourth day when the mice were sacrificed. Diarrhea in the mice of HXZQ high and low groups lasted for 36 and 54 h respectively. There were more CD4+ and CD8+ cells in periphery, less CD4+ cells in peyer's patch in BSD mice comparing to normal mice. In peyer's patch, there were more CD8+ cells in mice in HXZQ high and low groups and more CD4+ in mice in HXZQ high group. Higher level TNF-alpha in periphery and intestinal tissue homogenate in BSD group were observed. Mice in HXZQ high group showed the decreased level TNF-alpha in periphery and enteric tissue homogenate.

CONCLUSIONThe immune regulation on peyer's patch CD4+ and CD8+ cells and suppression on TNF-alpha level in enteric homogenate might partially explain the effect of HXZQ on improvement of BSD.

Animals ; CD4-CD8 Ratio ; Colon ; immunology ; metabolism ; pathology ; Diarrhea ; immunology ; metabolism ; microbiology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Dysentery, Bacillary ; immunology ; metabolism ; microbiology ; Immunity, Mucosal ; drug effects ; Intestinal Mucosa ; immunology ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Peyer's Patches ; drug effects ; immunology ; pathology ; Plants, Medicinal ; chemistry ; Random Allocation ; Salmonella Infections ; immunology ; metabolism ; microbiology ; Salmonella typhimurium ; immunology ; Shigella dysenteriae ; immunology ; T-Lymphocyte Subsets ; drug effects ; immunology ; pathology ; Tumor Necrosis Factor-alpha ; blood ; metabolism