Inhibitory Effect of Sodium Citrate, Sodium Nitrite and Cinnamaldehyde on Biofilm-forming Escherichia coli O157:H7

The development of biofilms by the foodborne pathogens attached to surfaces in the food processing environments results in the deterioration of products, persistence of pathogenic bacteria and transmission of food-associated diseases. In addition, biofilms are more resistant to antimicrobials than their planktonic counterparts which make their elimination from food and the food processing facilities a great challenge. This study aim was to determine the inhibitory effect of food additives on biofilm forming Escherichia coli O157:H7. The isolate obtained was subjected to Gram’s staining and various biochemical identifications and later confirmed by latex agglutination test. Biofilm formation potential was done on Congo red media and the confirmed biofilm former was subjected to biofilm formation at 10℃ and 37℃ for 168hrs. Antimicrobial susceptibility testing, MIC, MBC, and antibiofilm effect was determined following CLSI 2017 guideline. The highest zone of growth inhibition of 31 mm was exhibited by cinnamaldehyde, sodium nitrite with 26 mm and sodium citrate with 13 mm. The MIC 2.5 mg/mL was recorded for sodium citrate, 0.25 mg/mL for sodium nitrite and 0.125 μl/mL for cinnamaldehyde. Strong biofilm was formed at 37 ℃ with 7.82 x 109 CFU/mL viable cells at 168hrs while 6.79 x 109 CFU/mL were obtained at 10 ℃. All the three additives showed antibiofilm effect (at 10℃ and 37℃), cinnamaldehyde exhibited 70%-90.1%, sodium nitrite; 70%-88.2% inhibition and sodium nitrite; 75%-88% inhibition respectively. This study showed that sodium citrate, sodium nitrite and cinnamaldehyde exerted strong antimicrobial and antibiofilm properties indicating their potential as good preservatives.

Prevalence of non-O157 Shiga Toxin-producing E. Coli in Children and Calves in Al- Muthanna Province, Iraq

The present study focus on non-O157 Shiga toxin-producing E. Coli (STEC), included a bacteriological study was subjected to provide additional information for non-O157 STEC prevalence in children and calves. Isolation by using selective culturing media (CHROMagar STEC and CHROMagar O157) from 127 children suffering from diarrhea and 133 calves in Al- Muthanna province. Characterization depends on culturing positive colony on MacConkey agar and Levin’s Eosin Methylene blue agar, staining single colony from the growth by gram stain, biochemical tests; Indole, the Methyl Red, Voges-Proskauer, Citrate test, Oxidase, Catalase, Urease, Motility, Kligler Iron and Api-20E, were done to confirm a diagnosis of non-O157 STEC, The reliable isolation as non-O157 STEC serotyping by specific latex agglutination test for the target non-O157 STEC (big six) serogroup (O26, O45, O103, O111, O121 and O145). The current study showed the prevalence of non-O157 STEC was 20 of out 127 (15.73%) in samples collected from children and 27 / 133 (20.30%) in calves samples in conclusion the Non-O157 STEC is an important cause of diarrhea in children, and calves; finally, the calves play an important reservoir for Non-O157 STEC.

Predictive Value of Combination of D-dimer and Fibrinogen in Diagnosing Acute CVST

Background: Cerebral venous sinus thrombosis (CVST) is an uncommon subtype of stroke, and the role of D-dimer and fibrinogen in early diagnosis of CVST has been studied with varying results. The present study aims to study the role of the combination of D-dimer and fibrinogen in early diagnosis of acute CVST. Methods: Forty consecutive confirmed acute CVST cases admitted at a tertiary care center were recruited for the study. D-dimer and fibrinogen were assessed by a rapid semiquantitative latex agglutination assay. Results: Out of the 40 CVST patients, 21 (52.50%) were females. The mean age of the patients was 37.58 years ± 19.17 years. Common clinical features were headache ( N = 38 [95%]), papilloedema ( N = 15 [37.5%]), and seizures ( N = 12 [30%]). Although the sensitivity (75%) of the combination of D-dimer and fibrinogen assay was lower than that of D-dimer or fibrinogen alone, the specificity and positive predictive value (75% each) was higher. Conclusions: The combination of D-dimer and fibrinogen testing may aid in an early diagnosis of acute CVST and in better management.

Exposure to Toxoplasma gondii in Asian Elephants (Elephas maximus indicus) in Thailand

Toxoplasma gondii is the causative agent of toxoplasmosis in humans and various animal species worldwide. In Thailand, seroprevalence studies on T. gondii have focused on domestic animals, and information on infections in Asian elephants (Elephas maximus indicus) is scarce. This study was conducted to determine the seroprevalence of T. gondii infection in archival sera collected from 268 elephants living in Thailand. The serum samples were analyzed for anti-T. gondii immunoglobulin G antibodies using the latex agglutination test (LAT) and indirect enzyme-linked immunosorbent assay (iELISA) based on T. gondii lysate antigen (TLA-iELISA) and recombinant T. gondii dense granular antigen 8 protein (TgGRA8-iELISA). The prevalence of antibodies against T. gondii was 45.1% (121/268), 40.7% (109/268), and 44.4% (119/268) using LAT, TLA-iELISA, and TgGRA8-iELISA, respectively. Young elephants had a higher seropositivity rate than elephants aged >40 years (odds ratio = 6.6; p < 0.001; 95% confidence interval: 2.9–15.4). When LAT was used as the reference, TLA-iELISA and TgGRA8-iELISA showed a substantial (κ = 0.69) and moderate (κ = 0.42) agreement, respectively. Although our findings suggest the widespread exposure of Asian elephants to T. gondii in Thailand, the source of infection was not investigated. Therefore, investigation of the predisposing factors associated with toxoplasmosis is necessary to identify the potential risk factors for infection.

Prevalence and the risk factors for the carriage of beta-haemolytic streptococci among women visiting a tertiary care hospital in South India

Streptococci are gram positive cocci arranged in chains and are part of normal flora of humans and animals. The present study is carried out to determine the prevalence and risk factors for the carriage of beta-haemolytic streptococci (BHS) among women visiting Dr. VRK Women’s Teaching Hospital & Research Centre, Hyderabad. Vaginal swabs were collected from 250 patients attending outpatient department (OPD) of Dr. VRK Women’s Teaching hospital. Swabs were inoculated onto 5% sheep blood agar plates and incubated for 24 h at 37°C in a candle jar. BHS isolates were phenotypically identified by standard microbiological techniques, all the isolates presumptively identified as BHS were tested for Bacitracin susceptibility. Sensitive isolates were presumptively identified as GAS and resistant isolates were identified as non-group A BHS (NGABHS). Presumptively identified GAS & NGABHS isolates were serogrouped by Lancefield grouping using a commercially available latex agglutination test. BHS were isolated from 12.4% of samples. As many as 12 BHS isolates were identified as GAS and 19 were identified as NGABHS. Ten of nineteen were identified as group B (GBS), 4 (12.9%) were identified as group C (GCS) and 5 (16.12%) were identified as group G (GGS). Among six clinical groups, the prevalence of GAS is highest i.e. 7.5% in female patients visiting Gynaecology OPD with history of white discharge. Prevalence of NGABHS was more among post insertion (18%) IUCD group compared to pre insertion (8%) IUCD group. GBS were isolated from 7% of samples from IUCD group and 4% of samples from prostitutes.This study reports the prevalence of BHS among women visiting a tertiary care hospital in Hyderabad. This study also identified certain risk factors such as IUCD usage and working as a FSW are associated with the increased prevalence of NGABHS especially GBS.

Prevalence and antibiotic susceptibility of Streptococcus pyogenes isolated from pyoderma in a tertiary care hospital, Hyderabad, South India

Pyoderma is a common acute superficial bacterial skin infection which is highly contagious. In the great majority of cases, pyoderma is caused by , , or both. The present study was carried out to determine the prevalence and antibiotic susceptibility of isolated from pyoderma in Dr. VRK Women’s Teaching hospital. Swabs or pus samples were collected from 250 patients attending Dermatology, outpatient department (OPD) of Dr. VRK Women’s Teaching hospital. Samples were inoculated onto 5% sheep blood agar plates and incubated for 24 h at 37°C in a candle jar. BHS isolates were phenotypically identified by standard microbiological techniques, all the isolates presumptively identified as BHS were tested for Bacitracin susceptibility. Presumptive identification of a strain as a Group A Streptococcus (GAS) was also made by PYRase test. Presumptively identified GAS isolates were serogrouped by Lancefield grouping using a commercially available latex agglutination test. isolates were subjected to antimicrobial susceptibility testing by Kirby-Bauer disc diffusion method. BHS were isolated from 30% of samples. Prevalence of BHS was more among 0-10 years age group (38%). BHS were isolated more frequently from males (38.8%). were isolated from 52 (20.8%) samples. All 52 isolates were found to be susceptible to Penicillin G, amoxicillin, ceftriaxone, azithromycin and vancomycin. Erythromycin and clindamycin showed good activity with sensitivity rates of 92.3% & 96.1%, respectively. Resistance to tetracycline (59.6%) and chloramphenicol (23.1%) was commonly seen in . This study reports the prevalence and antibiotic susceptibility of isolated from pyoderma in Dr. VRK Women’s Teaching hospital. Results of this study suggests the peak incidence of pyoderma in children aged 0 to 10 years and male preponderance. Our study also reports high prevalence of tetracycline and chloramphenicol resistance in .

Evaluation of Real-Time Polymerase Chain Reaction in Culture-Negative Cerebrospinal Fluid Samples of Bacterial meningitis Patients

Background: Meningitis is a rigorous childhood disease with high morbidity and mortality. It is the main cause of under five mortality in India. Mainly three bacteria Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae are responsible. In low economic set up country like India, documented bacterial meningitis mainly depend on gram staining, cerebrospinal fluid (CSF) culture results or latex agglutination test resulting in less number of positive due to the prior antimicrobial intake which affects culture and latex agglutination test results. This study was taken up rapid and accurate molecular method like RT PCR to diagnose bacterial meningitis in culture-negative CSF samples. Materials and Methods: Fifty culture-negative CSF samples from suspected cases of bacterial meningitis were examined by real-time polymerase chain reaction (real-time PCR) for the presence of lytA, bexA, and ctrA genes specific for Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis respectively. Results: Positive real-time PCR results for Streptococcus pneumoniae were detected in 36 (72%) of culture-negative CSF samples while 10% positive results for Haemophilus influenzae type b. Nine (18%) samples were negative by real-time PCR for all tested organisms. Conclusion: The use of molecular techniques as real-time PCR can provide a valuable addition to the proportion of diagnosed cases of bacterial meningitis especially in settings with high rates of culture-negative results.

Antibiotic Resistances and Molecular Characteristics of Clostridioides difficile in ICUs in a Teaching Hospital From Central South China

Clostridioides (C.) difficile is a major healthcare-associated pathogen inducing infectious diarrhea. Approximately 25–33% of patients with antibiotic-associated diarrhea (AAD) and 90% of patients with pseudomembranous enteritis are caused by C. difficile infection (CDI). Stool samples were collected from hospitalized adults with presumptive AAD in four nonneonatal intensive care units (ICUs). Diagnosis of CDI was based on both clinical symptoms and laboratory results. The stool specimens were transferred onto CDIF (C. difficile agar), and C. difficile was finally confirmed by the latex agglutination test. Toxin-producing genes tcdA (A), tcdB (B), and cdt (CDT) were detected by PCR, and all isolates were performed multilocus sequence typing analysis. The antibiotic susceptibility of C. difficile isolates was assessed by the agar dilution method. A total of 184 C. difficile were isolated from 857 specimens in our study, the isolation rate of C. difficile was 21.5% (184/857). The 184 C. difficile were isolated from 179 patients, among these 115 patients were toxin-positive, giving the incidence of CDI being 58.0/10,000 patient days in the four ICUs. Among these 115 toxin-positive C. difficile isolates, 100 (87.0%) isolates produced two toxins (A+B+CDT-), three (2.6%) isolates were A+B+ with binary toxin-producing (A+B+CDT+), and 12 (10.4%) isolates only produced one toxin (A-B+CDT-). A total of 27 sequencing types (STs) were obtained. The most prevalent was ST3 (34 isolates), followed by ST39 (27 isolates), ST54 (19 isolates), ST26 (16 isolates), ST35 (15 isolates), and ST2 (13 isolates). All the ST26 isolates were nontoxigenic. Meanwhile, five STs were newly discovered. Although multidrug resistance was present in ≥50% of these C. difficile isolates, all of them were susceptible to tigecycline, fidaxomicin, metronidazole, and vancomycin. In conclusion, C. difficile isolates producing two toxins (A+B+CDT-) were dominant in our hospital. The most prevalent was ST3, and all ST26 isolates were NTCD. Although multidrug resistance was present in ≥50% of the C. difficile isolates, metronidazole, tigecycline, fidaxomicin, and vancomycin were still effective treatments for CDI in our hospital.

DETECTION OF TOXOPLASMOSIS IN CATS AND SHEEP

Thirty-three fecal samples from cats were examined for the presence of Toxoplasma oocysts, and another 33 serum samples from these cats were subjected for Latex agglutination test & indirect immunofluorecent antibody test. Also 80 serum samples from ewes were subjected to the same serological tests. The study indicated that the prevalence of Toxoplasma oocysts in cats was 27.3%. Higher rates of antibody titer (68%) were observed in cats tested with latex test. Infection in young cats was higher than in adults. Sixty percentage of ewes were sero-positive with Latex test, but only 35% were sero-positive with IFAT, higher prevalence of antibody titers was observed in sheep from the three locations of Iraq. Ewes that had recurrent abortion showed higher prevalence in both tests than non aborted ewes.

Long-term Storage of Bacterial Isolates by Using Tryptic Soy Broth with 15% Glycerol in The Deep Freezer (-70 to -80 °C)

For different bacterial preservation techniques, there is no single method applicable for all bacteria. This study aimed to assess the viability of seven species/species groups of clinical bacteria isolates on the long-term storage (more than 5 years) by using Tryptic Soy Broth with 15% glycerol in the deep freezer (-70 to -80°C). A total 10,654 clinical bacteria isolates used as samples in this study. The isolates consisted of seven species/species groups (i.e. Escherichia coli, Campylobacter spp, Shigella spp, Vibrio spp, Salmonella spp, Aeromonas hydrophila, and Neisseria gonorhoeae). The isolates were collected from some previous studies and preserved in the Tryptic Soy Broth (TSB) with 15% glycerol and stored in the deep freezer (-70 to -80°C) for more than five years. The samples were revived on the suitable medium to evaluate the viability of bacteria. Identification conducted by microscopic examination, biochemical test, and latex agglutination. The study showed that the viability of Salmonella spp, Shigella spp, Aeromonas hydrophila and E. coli was 100%, while Campylobacter spp, Vibrio spp, and N. gonorhoeae were 66.7%, 66.4%, and 52.5% respectively. We concluded that viability of Salmonella spp, Shigella spp, A. hydrophila, and E. coli was optimum thus better than Campylobacter spp, Vibrio spp, and N. gonorhoeae for more than 5 years storage by using TSB with 15% glycerol in the deep freezer (-70 to -80 °C).