Antimicrobial resistance and virulence profiling of Staphylococcus pseudintermedius isolated from cats, Bangladesh.

Staphylococcus pseudintermedius is a significant bacterial pathogen that frequently colonizes different body sites and mucous membranes of pets. The objectives of the cross-sectional study were to estimate the prevalence, antimicrobial resistance pattern, and detection of diverse resistance as well as virulence genes of S. pseudintermedius in cats. A standard bacteriological method, species-specific gene and different antimicrobial resistance as well as virulence genes were confirmed by PCR assay. A total of 233 swab samples were collected from different body sites of 102 cats, among them 146 swabs from 73 healthy cats, and 87 from 29 diseased cats. Overall, prevalence of S. pseudintermedius in cats was 12.01%, while dermatitis and otitis affected cats were 26.08% and 33.33%, respectively. The highest antimicrobial resistance was observed against penicillin (96.42%) followed by streptomycin (85.71%) and erythromycin (78.57%). Moreover, 89.28% of S. pseudintermedius isolates exhibit multi-drug resistance (MDR) (≥ 3 classes' antimicrobial resistant). In addition, 17.86% isolates harbored the mecA gene; thus, were classified as methicillin-resistant S. pseudintermedius (MRSP). Furthermore, the erythromycin resistance genes ermA and ermB were harbored by 25% and 10.71% of isolates, while 42.86% and 17.86% of isolates carried tetK and tetL (tetracycline resistance) genes, respectively. In virulence profiling, 32.14% (sea) and 10.71% (seb) of isolates were found positive for enterotoxin genes, whereas, the toxic shock syndrome toxin-1 (tst-1) gene and the Panton-Valentine leukocidin gene (pvl) were detected in 25% and 14.29% of isolates, respectively. To our knowledge, this is the first report of cats in Bangladesh for MDR S. pseudintermedius, MRSP, and their virulence profiling.

Phenotypical Identification and Toxinotyping of Clostridium perfringens Isolates from Healthy and Enteric Disease-Affected Chickens.

Clostridium perfringens is a ubiquitous spore-forming anaerobic pathogen that is frequently associated with enteric disease in chickens. Moreover, enterotoxin-producing C. perfringens has high zoonotic potential as well as serious public health concerns due to the emanation of food-borne intoxication. The present study was designed to isolate, identify, and toxinotype C. perfringens from both healthy and cases of necrotic or ulcerative enteritis chickens. A total of 110 samples were collected from July 2019 to February 2021. Among the samples, 38 (34.5%, 95% CI: 26.39-43.83) were positive for C. perfringens and were obtained from broiler 21 (33.3%, 95% CI: 22.91-45.67), Sonali 9 (34.6%, 95% CI: 19.31-53.88), and layer 8 (38%, 95% CI: 20.68-59.20). C. perfringens was highly prevalent (35.7%, 95% CI: 25.48-47.44) in enteritis chickens compared with healthy ones. In multiplex PCR toxinotyping, 34 (89.4%) isolates were identified as C. perfringens type A by the presence of the alpha toxin gene (cpa). Moreover, in addition to the cpa gene, 3 (14.3%, 95% CI: 4.14-35.48) broiler and 1 (11.1%, 95% CI: 0.01-45.67) Sonali isolates harbored the enterotoxin gene (cpe) and were classified as type F. However, none of the isolates carried genes encoding beta (cpb), epsilon (etx), iota (iap), or beta-2 (cpb2) toxins. Multivariable logistic regression analysis identified the following variables such as; "previously used litter materials" (OR 21.77, 95% CI 2.22-212.66, p ≤ 0.008); intestinal lesions, "presence of ulceration" (OR 30.01, 95% CI 3.02-297.91, p ≤ 0.004); "ballooned with gas" (OR 24.74, 95% CI 4.34-140.86, p ≤ 0.001) and "use of probiotics" (OR 5.24, 95% CI 0.74-36.75, p ≤ 0.095) act as risk factors for C. perfringens colonization in chicken gut. This is the first study of molecular toxinotyping of C. perfringens from healthy and enteric-diseased chickens in Bangladesh, which might have a potential food-borne zoonotic impact on human health.

Severity assessment of single dose Oxford-AstraZeneca vaccinated individuals infected with SARS CoV-2 in the Southeast Bangladesh

The present global endeavor to uncover the most effective vaccines against severe acute respiratory syndrome coronavirus (SARS-CoV-2) that can tremendously prevent transmission, infection and significantly reduce public health risk. COVID-19 vaccination program is underway in different parts of the world including Bangladesh but till to date there is no available health data revealed among the vaccinated peoples. We conducted a cross-sectional study from February 15 to April 15, 2021 to assess the health status of 1st dose Oxford-AstraZeneca vaccinated individuals infected with SARS CoV-2. Standard virological method, real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) was performed to detect SARS-CoV-2 and the different health parameters from vaccinated individuals were collected through direct mobile phone contact using pre-structured questionnaires. A total of 6146 suspected samples were tested and 1752 were found positive for SARS-CoV-2, of them 200 individuals were identified who received 1st dose of COVID-19 vaccine. Within the test period, majority of male (65.6%) and female (34.4%) carried moderate numbers of viruses which comprise between 30.01-35 cyclic threshold (ct) values. Among the vaccinated individuals, 165 (82.5%; 95% CI: 76.51 - 87.5) persons were not hospitalized and 177 (88.5%; 95% CI: 83.24 - 92.57) did not show any respiratory difficulties. Only a few (16) (8%; 95% CI: 4.64 - 12.67) of COVID-19 positive patients needed extra oxygen support and 199 (99.5%; 95% CI: 97.25 - 99.99) individuals didnt require any intensive care unit (ICU) interference. Overall, oxygen saturation was recorded around 96.8% and respiratory difficulties did not extend more than 5 days, irrespective of age and sex during the infection period. Within the vaccinated COVID-19 positive individuals 113 (56.5%; 95% CI: 49.33 - 63.48) and 111(55.5%; 95% CI: 48.32 - 62.51) persons have normal physiological taste and smell. However, we have found a larger proportion of vaccinated persons (129) (64.5%; 95% CI: 57.44 - 71.12) carrying different comorbidity, among them high blood pressure 36 (27.9%; 95% CI, 20.37 - 36.48) and diabetes 32 (24.8%; 95% CI: 17.63 - 33.18) were found more prevalent. Moreover, the significant finding of the present study was 199 (99.5%; 95% CI: 97.25 - 99.99) vaccinated individuals survived with good health conditions and became negative in RT-qPCR. The authors suggest that health risk assessment among the COVID-19 vaccinated persons when infected with SARS-CoV-2 is crucial and time demanding task for the whole world. However, the present study illustrates that the administration of the 1st dose Oxford-AstraZeneca vaccine significantly reduces health risk during the COVID-19 infection period.

Ficus cunia Buch.-Ham. ex Roxb. (leaves): An experimental evaluation of the cytotoxicity, thrombolytic, analgesic and neuropharmacological activities of its methanol extract.

Background The aim of this experiment was to evaluate the cytotoxic, thrombolytic, analgesic, sedative-hypnotic and anxiolytic activities of the methanolic extract of Ficus cunia leaves. Methods Primary phytochemical screening was accomplished by using established methods. Cytotoxicity was studied by brine shrimp lethality test, and the thrombolytic assay was conducted through clot lysis method with human blood. The in vivo action was done using mice of both sexes. The analgesic activity was evaluated by acetic acid-induced writhing test and formalin-induced paw licking test. Open field, hole cross and thiopental Na-induced sleeping time test were used to examine the sedative-hypnotic activity, and elevated plus maze (EPM) and hole board test were used to identify the anxiolytic activity. Results The results elicited that the extract contained several phytochemicals such as alkaloid, flavonoid, and tannin. The extract was found to have a median lethal concentration (LC50) value of 55.48 µg/mL in the brine shrimp lethality bioassay. It was also assessed for antithrombotic activity when compared with streptokinase; it has significant (p < 0.001) thrombolytic effect (34.72 ± 1.74%) contrasted with standard streptokinase (67 ± 1.56%). The extract at doses of 200 and 400 mg/kg produced inhibition of 32.58% and 46.63% in acetic acid-induced pain and 45.88 and 61.18% in formalin-induced pain. The sedative and hypnotic activities on the central nervous system of the methanol extract of F. cunia (MEFC) leaves were evaluated. The extract delivered critical sedative impact at the doses of 200 and 400 mg/kg (by oral route) treated with reference to the substance diazepam, and the hypnotic impact was also observed in the case of mice. MEFC at its maximum dose (400 mg/kg) significantly (p < 0.01) increased the time spent in the open arms of the EPM. In the hole board test, there was a dose-dependent (at 200 and 400 mg/kg) and a significant (p < 0.05 and p < 0.01) increase in the number of head pokes in comparison to control. Conclusions The results of the present study gave a helpful baseline in progression for the possible use of MEFC as a cytotoxic, thrombolytic, analgesic, sedative-hypnotic and anxiolytic drug.

Antinociceptive Activity of Macaranga denticulata Muell. Arg. (Family: Euphorbiaceae): In Vivo and In Silico Studies.

Background: The present study was conducted to investigate the antinociceptive activity of methanol extract of Macaranga denticulata (Met.MD) in an animal model, followed by molecular docking analysis. Methods: Antinociceptive activity was determined by acetic acid-induced writhing and formalin-induced licking test in mice. Then, molecular docking study was performed to identify compounds having maximum activity against the COX-1 enzyme using Schrödinger Maestro (version 10.1) to determine docking fitness. Results: A preliminary phytochemical analysis of Met.MD revealed that it contained alkaloids, carbohydrates, phenols, flavonoids, tannins, and terpenoids. Met.MD exhibited a dose-dependent and statistically significant antinociceptive activity in the acetic acid and formalin test at the doses of 200 and 400 mg/kg. In addition, our docking study showed that macarangin had the best fitness score of -5.81 with COX-1 enzyme among six major compounds of M. denticulata. Conclusions: Results of the present study confirmed the potential antinociceptive activity of M. denticulata leaf extract in both in vivo and in silico models.

Rate of active Helicobacter pylori infection among symptomatic patients of Pakistan

Only few epidemiological studies have examined the rate of active H. pylori infection in the symptomatic population in Pakistan. This retrospective study presents the laboratory data collected during the past 13 years (2002 to 2015) from 2315 symptomatic patients referred to the BreathMAT Lab, Nuclear Medicine, Oncology and Radiotherapy Institute, Islamabad for the diagnosis of active H. pylori infection using the 13C Urea Breath Test. Rate of infection and its association with gender and age were evaluated. The overall rate of active H. pylori infection was 49.5% and there was no association of this rate of infection with gender. An increase in rate of infection was observed with increasing age with significant difference (p < 0.05). The patients that tested negative for this infection might be having symptoms due to stress and indiscriminate use of non-steroidal antiinflammatory drugs (NSAIDs) in this community. The fact that half of the symptomatic patients were negative needs to be highlighted and further suggests that symptomatic patients should be tested by the 13C UBT before prescribing antibiotic treatment for H. pylori eradication. In addition, there is a need to educate this community about the harmful and side effects of self medication and overuse of NSAIDs.

High frequency of cagA and vacA s1a/m2 Genotype among Helicobacter pylori Infected Gastric Biopsies of Pakistani Children

The vacuolating cytotoxin VacA and cytotoxin associated gene product CagA, encoded by vacA and cagA are major virulence determinants associated with pathogenesis of Helicobacter pylori. The presence and prevalence of two major H. pylori virulence associated genes among gastric biopsies of Pakistani children were investigated in the current study. Fifty one gastric biopsy specimens of children were analysed for 16S rRNA, vacA and cagA genes using PCR. The results showed that 21 (41.2%) biopsies were positive for H. pylori as determined by 16S rRNA PCR. In the 21 H. pylori positive gastric biopsies, 19 (90.5%) showed vacA s1a, 1 (4.75%) was vacA s1b and 1 (4.75%) was vacA s2 whereas, 5 (23.8%) were vacA m1 and 16 (76.2%) were vacA m2. None of the H. pylori positive biopsies carried vacA s1c subtype. The cagA gene was found in 13 (61.9%) of H. pylori infected biopsies and different vacA combinations were found with or without cagA gene. H. pylori was detected with high frequency of cagA while vacA s1a and vacA m2 regions with vacA s1a/m2 genotype were predominant in H. pylori infected gastric biopsies of children.