Evaluation of Slit-Skin Smear Against Punch Biopsy in Diagnosing Leprosy: A Cross-sectional Study in a Tertiary Care Centre of West Bengal

Background: Leprosy is an infectious disease which faces diagnostic challenges. Slit-skin smear (SSS) is an age simple diagnostic technique, yet not commonly applied by health care providers. The study aimed to determine the effectiveness of SSS in terms of validity, diagnostic accuracy, and percentage agreement against punch biopsy in diagnosing leprosy among leprosy patients who were diagnosed with leprosy on clinical grounds only (i.e., number of skin lesions and/or peripheral nerve thickening). Methods: An evaluation study of diagnostic tests with a cross-sectional design was conducted at a tertiary care center of Bankura. In general, 70 new untreated leprosy patients, diagnosed solely by clinical grounds (i.e., count of skin lesions and/or thickening of the nerve) and attending the dermatology outpatient department (From February 2019 to January 2020) were enrolled in this study. After excluding pure neuritic, relapse, and seriously ill patients by consecutive sampling, they were subjected to both SSS and punch biopsy using a standard process. SPSS for Windows (Version 16.0., Chicago, SPSS Inc.) was used to analyze data. Z test, chi-square test, and kappa test were conducted to test the statistical significance between the effectiveness of SSS and punch biopsy. Results: The sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratio (DOR), and diagnostic accuracy of SSS were 81.81%, 95.83%, 90%, 92%, 20%, 0.19%, 102.87%, and 91.42%, respectively. Based on the results, SSS could detect acid-fast bacilli (AFB) in clinically diagnosed leprosy cases slightly less than punch biopsy, but it was statistically insignificant (Z=0.3689, P=0.71138, df=1). Finally, Cohen’s Kappa coefficient was 0.796, representing substantial agreement between SSS and punch biopsy (95% CI: 0.641-0.951). Conclusions: Overall, SSS is more or less equally effective as compared to punch biopsy in confirming leprosy cases. Interest and training on SSS among resident doctors should be emphasized accordingly.

A Literature Review of Neospora caninum Infection in Humans

Background: Neosporosis is a parasitic disease caused by Neospora caninum. This parasite is an obligate intracellular coccidia similar to Toxoplasma gondii with a global distribution. With regard to the experimental studies, vertical transmission of the parasite in the monkey (non-human primates) has increased the concern about the zoonotic potential of this disease. The principal aim of the current research was to perform a mini-review on investigations regarding the Neospora infection in humans on a global scale for the first time. Methods: All peer-reviewed articles (published until April 2021) on the Neospora infection in humans were searched in English databases such as Google Scholar, ScienceDirect, Scopus, PubMed, and ProQuest. Results: Based on data in the available articles, the presence of antibodies against the Neospora infection was between 0 and 37.7% in people from different countries. The seroprevalence rate of this infection in HIV-positive individuals was higher (26.6% and 37.7%) compared to other cases. Finally, the genomic DNA of Neospora was detected up to 1% using molecular biology techniques. Conclusions: Overall, the detection of anti-Neospora antibodies in humans indicated that people have been exposed to the parasite. Comprehensive research studies are essential for clarifying the risk factors associated with the Neospora infection in humans. This report provides the baseline information for future researchers. Molecular investigations and genotypic works on N. caninum isolates are highly recommended as well.

Procalcitonin Serum Level in Patients Aged 3-36 Months With Focal Fever Referred to Hospitals in Western Iran in 2020

Background: Diagnosing viral and bacterial infectious diseases in children is of great importance. The conventional treatment for the given diseases has been proven relatively impractical and, therefore, finding a practical diagnostic method seems necessary. Measuring procalcitonin (PCT) levels in the blood is one of those useful tests which have high sensitivity and specificity compared to other methods. Moreover, many researchers have emphasized that the level of PCT in bacterial infections is significant. Therefore, PCT level measurement can be adopted as a highly effective factor for distinguishing bacterial infections from viral ones. Our study aimed to evaluate the plasma levels of PCT in children aged 3-36 months. Methods: In this study which was conducted in 2020 in Kermanshah, Iran, 49 children aged 3-36 months having focal fever and referring to the pediatric emergency department of Mohammad Kermanshahi and Imam Reza hospitals in Kermanshah were examined. Distinguishing bacterial infection from viral one was first made by a pediatrician using CBC diff-ESR-CRP tests. Results: The mean serum level of PCT in bacterial infections was significantly higher than that in viral infections. Furthermore, the mean of white blood cell (WBC), erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP) in bacterial infection was significantly higher than that in viral infection. Conclusions: According to our study findings, plasma levels of PCT could have been considered as a diagnostic indicator of the infection. Therefore, it was recommended that the evaluation of PCT plasma levels in children with infection be performed in early stages of the disease. However, it was also suggested that this evaluation be conducted after performing further investigations in this field.

A Survey of Awareness and Stress of Coronavirus Disease 2019 in a Part of the Iranian Population

Background: The coronavirus disease 2019 (COVID-19) includes a large family of Coronaviridae, which was first found in Wuhan, China in 2019, and has caused a global pandemic. This study aimed was to assess the awareness, stress, and fear of this disease in part of the Iranian population. Methods: Based on the aim of this cross-sectional and analytical study, electronic questionnaires were sent to 201 of the Iranian population. The statistical data from inferential chi-square and Spearman tests were analyzed with IBM SPSS Statistics (version 24), and the level of significance was considered at 0.05. Results: The findings showed that 69.07% of the population was from northern Iran. The awareness level of the majority of people (76.06%) was reported to be favorable. The highest and lowest levels of awareness in people with higher education and unemployed people, respectively, showed meaningful significance between education level and age. Finally, the amount of stress to COVID-19 revealed a significant relationship with age. Conclusions: Based on the obtained results, awareness about age and education was reported to be at the desired level. At the beginning of the outbreak, the anxiety level of people was high while its rate decreased by gaining awareness of the prevention of this debase, indicating the importance of personal health and self-care in society

Application of Multiplex PCR for the Identification of Oxacillinase Genes and Determination of Antibiotic Resistance Pattern in Environmental Isolates of Acinetobacter baumannii in ICU

Background: Acinetobacter baumannii is a common cause of nosocomial infections. A prominent feature of these bacteria is resistance to carbapenems. This study aimed to identify OXA genes encoding oxacillinase in Acinetobacter baumannii isolates. Methods: This cross-sectional descriptive study was performed on 25 environmental A. baumannii isolates collected from ICU over 8 months. Definitive identification of isolates was performed by biochemical tests and polymerase chain reaction (PCR) of 16s rRNA gene. Antibiotic susceptibility testing was performed on Müller-Hinton agar medium by disk diffusion and E-test. Antibiogram and multiplex PCR data of beta-lactamase genes were collected and analyzed at a significance level of P<0.05 using SPSS 22.0. Results: Except for one isolate, all isolates (96%) were sensitive to polymyxin B and 80% of isolates were sensitive to oxacillin. All isolates were sensitive to meropenem, ampicillin/sulbactam, gentamicin, amikacin, piperacillin, and carbenicillin. The results showed that 25 isolates (100%) had OXA-51 gene, 21 isolates (84%) had OXA-58 gene, one isolate (4%) had OXA-24 gene, and none of the isolates contained OXA-23 gene. Only isolate No.10 had three oxacillinase genes simultaneously and it was resistant to oxacillin, polymyxin B, and cephalothin. Conclusions: The study showed that environmental isolates of ICU do not have pathogenic genes present in the clinical isolates, and how these genes are transferred to the peripheral isolates is an important point that should be studied. Identification of genes encoding carbapenem resistance may help to understand the mechanisms of resistance transfer in A. baumannii. The lack of the OXA-23 gene plays an important role in the susceptibility of isolates to antibiotics and non-emergence of resistant strains.

Epidemiological Distribution and Potential Risk Factors of Orientia tsutsugamushi Infection in Eastern Uttar Pradesh, India

Background: Scrub typhus (ST) is a rickettsial infection caused by Orientia tsutsugamushi, which presents with flu like symptoms. This disease has been reported from all over India but with slight variations in its pattern. For decreasing the prevalence, preventing new incidences, and predicting the course of the ST, therefore, it is crucial to gain knowledge and perception of local risk components associated with the disease. The present study aimed to investigate the epidemiological distribution and potential risk factors of O. tsutsugamushi Infection in Eastern Uttar Pradesh (EUP), India. Methods: The serums of 211 samples were collected from the suspected cases along with the detailed information about the participants such as age, location, and place recorded in case history form (CRF). IgM estimation was performed using enzyme-linked immunosorbent assay (ELISA) assay. Results: A total of 58 samples (27.4%) out of 211 ones were found to be positive for IgM antibodies against O. tsutsugamushi bacterium. Furthermore, the results were correlated with epidemiological data such as gender, rural or urban background, pets, and occupation. The results showed that 76.7% of the study participants were from rural areas or had bushes around their houses, 88.3% of them had pets/cattle or frequent encounter with rodents at their houses, and 30.3% of them had no toilet facilities at home. Conclusions: It was concluded that the proximity to pets/cattle, having rodents in closer vicinity, residing in places surrounded by vegetation/farm/bushy areas, and following occupations involving field work increased the chances of getting bitten by mites/chiggers. Overall, Orientia tsutsugamushi prevalence increased in EUP, with respect to clinical features, disease presentation, and laboratory diagnosis can help our community to reduce the mortality caused by this infectious disease.

Antibacterial Activity of New Compounds Based on Nanocomposites

Background: Antimicrobial resistance is a major problem in treatment and public health, and it has been increasing over the last few decades. Hence, serious measurements are needed to overcome this challenge. In this study, we evaluated antibacterial and antifungal activity of some nanocomposites including titanium dioxide (5a), polyimide nanocomposites containing cerium oxide (5b), silver-titanium dioxide nanoparticles prepared under desired conditions (5c), polyaniline/wheat husk ash PANI /WHA (5d), Ag-TiO2 prepared by sol–gel route (5e), and cellulose-graphene (5f) against some bacterial and fungal strains, which are the most common agents in many infectious diseases. Methods: The nanoparticles were prepared in desired condition. The agar dilution and well agar diffusion methods were used for determination of inhibition zoon and minimum inhibitory concentration (MIC) during preliminary evaluation of antimicrobial activity against Staphylococcus aureus, Streptococcus pyogenes, Pseudomonas aeruginosa, Escherichia coli and Candida albicans. Results: The results showed that the nanocomposites had good activity against gram-negative bacteria. Conclusions: Our results showed that the inhibitory activity of these nanocomposites on gram-negative bacteria was better than gram-positive bacteria.

Comparative Analysis of Prevalence and Antibiotic Resistance in Vancomycin-Resistant Enterococcus from Clinical Samples – Demographics and Phenotypes

Background: Of all enterococci species, the most renowned clinically as multidrug-resistant pathogens are Enterococcus faecium and Enterococcus faecalis. Vancomycin-resistant Enterococcus (VRE) species are the principal cause of opportunistic hospital-acquired infections, due to numerous resistance mechanisms. Methods: In this study, the prevalence and antibiotic resistance profiles of VRE according to clinical sources from three selected hospitals in Southwest-Nigeria were investigated. Altogether, 431 samples (urine, rectal, and wound swabs - caesarian section (CS), automobile accidents, and other skin lesions and abrasions) were collected from three selected hospitals in Osun State, Nigeria. Established techniques were employed for the recovery of enterococci and screening for VRE while antibiotic susceptibility tests were carried out by disc diffusion technique. Results: Altogether, 208 (48.3%) enterococci strains were recovered from which 85 (40.9%) were VRE. E. faecium predominated at 71.8% (61/85) and E. faecalis at 28.2% (24/85) as determined by phenotypic characterization. VRE isolates exhibited 100%, 97.6%, and 92.9% resistance to ampicillin, clindamycin, and quinupristin-dalfopristin (Q/D) respectively. The least resistance in-vitro was to tigecycline (27.1%). None of the antibiotics exhibited 100% activity against all the isolates. vanA resistant phenotype was prevalent at 65.9%. E. faecium from all study locations displayed higher levels of resistance than E. faecalis. Multiple antibiotic resistance (MAR) indices in all VRE isolates were ≥0.2, all being multidrug-resistant. Conclusions: The high prevalence rate along with the high level of multidrug resistance observed in the present study is worrisome and poses a continuous threat in the therapy of illnesses triggered by VRE as vancomycin was perceived as a drug of choice to curb enterococcal infections.

The Effect of Bacteriocin Isolated From Lactobacillus rhamnosus on Pseudomonas aeruginosa Lipopolysaccharides

Background: Bacteriocins are heterogeneous inhibitory substances that could affect the bacteria belonging to the same genus. Both gram-positive and gram-negative bacteria produce bacteriocins. One of the best sources of producing bacteriocins is Lactobacillus. The aim of this study was to isolate and purify bacteriocin from Lactobacillus rhamnosus and assess its effects on Pseudomonas aeruginosa and synthesis of its lipopolysaccharide. Methods: L. rhamnosus was prepared and cultured at MRS broth and incubated at 37ºC for 24 hours. Then, the medium was centrifuged for the isolation of bacteriocin and the supernatant was considered as bacteriocin. Antibacterial properties of different concentrations of bacteriocin (50, 100, 200, and 400 μg/mL) against P. aeruginosa were assayed by using agar diffusion and broth micro dilution methods. Also, the effect of bacteriocin against lipopolysaccharide synthesis in P. aeruginosa was analyzed by using one unit of minimum inhibitory concentration (MIC) for bacteriocin. Results: The results showed that all bacteriocin concentrations had antibacterial activity against P. aeruginosa. The MIC value was 31.25 μg/mL and minimal bactericidal concentration (MBC) was 62.5 μg/mL. Also, the synthesis of lipopolysaccharide decreased during P. aeruginosa growth period, and it reached zero after 5 hours. Conclusions: The results of this study showed the antibacterial effect of bacteriocin isolated from L. rhamnosus against P. aeruginosa. In addition, this bacteriocin prevented the lipopolysaccharide synthesis in P. aeruginosa.

Effect of Zinc Oxide Nanoparticles on Loaded Antibiotics Against Multidrug-Resistant Acinetobacter spp.

Background: Metal oxide nanoparticles (NPs) have shown promising efficacy for combating bacterial resistance due to their antibacterial properties. This research investigated the effect of zinc oxide NPs (ZnO-NPs) on the antibacterial activity of conventional antibiotics including ciprofloxacin (CIP), cefotaxime (CTX), and colistin (CST) against multidrug-resistant Acinetobacter isolates. Methods: The disc diffusion method was performed to detect the pattern of antibiotic resistance in isolates. The synthesized ZnO-NPs via the solvothermal method were characterized by field emission scanning electron microscopy (FESEM), X-ray diffraction (XRD), and energy-dispersive X-ray spectroscopy (EDS). Finally, the broth microdilution technique was conducted to demonstrate the antibacterial activity of CIP, CTX, and CST antibiotics with and without a sub-inhibitory concentration of ZnO-NPs. Results: XRD, EDS, and FESEM results confirmed the crystalline structure of ZnO-NPs, and the average size was 100±58.68 nm. All isolates were discovered to be of multidrug-resistant (MDR) type and fully susceptible to CST. The antibacterial activity of CTX and CIP was restored when combined with a sub-inhibitory level of ZnO-NPs (0.25 mg/L), and the highest activity was obtained at the concentrations of 32 µg/mL CTX and 8 µg/ mL CIP. Eventually, ZnO-NPs showed a synergistic effect on the antibacterial properties of CST against MDR Acinetobacter. Conclusions: This research indicated that the combination of ZnO-NPs with some common antibiotics can be considered as a novel strategy for reducing the spread of antibiotic-resistant bacteria.