Search results for: fungal infection

Authors: Preeti Madan, Shalini Malhotra, Nirmaljit Kaur, Charoo Hans, VK Sabarwal

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Hospital is one of the most diverse and complex institutions frequented by people from every walk of life without any distinction between age, sex, gender, religion or intellect. This is over and above the normal inhabitant of hospital i.e. doctors, patients, and paramedical staff. The hospital waste generated 85% is non hazardous, 10% infectious and around 5% are non-infectious but hazardous waste. The management of biomedical waste is still in its infancy. There is a lot of confusion with the problems among the generators, operators, decision makers, and general community about the safe management of biomedical waste prompt action initiated to seek new scientific, safe, and cost-effective management of waste.

Keywords: biomedical waste, nosocomial infection, waste management, hospitals

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Authors: Mengfei Peng, Serajus Salaheen, Debabrata Biswas

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Lactobacillus casei found in the human intestine and mouth is commonly applied for dairy production. Recently, it was found that some byproducts produced by Lactobacillus exhibited antimicrobial activities against multiple bacteria. Meanwhile, introduction of prebiotic-like foods (e.g. cocoa) or probiotics or both of them as food supplements in human diets as well as in farm animal feeds is believed to be an effective ways in control/reduce the colonization of foodborne bacterial pathogens infection in the gut environment. We hypothesized that cocoa may stimulate the production antimicrobial components of Lactobacillus casei and may potentially inhibit/reduce the colonization and infection of foodborne bacterial pathogens in the gut. Mixed culture of L. casei (LC) with enterohemorrhagic E. coli EDL933 (EHEC), Salmonella Typhimurium LT2 (ST), or Listeria monocytogenes LM2 (LM) showed that LC could competitively exclude (100%) them within 72 h. Further, investigation of cell-free culture supernatant (CFCS) revealed that the antimicrobial effects of LC came from CFCS. CFCS of LC eliminated (100%) EHEC, ST, and LM within 72 h, and 2 h CFCS treatment increased the hydrophobicity of EHEC (5.10 folds), ST (8.48 folds), and LM (2.03 folds). In addition, LC cells exhibited more inhibitive effects than CFCS on cell adhesive and invasive activities of EHEC (52.14% & 90.45%), ST (66.89% & 93.83%), and LM (61.10% & 83.40%). Two clusters of poly-peptides in CFCS were identified by SDS-PAGE, the molecular weights of which are ≈5 KD and 40-45 KD. LC CFCS with overnight growth in the presence of 3% strengthened all of the antimicrobial activities (growth inhibition, outer membrane disruption, and cell infective ability reduction). Liquid chromatography/Mass spectrometry analysis detected 5 unique components in class of flavonoids in LC CFCS with overnight 3% cocoa supplement. Furthermore, qPCR results showed that CFCSs up-regulated the expression level of genes responsible for flagellin synthesis and motility, but down-regulated genes for specific binding and invasion-associated proteins synthesis. The stimulatory effects of cocoa in producing bioactive components of probiotics may aid prevention of foodborne illness caused by major foodborne enteric bacterial pathogens.

Keywords: foodborne pathogens, probiotics, prebiotics, pathogen exclusion

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Authors: Nahla Elsherbiny, Ahmed Ahmed, Hamada Mohammed, Mohamed Ali

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Background: The enterococci may be considered as opportunistic agents particularly in immunocompromised patients. It is one of the top three pathogens causing many healthcare associated infections (HAIs). Resistance to several commonly used antimicrobial agents is a remarkable characteristic of most species which may carry various genes contributing to virulence. Objectives: to determine the prevalence of enterococci species in different intensive care units (ICUs) causing health care-associated infections (HAIs), intestinal carriage and environmental contamination. Also, to study the antimicrobial susceptibility pattern of the isolates with special reference to vancomycin resistance. In addition to phenotypic and genotypic detection of gelatinase, cytolysin and biofilm formation among isolates. Patients and Methods: This study was carried out in the infection control laboratory at Assiut University Hospitals over a period of one year. Clinical samples were collected from 285 patients with various (HAIs) acquired after admission to different ICUs. Rectal swabs were taken from 14 cases for detection of enterococci carriage. In addition, 1377 environmental samples were collected from the surroundings of the patients. Identification was done by conventional bacteriological methods and confirmed by analytical profile index (API). Antimicrobial sensitivity testing was performed by Kirby Bauer disc diffusion method and detection of vancomycin resistance was done by agar screen method. For the isolates, phenotypic detection of cytolysin, gelatinase production and detection of biofilm by tube method, Congo red method and microtiter plate. We performed polymerase chain reaction (PCR) for detection of some virulence genes (gelE, cylA, vanA, vanB and esp). Results: Enterococci caused 10.5% of the HAIs. Respiratory tract infection was the predominant type (86.7%). The commonest species were E.gallinarum (36.7%), E.casseliflavus (30%), E.faecalis (30%), and E.durans (3.4 %). Vancomycin resistance was detected in a total of 40% (12/30) of those isolates. The risk factors associated with acquiring vancomycin resistant enterococci (VRE) were immune suppression (P= 0.031) and artificial feeding (P= 0.008). For the rectal swabs, enterococci species were detected in 71.4% of samples with the predominance of E. casseliflavus (50%). Most of the isolates were vancomycin resistant (70%). Out of a total 1377 environmental samples, 577 (42%) samples were contaminated with different microorganisms. Enterococci were detected in 1.7% (10/577) of total contaminated samples, 50% of which were vancomycin resistant. All isolates were resistant to penicillin, ampicillin, oxacillin, ciprofloxacin, amikacin, erythromycin, clindamycin and trimethoprim-sulfamethaxazole. For the remaining antibiotics, variable percentages of resistance were reported. Cytolysin and gelatinase were detected phenotypically in 16% and 48 % of the isolates respectively. The microtiter plate method showed the highest percentages of detection of biofilm among all isolated species (100%). The studied virulence genes gelE, esp, vanA and vanB were detected in 62%, 12%, 2% and 12% respectively, while cylA gene was not detected in any isolates. Conclusions: A significant percentage of enterococci was isolated from patients and environments in the ICUs. Many virulence factors were detected phenotypically and genotypically among isolates. The high percentage of resistance, coupled with the risk of cross transmission to other patients make enterococci infections a significant infection control issue in hospitals.

Keywords: antimicrobial resistance, enterococci, ICUs, virulence factors

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Authors: Ji-Chan Jang

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Tuberculosis is still major health problem because there is an increase of multidrug-resistant and extensively drug-resistant forms of the disease. Therefore, the most urgent clinical need is to discover potent agents and develop novel drug combination capable of reducing the duration of MDR and XDR tuberculosis therapy. Three reference strains H37Rv, CDC1551, W-Beijing GC1237 and six clinical isolates of MDRTB were tested to daptomycin in the range of 0.013 to 256 mg/L. Daptomycin is resistant to all tested M. tuberculosis strains not only laboratory strains but also clinical MDR strains that were isolated at different source. Daptomycin will not be an antibiotic of choice for treating infection of Gram positive atypical slowly growing M. tuberculosis.

Keywords: tuberculosis, daptomycin, resistance, Mycobacterium tuberculosis

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Authors: Carlos E. Rodríguez-Rodríguez, Karla Ruiz-Hidalgo, Kattia Madrigal-Zúñiga, Juan Salvador Chin-Pampillo, Mario Masís-Mora, Elizabeth Carazo-Rojas

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One of the main causes of contamination linked to agricultural activities is the spillage and disposal of pesticides, especially during the loading, mixing or cleaning of agricultural spraying equipment. One improvement in the handling of pesticides is the use of biopurification systems (BPS), simple and cheap degradation devices where the pesticides are biologically degraded at accelerated rates. The biologically active core of BPS is the biomixture, which is constituted by soil pre-exposed to the target pesticide, a lignocellulosic substrate to promote the activity of ligninolitic fungi and a humic component (peat or compost), mixed at a volumetric proportion of 50:25:25. Considering the known ability of lignocellulosic fungi to degrade a wide range of organic pollutants, and the high amount of lignocellulosic waste used in biomixture preparation, the bioaugmentation of biomixtures with these fungi represents an interesting approach for improving biomixtures. The present work aimed at evaluating the effect of the bioaugmentation of rice husk based biomixtures with the fungus Trametes versicolor in the removal of the insectice/nematicide carbofuran (CFN) and to optimize the composition of the biomixture to obtain the best performance in terms of CFN removal and mineralization, reduction in formation of transformation products and decrease in residual toxicity of the matrix. The evaluation of several lignocellulosic residues (rice husk, wood chips, coconut fiber, sugarcane bagasse or newspaper print) revealed the best colonization by T. versicolor in rice husk. Pre-colonized rice husk was then used in the bioaugmentation of biomixtures also containing soil pre-exposed to CFN and either peat (GTS biomixture) or compost (GCS biomixture). After spiking with 10 mg/kg CBF, the efficiency of the biomixture was evaluated through a multi-component approach that included: monitoring of CBF removal and production of CBF transformation products, mineralization of radioisotopically labeled carbofuran (14C-CBF) and changes in the toxicity of the matrix after the treatment (Daphnia magna acute immobilization test). Estimated half-lives of CBF in the biomixtures were 3.4 d and 8.1 d in GTS and GCS, respectively. The transformation products 3-hydroxycarbofuran and 3-ketocarbofuran were detected at the moment of CFN application, however their concentration continuously disappeared. Mineralization of 14C-CFN was also faster in GTS than GCS. The toxicological evaluation showed a complete toxicity removal in the biomixtures after 48 d of treatment. The composition of the GCS biomixture was optimized using a central composite design and response surface methodology. The design variables were the volumetric content of fungally pre-colonized rice husk and the volumetric ratio compost/soil. According to the response models, maximization of CFN removal and mineralization rate, and minimization in the accumulation of transformation products were obtained with an optimized biomixture of composition 30:43:27 (pre-colonized rice husk:compost:soil), which differs from the 50:25:25 composition commonly employed in BPS. Results suggest that fungal bioaugmentation may enhance the performance of biomixtures in CFN removal. Optimization reveals the importance of assessing new biomixture formulations in order to maximize their performance.

Keywords: bioaugmentation, biopurification systems, degradation, fungi, pesticides, toxicity

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Authors: Semira Zeru Haileslassie

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Lack of awareness and practices of PEP treatment were observed among respondents, but they had a better attitude towards PEP. To this end, a formal training for all respondents regarding PEP for HIV prior to their clinical attachments is of utmost importance. The training ought to incorporate a brief clarification with respect to the unpleasant impact of non-adherence that essentially incorporate destitute treatment result and most prominent hazard of resistance and few given as a major cause for non-compliance to PEP, common transient side-effects of PEP and its administrations ought to be cloister educated healthcare specialists to diminish its effect on adherence. Besides, the propensity of detailing needle adhere harm was destitute that needs endeavors to progress. Progressing the culture of detailing and making the detailing handle simple is very necessary. In reality, announcing such wounds as early as conceivable will educate others not to commit same issue once more and, for the most part, will empower stakeholders to intercede the issue sometime prior to it re-occur. At long last, as distant as get up and go utilize has cleared out with so numerous bothers, risk decrease is the foremost choice. With this, taking the increased significance of protective barriers so as to decrease the hazard of exposure to HIV, distinctive stakeholders (the healing center hardware supply chain director, the HIV/ Helps clinic, the clinic chief, hardware and supply quality confirmation group, and other authoritative bodies) ought to work together in co-ordination to secure the supply and guarantee the quality of those crucial protective barriers and to advance demand health laborers to continuously wear protective barriers when exposed to HIV hazard components as well as to dispose appropriately once done. At long last, we prescribe future examiners to conduct planned multicenter studies with extra goals (counting indicator investigation) for way better generalization and result. In spite of satisfactory information and favorable state of mind towards PEP for HIV in most of the respondents, this study uncovered that there were delays in starting, low utilization, and fragmented use of the prescribed PEP. So, health care staff need to progress their practice on PEP of HIV through diverse training program related to PEP of HIV.

Keywords: HIV infection, prophylaxis, knowledge, attitude

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Authors: Alice Huffman, Joseph Hartland, Sam Gibbs

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Dermatology is often a neglected specialty in low-resource settings, despite the high morbidity associated with skin disease. This becomes even more significant when associated with HIV infection, as dermatological conditions are more common and aggressive in HIV positive patients. African countries have the highest HIV infection rates and skin conditions are frequently misdiagnosed and mismanaged, because of a lack of dermatological training and educational material. The frequent lack of diagnostic tests in the African setting renders basic clinical skills all the more vital. This project aimed to improve diagnosis and treatment of skin disease in the HIV population in a district hospital in Malawi. A basic dermatological clinical tool was developed and produced in collaboration with local staff and based on available literature and data collected from clinics. The aim was to improve diagnostic accuracy and provide guidance for the treatment of skin disease in HIV positive patients. A literature search within Embase, Medline and Google scholar was performed and supplemented through data obtained from attending 5 Antiretroviral clinics. From the literature, conditions were selected for inclusion in the resource if they were described as specific, more prevalent, or extensive in the HIV population or have more adverse outcomes if they develop in HIV patients. Resource-appropriate treatment options were decided using Malawian Ministry of Health guidelines and textbooks specific to African dermatology. After the collection of data and discussion with local clinical and pharmacy staff a list of 15 skin conditions was included and a booklet created using the simple layout of a picture, a diagnostic description of the disease and treatment options. Clinical photographs were collected from local clinics (with full consent of the patient) or from the book ‘Common Skin Diseases in Africa’ (permission granted if fully acknowledged and used in a not-for-profit capacity). This tool was evaluated by the local staff, alongside an educational teaching session on skin disease. This project aimed to reduce uncertainty in diagnosis and provide guidance for appropriate treatment in HIV patients by gathering information into one practical and manageable resource. To further this project, we hope to review the effectiveness of the tool in practice.

Keywords: dermatology, HIV, Malawi, skin disease

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Authors: Luciano Tarantino

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Gas gangrene is a rare, severe infection with a very high mortality rate caused by Clostridium species. The infection causes a non-suppurative localized producing gas lesion from which harmful toxins that impair the inflammatory response cause vessel damage and multiple organ failure. Gas gangrene of the liver is very rare and develops suddenly, often as a complication of abdominal surgery and liver transplantation. The present paper deals with a case of gas gangrene of the liver that occurred after percutaneous MW ablation of hepatocellular carcinoma, resulting in progressive liver necrosis and multi-organ failure in spite of specific antibiotics administration. The patient was successfully treated with percutaneous Radiofrequency ablation. Case report: Female, 76 years old, Child A class cirrhosis, treated with synchronous insertion of 3 MW antennae for large HCC (5.5 cm) in the VIII segment. 24 hours after treatment, the patient was asymptomatic and left the hospital . 2 days later, she complained of fever, weakness, abdominal swelling, and pain. Abdominal US detected a 2.3 cm in size gas-containing area, eccentric within the large (7 cm) ablated area. The patient was promptly hospitalized with the diagnosis of anaerobic liver abscess and started antibiotic therapy with Imipenem/cilastatine+metronidazole+teicoplanine. On the fourth day, the patient was moved to the ICU because of dyspnea, congestive heart failure, atrial fibrillation, right pleural effusion, ascites, and renal failure. Blood tests demonstrated severe leukopenia and neutropenia, anemia, increased creatinine and blood nitrogen, high-level FDP, and high INR. Blood cultures were negative. At US, unenhanced CT, and CEUS, a progressive enlargement of the infected liver lesion was observed. Percutaneous drainage was attempted, but only drops of non-suppurative brownish material could be obtained. Pleural and peritoneal drainages gave serosanguineous muddy fluid. The Surgeon and the Anesthesiologist excluded any indication of surgical resection because of the high perioperative mortality risk. Therefore, we asked for the informed consent of the patient and her relatives to treat the gangrenous liver lesion by percutaneous Ablation. Under conscious sedation, percutaneous RFA of GG was performed by double insertion of 3 cool-tip needles (Covidien LDT, USA ) into the infected area. The procedure was well tolerated by the patient. A dramatic improvement in the patient's condition was observed in the subsequent 24 hours and thereafter. Fever and dyspnea disappeared. Normalization of blood tests, including creatinine, was observed within 4 days. Heart performance improved, 10 days after the RFA the patient left the hospital and was followed-up with weekly as an outpatient for 2 months and every two months thereafter. At 18 months follow-up, the patient is well compensated (Child-Pugh class B7), without any peritoneal or pleural effusion and without any HCC recurrence at imaging (US every 3 months, CT every 6 months). Percutaneous RFA could be a valuable therapy of focal GG of the liver in patients non-responder to antibiotics and when surgery and liver transplantation are not feasible. A fast and early indication is needed in case of rapid worsening of patient's conditions.

Keywords: liver tumor ablation, interventional ultrasound, liver infection, gas gangrene, radiofrequency ablation

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Authors: Alketa Qafmolla, Mimoza Canga, Edit Xhajanka, Vergjini Mulo, Ramazan Isufi, Vito Antonio Malagnino

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COVID-19 is threatening the lives of people all over the world. A number of health problems, including oral health problems, have been linked to SARS-CoV-2 infection. Loss of taste is one of the initial symptoms presented by patients who have COVID-19. Purpose: The aim of the current study is to determine the prevalence of taste loss in young adults aged 18 to 26 who have contracted SARS-CoV-2. Materials and methods: This study is analytical cross-sectional research conducted in Albania from March 2023 to September 2023. Our research included a total of 157 students, of which 100 (63.7%) were female and 57 (36.3%) were male. They were divided into three age groups: 18-20, 21-23, and 24-26 years old. Students willingly agreed to participate in the current study and were assured that their participation would be kept anonymous. The study recorded no dropouts and was conducted in accordance with the Declaration of Helsinki. Statistical analysis was performed using IBM SPSS Statistics Version 23.0 on Microsoft Windows Linux, Chicago, IL, USA. The evaluation of data was done using analysis of variance (ANOVA), with a significance level set at P ≤ 0.05. Results: 113 (72%) of the participants reported loss of taste, while 44 (28%) did not experience any loss of taste. According to the study's data analysis, taste problems typically manifest over three days, with the lowest frequency occurring on the second day and the highest frequency occurring on the fifteenth. 68.7% of participants reported experiencing taste recovery after three weeks. The present study's findings demonstrated a substantial correlation between the duration of the individuals' COVID-19 infection and taste loss (P <0.0003). Based on the statistical analysis of the data, this study shows that there is no association between gender and loss of taste (P = 0.218). The participants reported having undergone the following treatments: prednisolone sodium phosphate (15 mg/5 mL daily), vitamin C (1000 mg), azithromycin (500 mg daily), oral vitamin D3 supplementation of 5000 IU daily, vitamin B12 (2.4 mcg daily), zinc 20 mg daily, Augmentin tablets (625 mg), and magnesium sulfate (4 g/100 mL). Conclusion: Within the limitations of this study conducted in Albania, it can be concluded that loss of taste was present in 72% of participants infected with COVID-19 and recovery was evident after three weeks.

Keywords: adult, Albania, COVID-19, cross-sectional study, loss of taste

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Authors: Delsy Salinas, Andreia Garcês, Augusto Silva, Paula Brilhante Simões

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Urinary tract infection (UTI) is a common disease affecting dogs and cats in both community and hospital environment. Bacteria is the most frequent agent isolated, fewer than 1% of infections are due to parasitic, fungal, or viral agents. Common symptoms and laboratory abnormalities includeabdominal pain, pyrexia, renomegaly, and neutrophilia with left shift. A rapid and precise identification of the bacterial agent is still a challenge in veterinarian laboratories. Therefore, this cross-sectional study aims to describe bacterial colony patterns of urine samples by using chromID™ CPS® EliteAgar (BioMérieux, France) from canine and feline specimens submitted to a veterinary laboratory in Portugal (INNO Veterinary Laboratory, Braga)from January to March2022. All urine samples were cultivated in CPS Elite Agar with calibrated 1 µL inoculating loop and incubated at 37ºC for 18-24h. Color,size, and shape (regular or irregular outline)were recorded for all samples. All colonies were classified as Gram-negative or Gram-positive bacteriausing Gram stain (PREVI® Color BioMérieux, France) and determined if they were pure colonies. Identification of bacteria species was performed using GP and GN cards inVitek 2® Compact(BioMérieux, France). A total of 256/1003 submitted urine samples presented bacterial growth, from which 172 isolates were included in this study. The sample’s population included 111 dogs (n=45 males and n=66 females) and 61 cats (n=35 males and n=26 females). The most frequent isolated bacteria wasEscherichia coli (44,7%), followed by Proteus mirabilis (13,4%). All Escherichia coli isolates presented red to burgundy colonies, a colony diameter between 2 to 6 mm, and regular or irregular outlines. Similarly, 100% of Proteus mirabilis isolates were dark yellow colonies with a diffuse pigment and the same size and shape as Escherichia coli. White and pink pale colonies where Staphylococcus species exclusively and S. pseudintermedius was the most frequent (8,2 %). Cian to blue colonies were mostly Enterococcusspp. (8,2%) and Streptococcus spp. (4,6%). Beige to brown colonies were Pseudomonas aeruginosa (2,9%) and Citrobacter spp. (1,2%).Klebsiella spp.,Serratia spp. and Enterobacter spp were green colonies. All Gram-positive isolates were 1 to 2 mm diameter long and had a regular outline, meanwhile, Gram-negative rods presented variable patterns. This results showed that theprevalence of E coli and P. mirabilis as uropathogenic agents follows the same trends in Europe as previously described in other studies. Both agents presented a particular color pattern in CPS Elite Agar to identify them without needing complementary tests. No other bacteria genus could be correlated strongly to a specific color pattern, and similar results have been observed instudies using human’s samples. Chromogenic media shows a great advantage for common urine bacteria isolation than traditional COS, McConkey, and CLEDAgar mediums in a routine context, especially when mixed fermentative Gram-negative agents grow simultaneously. In addition, CPS Elite Agar is versatile for Artificial Intelligent Reading Plates Systems. Routine veterinarian laboratories could use CPS Elite Agar for a rapid screening for bacteria identification,mainlyE coli and P.mirabilis, saving 6h to 10h of automatized identification.

Keywords: cats, CPS elite agar, dogs, urine pathogens

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Authors: Sasiporn Rattanasupha, Settapat Chinviriyasit

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The treatment function adopts a continuous and differentiable function which can describe the effect of delayed treatment when the number of infected individuals increases and the medical condition is limited. In this paper, the SEIR epidemic model with treatment function is studied to investigate the dynamics of the model due to the effect of treatment. It is assumed that the treatment rate is proportional to the number of infective patients. The stability of the model is analyzed. The model is simulated to illustrate the analytical results and to investigate the effects of treatment on the spread of infection.

Keywords: basic reproduction number, local stability, SEIR epidemic model, treatment function

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Authors: Mohd Izzat Md. Nor, Norfazlina Jaffar, Noor Zaitulakma Md. Zain, Nur Izyanti Mohd Suppian, Subhashini Balakrishnan, Geetha Kandavello

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During the Coronavirus (COVID-19) pandemic, Malaysia has been focusing on building herd immunity by introducing vaccination programs into the community. Hospital Standard Operating Procedures (SOP) were developed to prevent inpatient transmission. Objective: In this study, we focus on the positivity rate of inpatient Person Under Surveillance (PUS) becoming COVID-19 positive and compare this to the National rate in order to see the outcomes of the patient who becomes COVID-19 positive in relation to their vaccination status. Methodology: This is a retrospective observational study carried out from 1 January until 30 March 2022 in Institut Jantung Negara (IJN). There were 5,255 patients admitted during the time of this study. Pre-admission Polymerase Chain Reaction (PCR) swab was done for all patients. Patients with positive PCR on pre-admission screening were excluded. The patient who had exposure to COVID-19-positive staff or patients during hospitalization was defined as PUS and were quarantined and monitored for potential COVID-19 infection. Their frequency and risk of exposure (WHO definition) were recorded. A repeat PCR swab was done for PUS patients that have clinical deterioration with or without COVID symptoms and on their last day of quarantine. The severity of COVID-19 infection was defined as category 1-5A. All patients' vaccination status was recorded, and they were divided into three groups: fully immunised, partially immunised, and unvaccinated. We analyzed the positivity rate of PUS patients becoming COVID-positive, outcomes, and correlation with the vaccination status. Result: Total inpatient PUS to patients and staff was 492; only 13 became positive, giving a positivity rate of 2.6%. Eight (62%) had multiple exposures. The majority, 8/13(72.7%), had a high-risk exposure, and the remaining 5 had medium-risk exposure. Four (30.8%) were boostered, 7(53.8%) were fully vaccinated, and 2(15.4%) were partial/unvaccinated. Eight patients were in categories 1-2, whilst 38% were in categories 3-5. Vaccination status did not correlate with COVID-19 Category (P=0.641). One (7.7%) patient died due to COVID-19 complications and sepsis. Conclusion: Within the first quarter of 2022, our institution's positivity rate (2.6%) is significantly lower than the country's (14.4%). High-risk exposure and multiple exposures to positive COVID-19 cases increased the risk of PUS becoming COVID-19 positive despite their underlying vaccination status.

Keywords: COVID-19, boostered, high risk, Malaysia, quarantine, vaccination status

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Authors: A. K. Tursunova, O. V. Chebonenko, A. Zh. Amirkulova, A. O. Abaildayev, O. A. Sapko, Y. M. Dyo, A. Sh. Utarbaeva

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Fusarium solani and its variants cause root and stem rot of plants. Dry rot is the most common disease of potato tubers during storage. The causative agents of fusariosis in contact with plants behave as antagonists, growth stimulants or parasites. The diversity of host-parasite relationships is explained by the parasite’s ability to produce a wide spectrum of biologically active compounds including toxins, enzymes, oligosaccharides, antibiotic substances, enniatins and gibberellins. Many of these metabolites contribute to the creation of compatible relations; others behave as elicitors, inducing various protective responses in plants. An important part of the strategy for developing plant resistance against pathogens is the activation of protein synthesis to produce protective ‘pathogenesis-related’ proteins. The family of PR-proteins known to confer the most protective response is chitinases (EC 3.2.1.14, Cht) and β-1,3-glucanases (EC 3.2.1.39, Glu). PR-proteins also include a large multigene family of peroxidases (EC 1.11.1.7, Pod), and increased activity of Pod and expression of the Pod genes leads to the development of resistance to a broad class of pathogens. Despite intensive research on the role of PR-proteins, the question of their participation in the mechanisms of formation of the F.solani–S.tuberosum pathosуstem is not sufficiently studied. Our aim was to investigate the effect of different classes of F. solani metabolites on the activity of chitinase, β-1,3-glucanases and peroxidases in tubers of Solanum tuberosum. Metabolite culture filtrate (CF) and cytoplasmic components were fractionated by extraction of the mycelium with organic solvents, salting out techniques, dialysis, column chromatography and ultrafiltration. Protein, lipid, carbohydrate and polyphenolic fractions of fungal metabolites were derived. Using enzymatic hydrolysis we obtained oligo glycans from fungal cell walls with different molecular weights. The activity of the metabolites was tested using potato tuber discs (d = 16mm, h = 5mm). The activity of PR-proteins of tubers was analyzed in a time course of 2–24 hours. The involvement of the analysed metabolites in the modulation of both early non-specific and late related to pathogenesis reactions was demonstrated. The most effective inducer was isolated from the CF (fraction of total phenolic compounds including naphtazarins). Induction of PR-activity by this fraction was: chitinase - 340-360%, glucanase - 435-450%, soluble forms of peroxidase - 400-560%, related forms of peroxidase - 215-237%. High-inducing activity was observed by the chloroform and acetonitrile extracts of the mycelium (induction of chitinase and glucanase activity was 176-240%, of soluble and bound forms of peroxidase - 190-400%). The fraction of oligo glycans mycelium cell walls of 1.2 kDa induced chitinase and β-1,3-glucanase to 239-320%; soluble forms and related peroxidase to 198-426%. Oligo glycans cell walls of 5-10 kDa had a weak suppressor effect - chitinase (21-25%) and glucanase (25-28%) activity; had no effect on soluble forms of peroxidase, but induced to 250-270% activity related forms. The CF polysaccharides of 8.5 kDa and 3.1 kDa inhibited synchronously the glucanase and chitinase specific response in step (after 24 hours at 42-50%) and the step response induced nonspecific peroxidase activity: soluble forms 4.8 -5.2 times, associated forms 1.4-1.6 times.

Keywords: fusarium solani, PR-proteins, peroxidase, solanum tuberosum

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Authors: A. A. Agbor, Jean Joel R. Bigna, Serges Clotaire Billong, Mathurin Cyrille Tejiokem, Gabriel L. Ekali, Claudia S. Plottel, Jean Jacques N. Noubiap, Hortence Abessolo, Roselyne Toby, Sinata Koulla-Shiro

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Background: Contributors to fatal outcomes in patients undergoing tuberculosis (TB) treatment in the setting of HIV co-infection are poorly characterized, especially in sub-Saharan Africa. Our study’s aim was to assess factors associated with death in TB/HIV co-infected patients during the first 6 months their TB treatment. Methods: We conducted a tertiary-care hospital-based retrospective cohort study from January 2006 to December 2013 at the Yaoundé Central Hospital, Cameroon. We reviewed medical records to identify hospitalized co-infected TB/HIV patients aged 15 years and older. Death was defined as any death occurring during TB treatment, as per the World Health Organization’s recommendations. Logistic regression analysis identified factors associated with death. Magnitudes of associations were expressed by adjusted odds ratio (aOR) with 95% confidence interval. A p value < 0.05 was considered statistically significant. Results: The 337 patients enrolled had a mean age of 39.3 (+/- 10.3) years and more (54.3%) were women. TB treatment outcomes included: treatment success in 60.8% (n=205), death in 29.4% (n=99), not evaluated in 5.3% (n=18), loss to follow-up in 5.3% (n=14), and failure in 0.3% (n=1) . After exclusion of patients lost to follow-up and not evaluated, death in TB/HIV co-infected patients during TB treatment was associated with: a TB diagnosis made before national implementation of guidelines regarding initiation of antiretroviral therapy (aOR = 2.50 [1.31-4.78]; p = 0.006), the presence of other AIDS-defining infections (aOR = 2.73 [1.27-5.86]; p = 0.010), non-AIDS comorbidities (aOR = 3.35 [1.37-8.21]; p = 0.008), not receiving co-trimoxazole prophylaxis (aOR = 3.61 [1.71-7.63]; p = 0.001), not receiving antiretroviral therapy (aOR = 2.45 [1.18-5.08]; p = 0.016), and CD4 cell counts < 50 cells/mm3 (aOR = 16.43 [1.05-258.04]; p = 0.047). Conclusions: The success rate of anti-tuberculosis treatment among hospitalized TB/HIV co-infected patients in our setting is low. Mortality in the first 6 months of treatment was high and strongly associated with specific clinical factors including states of greater immunosuppression, highlighting the urgent need for targeted interventions, including provision of anti-retroviral therapy and co-trimoxazole prophylaxis in order to enhance patient outcomes.

Keywords: TB/HIV co-infection, death, treatment outcomes, factors

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Authors: Sami Michael Ayodele

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Cordyceps are members of the fungi family Cordycepitaceae which are formally in Clavicipitaceae. They parasitize the larvae and adults of different susceptible arthropods. They are highly priced and have been reported to be highly medicinal. Cordyceps have been used for centuries in traditional medicine in China and other Asian countries. Survey studies were carried out to determine the presence of Cordyceps and another entomopathogenic fungal floral in North Central Nigeria for two consecutive years. Thirty samples were collected, twenty-eight were identified, and two were unidentified. Most identified species belong to Ophiocordyceps species (20samples), Cordyceps species (4samples) and Isaria species (4samples). The species identified were: Cordyceps pseudomilitaris, Cordyceps tuberculata, Cordyceps cylindrical, Ophiocordyceps nutans, O. criminals, O. oxycephala, O. kniphofioides and Isaria sinclairii. The morphological and microscopic features of the collected and identified species were similar to those reported in other countries. This is the first official report on the presence of Cordyceps species in Nigeria. Further collections from different ecological regions of Nigeria will show the richness of the floral diversity of these entomopathogenic and medicinal mushrooms in Nigeria.

Keywords: cordyceps, entomopathogenic, medicinal, North Central, Nigeria

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Authors: Swati Gautam, Salman Akhtar, S. P. Jaiswar, Amita Jain

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Background: FGTB is now a major global health problem mostly in developing countries including India. In humans, Mycobacterium Tuberculosis (M.tb) is a causating agent of infection. High index of suspicion is required for early diagnosis due to asymptomatic presentation of FGTB disease. In macrophages Toll Like Receptor-2 (TLR-2) is one which mediated host’s immune response to M.tb. The expression of TLR-2 on macrophages is important to determine the fate of innate immune responses to M.tb. TLR-2 have two work. First its high expression on macrophages worsen the outer of infection and another side, it maintains M.tb to its dormant stage avoids activation of M.tb from latent phase. Single Nucleotide Polymorphism (SNP) of TLR-2 gene plays an important role in susceptibility to TB among different populations and subsequently, in the development of infertility. Methodology: This Case-Control study was done in the Department of Obs and Gynae and Department of Microbiology at King George’s Medical University, U.P, Lucknow, India. Total 300 subjects (150 Cases and 150 Controls) were enrolled in the study. All subjects were enrolled only after fulfilling the given inclusion and exclusion criteria. Inclusion criteria: Age 20-35 years, menstrual-irregularities, positive on Acid-Fast Bacilli (AFB), TB-PCR, (LJ/MGIT) culture in Endometrial Aspiration (EA). Exclusion criteria: Koch’s active, on ATT, PCOS, and Endometriosis fibroid women, positive on Gonococal and Chlamydia. Blood samples were collected in EDTA tubes from cases and healthy control women (HCW) and genomic DNA extraction was carried out by salting-out method. Genotyping of TLR2 genetic variants (Arg753Gln and Arg677Trp) were performed by using single amplification refractory mutation system (ARMS) PCR technique. PCR products were analyzed by electrophoresis on 1.2% agarose gel and visualized by gel-doc. Statistical analysis of the data was performed using the SPSS 16.3 software and computing odds ratio (OR) with 95% CI. Linkage Disequiliribium (LD) analysis was done by SNP stats online software. Results: In TLR-2 (Arg753Gln) polymorphism significant risk of FGTB observed with GG homozygous mutant genotype (OR=13, CI=0.71-237.7, p=0.05), AG heterozygous mutant genotype (OR=13.7, CI=0.76-248.06, p=0.03) however, G allele (OR=1.09, CI=0.78-1.52, p=0.67) individually was not associated with FGTB. In TLR-2 (Arg677Trp) polymorphism a significant risk of FGTB observed with TT homozygous mutant genotype (OR= 0.020, CI=0.001-0.341, p < 0.001), CT heterozygous mutant genotype (OR=0.53, CI=0.33-0.86, p=0.014) and T allele (OR=0.463, CI=0.32-0.66, p < 0.001). TT mutant genotype was only found in FGTB cases and frequency of CT heterozygous more in control group as compared to FGTB group. So, CT genotype worked as protective mutation for FGTB susceptibility group. In haplotype analysis of TLR-2 genetic variants, four possible combinations, i.e. (G-T, A-C, G-C, and A-T) were obtained. The frequency of haplotype A-C was significantly higher in FGTB cases (0.32). Control group did not show A-C haplotype and only found in FGTB cases. Conclusion: In conclusion, study showed a significant association with both genetic variants of TLR-2 of FGTB disease. Moreover, the presence of specific associated genotype/alleles suggest the possibility of disease severity and clinical approach aimed to prevent extensive damage by disease and also helpful for early detection of disease.

Keywords: ARMS, EDTA, FGTB, TLR

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Authors: Daria Reczynska, Justyna Jarczak, Michal Czopowicz, Danuta Sloniewska, Karina Horbanczuk, Wieslaw Jarmuz, Jaroslaw Kaba, Emilia Bagnicka

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Since people, animals and plants are constantly exposed to pathogens they have developed very complex systems of defense. Among ca. 1000 antimicrobial peptides from different families so far identified, approximately 30 belonging to cathelicidin family can be found in mammals. Cathelicidins probably constitute the first line of defense because they can act at a physiological salt concentration which is present in healthy tissues. Moreover, the low salt concentration which is present in infected tissues inhibits their activity. In goat bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), myeloid antimicrobial peptide 28 (MAP28), myeloid antimicrobial peptide 34 (MAP34 A and B), goat bactenecin3.4 (ChBac3.4) were identified. Caprine arthritis-encephalitis (CAE) caused by small ruminant lentivirus (SRLV) is economic problem. The main CAE symptoms are weight loss, arthritis, pneumonia and mastitis (significant elevation of the somatic cell count and deterioration of some technological parameters). The study was conducted on 24 dairy goats. The animals were divided into two groups: experimental (SRLV-infected) and control (non-infected). The blood samples were collected five times: on the 1st, 7th, 30th, 90th and 150thday of lactation. The levels of transcripts of BAC7.5, BAC5, MAP28 and MAP34 genes in blood leucocytes were measured using qPCR method. There were no differences in mRNA levels of studied genes between stages of lactation. The differences were observed in expressions of BAC5, MAP28 and MAP34 genes with lower levels in the experimental group. There was no difference in BAC7.5 expression between groups. The decreased levels of transcripts of cathelicidin genes in blood leucocytes of SRLV-infected goats may indicate the disturbances of homeostasis in organisms. It can be concluded that SRLV infection seems to inhibit expression of cathelicidin genes. The study was financed by a grant from the National Scientific Center No. UMO-2013/09/B/NZ/03514.

Keywords: goat, CAEV, cathelicidins, blood leukocytes, gene expression

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Authors: Basma Hassabo, Sarah Ahmed, Aisha Hameed

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Aims and Objectives: To determine the incidence of maternal mortality amongst pregnant women with sickle cell disease (SCD) in the United Kingdom and to determine exact cause of death in these women. Background: SCD is caused by the ‘sickle’ gene and is characterized by episodes of severe bone pain and other complications like acute chest syndrome, chronic pulmonary hypertension, stroke, retinopathy, chronic renal failure, hepato-splenic crises, avascular bone necrosis, sepsis and leg ulcers. SCD is a continual cause of maternal mortality and fetal complications, and it comprises 1.5% of all Direct and Indirect deaths in the UK. Sepsis following premature rupture of membranes with ascending infection, post-partum infection and pre-labour overwhelming septic shock is one of its leading causes of death. Over the last fifty years of maternal mortality reports in UK, between 1 to 4 pregnant women died in each triennium. Material and Method: This is a retrospective study that involves pregnant women who died from SCD complications in the UK between 1952-2012. Data were collected from the UK Confidential Enquiries into Maternal Death and its causes between 1952–2012. Prior to 1985, exact cause of death in this cohort was not recorded. Results: 33 deaths reported between 1964 and 1984. 17 deaths were reported due to sickle cell disease between 1985 and 2012. Five women in this group died of sickle cell crisis, one woman had liver sequestration crisis, two women died of venous thromboembolism, two had myocardial fibrosis and three died of sepsis. Remaining women died of amniotic fluid embolism, SUDEP, myocardial ischemia and intracranial haemorrhage. Conclusion: The leading causes of death in sickle cell sick pregnant women are sickle cell crises, sepsis, venous thrombosis and thromboembolism. Prenatal care for women with SCD should be managed by a multidisciplinary team that includes an obstetrician, nutritionist, primary care physician, and haematologist. In every sick Sickle Cell woman Sickle Cell crises should be on the top of the list of differential diagnosis. Aggressive treatment of complications with low threshold to commence broad-spectrum antibiotics and LMWH contribute to better outcomes.

Keywords: incidence, maternal mortality, sickle cell disease (SCD), uk

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Authors: Sumit Kanwar, Manisha Chand, Gregory Gilot

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Background: Septic arthritis of the shoulder has infrequently been discussed. Focus on infection of the untouched shoulder has not heretofore been described. We present four patients with glenohumeral septic arthritis. Methods: Case 1: A 59 year old male with left shoulder pain in the anterior, posterior and superior aspects. Case 2: A 60 year old male with fever, chills, and generalized muscle aches. Case 3: A 70 year old male with right shoulder pain about the anterior and posterior aspects. Case 4: A 55 year old male with global right shoulder pain, swelling, and limited ROM. Results: In case 1, the left shoulder was affected. Physical examination, swelling was notable, there was global tenderness with a painful range of motion (ROM). The lab values indicated an erythrocyte sedimentation rate (ESR) of 96, and a C-reactive protein (CRP) of 304.30. Imaging studies were performed and MRI indicated a high suspicion for an abscess with osteomyelitis of the humeral head. Our second case’s left arm was affected. He had swelling, global tenderness and painful ROM. His ESR was 38, CRP was 14.9. X-ray showed severe arthritis. Case 3 differed with the right arm being affected. Again, global tenderness and painful ROM was observed. His ESR was 94, and CRP was 10.6. X-ray displayed an eroded glenoid space. Our fourth case’s right shoulder was affected. He had global tenderness and painful, limited ROM. ESR was 108 and CRP was 2.4. X-ray was non-significant. Discussion: Monoarticular septic arthritis of the virgin glenohumeral joint is seldom diagnosed in clinical practice. Common denominators include elevated ESR, painful, limited ROM, and involvement of the dominant arm. The male population is more frequently affected with an average age of 57. Septic arthritis is managed with incision and drainage or needle aspiration of synovial fluid supplemented with 3-6 weeks of intravenous antibiotics. Due to better irrigation and joint visualization, arthroscopy is preferred. Open surgical drainage may be indicated if the above methods fail. Conclusion: If a middle-aged male presents with vague anterior or posterior shoulder pain, elevated inflammatory markers and a low grade fever, an x-ray should be performed. If this displays degenerative joint disease, the complete further workup with advanced imaging, such as an MRI, CT scan, or an ultrasound. If these imaging modalities display anterior space joint effusion with soft tissue involvement, we can suspect septic arthritis of the untouched glenohumeral joint and surgery is indicated.

Keywords: glenohumeral joint, identification, infection, septic arthritis, shoulder

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Authors: Noura Kichou, Manel Tafergguenit, Nabila Ghechtouli, Zakia Hank

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The aim of this work is to synthesize, characterize and evaluate the biological activity of two Ligands : glyoxime and dimethylglyoxime, and their metal Ni(II) chelates. The newly chelates were characterized by elemental analysis, IR, EPR, nuclear magnetic resonances (1H and 13C), and biological activity. The antibacterial and antifungal activities of the ligands and its metal complexes were screened against bacterial species (Staphylococcus aureus, Bacillus subtilis, and Escherichia coli) and fungi (Candida albicans). Ampicillin and amphotericin were used as references for antibacterial and antifungal studies. The activity data show that the metal complexes have a promising biological activity comparable with parent free ligand against bacterial and fungal species. A structural, energetic, and electronic theoretical study was carried out using the DFT method, with the functional B3LYP and the gaussian program 09. A complete optimization of geometries was made, followed by a calculation of the frequencies of the normal modes of vibration. The UV spectrum was also interpreted. The theoretical results were compared with the experimental data.

Keywords: glyoxime, dimetylglyoxime, nickel, antibacterial activity

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Authors: Noura Kichou, Manel Tafergguenit, Nabila Ghechtouli, Zakia Hank

Abstract:

The aim of this work is to synthesize, characterize and evaluate the biological activity of two Ligands: glyoxime and dimethylglyoxime, and their metal Ni(II) chelates. The newly chelates were characterized by elemental analysis, IR, EPR, nuclear magnetic resonances (1H and 13C), and biological activity. The antibacterial and antifungal activities of the ligands and its metal complexes were screened against bacterial species (Staphylococcus aureus, Bacillus subtilis, and Escherichia coli) and fungi (Candida albicans). Ampicillin and amphotericin were used as references for antibacterial and antifungal studies. The activity data show that the metal complexes have a promising biological activity comparable with parent free ligand against bacterial and fungal species. A structural, energetic, and electronic theoretical study was carried out using the DFT method, with the functional B3LYP and the gaussian program 09. A complete optimization of geometries was made, followed by a calculation of the frequencies of the normal modes of vibration. The UV spectrum was also interpreted. The theoretical results were compared with the experimental data.

Keywords: glyoxime, dimetylglyoxime, nickel, antibacterial activity

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Authors: Getahun Nigusie Demise

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Background; antiretroviral treatment reduce HIV-related morbidity, and prolonged survival of patients however, there is lack of up-to-date information concerning the treatment long term effect on the survival of HIV positive children especially in the study area. Objective: The aim of this study is to assess the incidence and predictors of mortality among HIV positive children on antiretroviral therapy (ART) in public hospitals of Harer town who were enrolled from 2011 to 2021. Methodology: Institution based retrospective cohort study was conducted among 429 HIV positive children enrolled in ART clinic from January 1st 2011 to December30th 2021. Data were collected from medical cards by using a data extraction form, Descriptive analyses were used to Summarized the results, and life table was used to estimate survival probability at specific point of time after introduction of ART. Kaplan Meier survival curve together with log rank test was used to compare survival between different categories of covariates, and Multivariate Cox-proportional hazard regression model was used to estimate adjusted Hazard rate. Variables with p-values ≤0.25 in bivariable analysis were candidates to the multivariable analysis. Finally, variables with p-values < 0.05 were considered as significant variables. Results: The study participants had followed for a total of 2549.6 child-years (30596 child months) with an overall mortality rate of 1.5 (95% CI: 1.1, 2.04) per 100 child-years. Their median survival time was 112 months (95% CI: 101–117). There were 38 children with unknown outcome, 39 deaths, and 55 children transfer out to different facility. The overall survival at 6, 12, 24, 48 months were 98%, 96%, 95%, 94% respectively. being in WHO clinical Stage four (AHR=4.55, 95% CI:1.36, 15.24), having anemia(AHR=2.56, 95% CI:1.11, 5.93), baseline low absolute CD4 count (AHR=2.95, 95% CI: 1.22, 7.12), stunting (AHR=4.1, 95% CI: 1.11, 15.42), wasting (AHR=4.93, 95% CI: 1.31, 18.76), poor adherence to treatment (AHR=3.37, 95% CI: 1.25, 9.11), having TB infection at enrollment (AHR=3.26, 95% CI: 1.25, 8.49),and no history of change their regimen(AHR=7.1, 95% CI: 2.74, 18.24), were independent predictors of death. Conclusion: more than half of death occurs within 2 years. Prevalent tuberculosis, anemia, wasting, and stunting nutritional status, socioeconomic factors, and baseline opportunistic infection were independent predictors of death. Increasing early screening and managing those predictors are required.

Keywords: human immunodeficiency virus-positive children, anti-retroviral therapy, survival, treatment, Ethiopia

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Authors: Amr Saeb, Khalid Al-Rubeaan, Mohamed Abouelhoda, Manojkumar Selvaraju, Hamsa Tayeb

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Background: P. mirabilis is a common uropathogenic bacterium that can cause major complications in patients with long-standing indwelling catheters or patients with urinary tract anomalies. In addition, P. mirabilis is a common cause of chronic osteomyelitis in diabetic foot ulcer (DFU) patients. Methodology: P. mirabilis SCDR1 was isolated from a diabetic ulcer patient. We examined P. mirabilis SCDR1 levels of resistance against nano-silver colloids, the commercial nano-silver and silver containing bandages and commonly used antibiotics. We utilized next generation sequencing techniques (NGS), bioinformatics, phylogenetic analysis and pathogenomics in the identification and characterization of the infectious pathogen. Results: P. mirabilis SCDR1 is a multi-drug resistant isolate that also showed high levels of resistance against nano-silver colloids, nano-silver chitosan composite and the commercially available nano-silver and silver bandages. The P. mirabilis-SCDR1 genome size is 3,815,621 bp with G+C content of 38.44%. P. mirabilis-SCDR1 genome contains a total of 3,533 genes, 3,414 coding DNA sequence genes, 11, 10, 18 rRNAs (5S, 16S, and 23S), and 76 tRNAs. Our isolate contains all the required pathogenicity and virulence factors to establish a successful infection. P. mirabilis SCDR1 isolate is a potential virulent pathogen that despite its original isolation site, wound, it can establish kidney infection and its associated complications. P. mirabilis SCDR1 contains several mechanisms for antibiotics and metals resistance including, biofilm formation, swarming mobility, efflux systems, and enzymatic detoxification. Conclusion: P. mirabilis SCDR1 is the spontaneous nano-silver resistant bacterial strain. P. mirabilis SCDR1 strain contains all reported pathogenic and virulence factors characteristic for the species. In addition, it possesses several mechanisms that may lead to the observed nano-silver resistance.

Keywords: Proteus mirabilis, multi-drug resistance, silver nanoparticles, resistance, next generation sequencing techniques, genome analysis, bioinformatics, phylogeny, pathogenomics, diabetic foot ulcer, xenobiotics, multidrug resistance efflux, biofilm formation, swarming mobility, resistome, glutathione S-transferase, copper/silver efflux system, altruism

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Authors: Sweta Pandey, Samridhi Dhyani, Susmita Chaudhuri

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Klebsiella pneumoniae bacteria is a global threat to human health due to an increase in multi-drug resistance among strains. The hypervirulent strains of Klebsiella pneumoniae is a major trouble due to their association with life-threatening infections in a healthy population. One of the major virulence factors of hyper virulent strains of Klebsiella pneumoniae is the T6SS (Type six secretary system) which is majorly involved in microbial antagonism and causes interaction with the host eukaryotic cells during infections. T6SS mediates some of the crucial factors for establishing infection by the bacteria, such as cell adherence, invasion, and subsequent in vivo colonisation. The antibacterial activity and the cell invasion property of the T6SS system is a major requirement for the establishment of K. pneumoniae infections within the gut. The T6SS can be an appropriate target for developing therapeutics. The T6SS consists of an inner tube comprising hexamers of Hcp (Haemolysin -regulated protein) protein, and at the top of this tube sits VgrG (Valine glycine repeat protein G); the tip of the machinery consists of PAAR domain containing proteins which act as a delivery system for bacterial effectors. For this study, immune response to recombinant VgrG protein was generated to establish this protein as a potential immunogen for the development of therapeutic leads. The immunogenicity of the selected protein was determined by predicting the B cell epitopes by the BCEP analysis tool. The gene sequence for multiple domains of VgrG protein (phage_base_V, T6SS_Vgr, DUF2345) was selected and cloned in pMAL vector in E. coli. The construct was subcloned and expressed as a fusion protein of 203 residue protein with mannose binding protein tag (MBP) to enhance solubility and purification of this protein. The purified recombinant VgrG fusion protein was used for mice immunisation. The antiserum showed reactivity with the recombinant VgrG in ELISA and western blot. The immunised mice were challenged with K. pneumoniae bacteria and showed bacterial clearance in immunised mice. The recombinant VgrG protein can further be used for studying downstream signalling of VgrG protein in mice during infection and for therapeutic MAb development to eradicate K. pneumoniae infections.

Keywords: immune response, Klebsiella pneumoniae, multi-drug resistance, recombinant protein expression, T6SS, VgrG

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Authors: A. Díaz-Roa, P. I. Silva Junior, F. J. Bello

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Sarconesiopsis magellanica (Diptera: Calliphoridae) is a medically important necrophagous fly which is used for establishing the post-mortem interval. Dipterous maggots release diverse proteins and peptides contained in larval excretion and secretion (ES) products playing a key role in digestion. The most important mechanism for combating infection using larval therapy depends on larval ES. These larvae are protected against infection by a diverse spectrum of antimicrobial peptides (AMPs), one already known like lucifensin. Special interest in these peptides has also been aroused regarding understanding their role in wound healing since they degrade necrotic tissue and kill different bacteria during larval therapy. The action of larvae on wounds occurs through 3 mechanisms of action: removal of necrotic tissue, stimulation of granulation tissue, and antibacterial action of larval ES. Some components of the ES include calcium, urea, allantoin ammonium bicarbonate and reducing the viability of Gram positive and Gram negative bacteria. The Lucilia sericata fly larvae have been the most used, however, we need to evaluate new species that could potentially be similar or more effective than fly above. This study was thus aimed at identifying and characterizing S. magellanica AMPs contained in ES products for the first time and compared them with the common fly used L. sericata. These products were obtained from third-instar larvae taken from a previously established colony. For the first analysis, ES fractions were separate by Sep-Pak C18 disposable columns (first step). The material obtained was fractionated by RP-HPLC by using Júpiter C18 semi-preparative column. The products were then lyophilized and their antimicrobial activity was characterized by incubation with different bacterial strains. The first chromatographic analysis of ES from L. sericata gives 6 fractions with antimicrobial activity against Gram-positive bacteria Micrococus luteus, and 3 fractions with activity against Gram-negative bacteria Pseudomonae aeruginosa while the one from S. magellanica gaves 1 fraction against M. luteus and 4 against P. aeruginosa. Maybe one of these fractions could correspond to the peptide already known from L. sericata. These results show the first work for supporting further experiments aimed at validating S. magellanica use in larval therapy. We still need to search if we find some new molecules, by making mass spectrometry and ‘de novo sequencing’. Further studies are necessary to identify and characterize them to better understand their functioning.

Keywords: antimicrobial peptides, larval therapy, Lucilia sericata, Sarconesiopsis magellanica

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Authors: Norbert W. Paul

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In 1988, parts of the scientific community still heralded findings to support that AIDS was likely to remain largely a ‘gay disease’. The value-ladden terminology of some of the articles suggested that rectum and fragile urethra are not sufficiently robust to provide a barrier against infectious fluids, especially body fluids contaminated with HIV while the female vagina, would provide natural protection against injuries and trauma facilitating HIV-infection. Anal sexual intercourse was constituted not only as dangerous but also as unnatural practice, while penile-vaginal intercourse would follow natural design and thus be relatively safe practice minimizing the risk of HIV. Statements like the latter were not uncommon in the early times of HIV/AIDS and contributed to captious certainties and an underestimation of heterosexual risks. Pseudo-scientific discourses on the origin of HIV were linked to local and global health politics in the 1980ies. The pathways of infection were related to normative concepts like deviant, subcultural behavior, cultural otherness, and guilt used to target, tag and separate specific groups at risk from the ‘normal’ population. Controlling populations at risk became the top item on the agenda rather than controlling modes of transmission and the virus. Hence, the Thai strategy to cope with HIV/AIDS by acknowledging social and sexual practices as they were – not as they were imagined – has become a role model for successful prevention in the highly scandalized realm of sexually transmitted disease. By accepting the globalized character of local HIV-risk and projecting the risk onto populations which are neither particularly vocal groups nor vested with the means to strive for health and justice Thailand managed to culturally implement knowledge-based tools of prevention. This paper argues, that pertinent cultural collisions regarding our strategies to cope with HIV/AIDS are deeply rooted in misconceptions, misreadings and scandalizations brought about in the early history of HIV in the 1980ties. The Thai strategy is used to demonstrate how local values can be balanced against globalized health risk and used to effectuated prevention by which knowledge and norms are translated into local practices. Issues of global health and injustice will be addressed in the final part of the paper dealing with the achievability of health as a human right.

Keywords: bioethics, HIV, global health, justice

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Authors: Mario Tauzene Afonso Matangue, Ivan Andres Sanchez Ortiz

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The Metropolitan area of Maputo (Mozambique Capital City) is located in semi-arid zone (800 mm annual rainfall) with 1101170 million inhabitants. On the west side, there are the flatlands of Infulene where the Mulauze River flows towards to the Indian Ocean, receiving at this site, the storm water contaminated with sanitary sewage from Maputo, transported through a concrete open channel. In Infulene, local communities grow salads crops such as tomato, onion, garlic, lettuce, and cabbage, which are then commercialized and consumed in several markets in Maputo City. Lettuce is the most daily consumed salad crop in different meals, generally in fast-foods, breakfasts, lunches, and dinners. However, the risk of infection by several pathogens due to the consumption of lettuce, using the Quantitative Microbial Risk Assessment (QMRA) tools, is still unknown since there are few studies or publications concerning to this matter in Mozambique. This work is aimed at determining the annual risk arising from the consumption of lettuce grown in Infulene valley, in Maputo, using QMRA tools. The exposure model was constructed upon the volume of contaminated water remaining in the lettuce leaves, the empirical relations between the number of pathogens and the indicator of microorganisms (E. coli), the consumption of lettuce (g) and reduction of pathogens (days). The reference pathogens were Vibrio cholerae, Cryptosporidium, norovirus, and Ascaris. The water quality samples (E. coli) were collected in the storm water channel from January 2016 to December 2018, comprising 65 samples, and the urban lettuce consumption data were collected through inquiry in Maputo Metropolis covering 350 persons. A non-parametric bootstrap was performed involving 10,000 iterations over the collected dataset, namely, water quality (E. coli) and lettuce consumption. The dose-response models were: Exponential for Cryptosporidium, Kummer Confluent hypergeomtric function (1F1) for Vibrio and Ascaris Gaussian hypergeometric function (2F1-(a,b;c;z) for norovirus. The annual infection risk estimates were performed using R 3.6.0 (CoreTeam) software by Monte Carlo (Latin hypercubes), a sampling technique involving 10,000 iterations. The annual infection risks values expressed by Median and the 95th percentile, per person per year (pppy) arising from the consumption of lettuce are as follows: Vibrio cholerae (1.00, 1.00), Cryptosporidium (3.91x10⁻³, 9.72x 10⁻³), nororvirus (5.22x10⁻¹, 9.99x10⁻¹) and Ascaris (2.59x10⁻¹, 9.65x10⁻¹). Thus, the consumption of the lettuce would result in greater risks than the tolerable levels ( < 10⁻³ pppy or 10⁻⁶ DALY) for all pathogens, and the Vibrio cholerae is the most virulent pathogens, according to the hit-single models followed by the Ascaris lumbricoides and norovirus. The sensitivity analysis carried out in this work pointed out that in the whole QMRA, the most important input variable was the reduction of pathogens (Spearman rank value was 0.69) between harvest and consumption followed by water quality (Spearman rank value was 0.69). The decision-makers (Mozambique Government) must strengthen the prevention measures related to pathogens reduction in lettuce (i.e., washing) and engage in wastewater treatment engineering.

Keywords: annual infections risk, lettuce, non-parametric bootstrapping, quantitative microbial risk assessment tools

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Authors: Rima Zakzouk, Noriko Hiraishi, Mohamed Mahdi Alshahni, Koichi Makimura, Junji Tagami

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Background and Purpose: Silver diamine fluoride (SDF) is used to prevent and arrest dental caries. The aim of this study is to evaluate the effect of SDF on reducing Candida albicans adhesion on dentin. Materials and Methods: Bovine dentin disks (6×6 mm) were cut by Isomet and polished using grit silicon carbide papers down to 2000 in order to obtain flat dentin surfaces. Samples were divided into two groups. The first group (SDF group) was treated with 38% SDF for 3 min, while the other group (control group) did not undergo SDF treatment. All samples were exposed to C. albicans suspension, washed after 6 hours incubation at 30 °C before to be tested using XTT (2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide) and real time PCR approaches. Statistical analyses of the results were performed at the significance level α = 0.05. Results: SDF inhibited C. albicans adhesion onto dentin. A significant difference was found between the SDF and control groups in both XTT and real time PCR tests. Conclusion: Using SDF to arrest the caries, could inhibit the Candida growth on dentin.

Keywords: silver diamine fluoride, dentin, real time PCR, XTT

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Authors: Daniel Thompson, Muhammad Peerbux, Sophie Cerutti, Hansraj Riteesh Bookun

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Patients with carotid stenosis, which may be asymptomatic or symptomatic in the form of transient ischaemic attack (TIA), amaurosis fugax, or stroke, often require an endarterectomy to reduce stroke risk. Risks of this procedure include stroke, death, myocardial infarction, and cranial nerve damage. Carotid endarterectomy is most commonly performed under general anaesthetic, however, it can also be undertaken with a regional anaesthetic approach. Our tertiary centre generally performs carotid endarterectomy under regional anaesthetic. Our major tertiary hospital mostly utilises regional anaesthesia for carotid endarterectomy. We completed a cross-sectional analysis of all cases of carotid endarterectomy performed under regional anaesthesia across a 10-year period between January 2010 to March 2020 at our institution. 350 patients were included in this descriptive analysis, and demographic details for patients, indications for surgery, procedural details, length of surgery, and complications were collected. Data was cross tabulated and presented in frequency tables to describe these categorical variables. 263 of the 350 patients in the analysis were male, with a mean age of 71 ± 9. 172 patients had a history of ischaemic heart disease, 104 had diabetes mellitus, 318 had hypertension, and 17 patients had chronic kidney disease greater than Stage 3. 13.1% (46 patients) were current smokers, and the majority (63%) were ex-smokers. Most commonly, carotid endarterectomy was performed conventionally with patch arterioplasty 96% of the time (337 patients). The most common indication was TIA and stroke in 64% of patients, 18.9% were classified as asymptomatic, and 13.7% had amaurosis fugax. There were few general complications, with 9 wound complications/infections, 7 postoperative haematomas requiring return to theatre, 3 myocardial infarctions, 3 arrhythmias, 1 exacerbation of congestive heart failure, 1 chest infection, and 1 urinary tract infection. Specific complications to carotid endarterectomy included 3 strokes, 1 postoperative TIA, and 1 cerebral bleed. There were no deaths in our cohort. This analysis of a large cohort of patients from a major tertiary centre who underwent carotid endarterectomy under regional anaesthesia indicates the safety of such an approach for these patients. Regional anaesthesia holds the promise of less general respiratory and cardiac events compared to general anaesthesia, and in this vulnerable patient group, calls for comparative research between local and general anaesthesia in carotid surgery.

Keywords: anaesthesia, carotid endarterectomy, stroke, carotid stenosis

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Authors: Seonggil Kim, Jeonghun Nam, Chae Seung Lim

Abstract:

Rapid diagnosis of fungal infections is critical for rapid antifungal therapy. However, it is difficult to detect extremely low concentration fungi in blood sample. To address the limitation, separation and concentration of fungi in blood sample are required to enhance the sensitivity of PCR analysis. In this study, we demonstrated a sheathless separation and concentration of fungi, candida cells using a viscoelastic fluid. To validate the performance of the device, microparticle mixture (2 and 13 μm) was used, and those particles were successfully separated based on the size difference at high flow rate of 100 μl/min. For the final application, successful separation of the Candida cells from the white blood cells (WBCs) was achieved. Based on the viscoelastic lateral migration toward the equilibrium position, Candida cells were separated and concentrated by center focusing, while WBCs were removed by patterning into two streams between the channel center and the sidewalls. By flow cytometric analysis, the separation efficiency and the purity were evaluated as ~99% and ~ 97%, respectively. From the results, the device can be the powerful tool for detecting extremely rare disease-related cells.

Keywords: candida cells, concentration, separation, viscoelastic fluid

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