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Anthrax is a notorious disease of public health importance caused by Bacillus anthracis . The causative agent can also be used as a biological weapon. Spores of these bacteria can sustain extreme environmental conditions and remain viable in soil for decades. Domestic and wild ruminants are highly susceptible to this pathogen, which usually presents as a peracute to acute disease. In humans, cutaneous anthrax is frequent but pulmonary and enteric anthrax are more serious. Humans, animals, and the environment are all involved, making anthrax a perfect target for a One Health approach. The environment plays a key role in disease transmission. At a time when the One Health concept is not mere slogans, collaborative efforts of medical professionals, veterinarians, and environmental scientists will be valuable for the prevention and control of this disease. In this review, we discussed the transmission dynamics of anthrax in the environment, animals, and humans, as well as One Health strategies to control and prevent anthrax.

The abuse and misuse of antibiotics is one of the main drivers of antimicrobial resistance (AMR). Globally, AMR in food-producing animals is a significant public health concern. This study, therefore, assessed the knowledge, attitudes, and practices related to antibiotic usage (AMU) and AMR among poultry farmers in Nepal. We conducted a cross-sectional survey of 605 poultry farmers from six districts of Nepal from May to June 2022 to assess the status of knowledge, attitude, as well as practices toward prudent antibiotic usage (AMU) and AMR. The majority of the participants in our study were from the Chitwan district (31.6%; n = 191/605), aged 30-44 (54.2%; n = 328/605), males (70.4%; n = 426/605), and farmers with a higher secondary (28.76%; n = 174/605) level of education. The tetracyclines (28%, n = 228/828), aminoglycosides (23%, n = 188/828), and fluoroquinolones (15%, n = 126/828) were the most used antibiotics classes among poultry farmers. Although 87.8% ( n = 531/605) of poultry farmers used antibiotics, 49.8% ( n = 301/605) of them were aware of AMR, and 55.7% ( n = 337/605) knew that the misuse of antimicrobials could affect human and environmental health. There were significant differences in the knowledge, attitude, and practices toward prudent AMU and AMR among farmers who reared different birds. The mean knowledge, attitude, and practice score of the respondents were 7.81 ± 3.26, 5.8 ± 2.32, and 7.59 ± 3.38 when measured on a scale of 12, 10, and 15, respectively. Based on a cut-off of 75% of the maximum score, 49.4% ( n = 299/605), 62.8% ( n = 380/605), and 12.73% ( n = 77/605) of the respondents had good knowledge, attitude, and practices toward prudent AMU and AMR, respectively. The multivariable logistic regression analyses revealed that the positive predictors of good knowledge and attitude were male gender, higher level of education, district, and the types of birds (layers). Similarly, those of the male gender (OR: 3.36; 95% CI: 1.38-8.20; p = 0.008) and those that rear layers (OR: 4.63; 95% CI: 1.75-12.25; p = 0.003) were more likely to practice prudent usage of antimicrobials. The findings of this study show poor practice toward prudent antibiotic usage despite good knowledge of AMR. This study provides essential baseline data on the knowledge, attitudes, and practices of poultry farmers in Nepal and offers valuable insights that could help in the design of interventions and policies aimed at addressing illicit AMU and AMR in poultry in Nepal.

Avian pathogenic Escherichia coli (APEC) is an important extra-intestinal pathogenic E. coli (ExPEC), which often causes systemic infection in poultry and causes great economic loss to the breeding industry. In addition, as a major source of human ExPEC infection, the potential zoonotic risk of APEC has been an ongoing concern. Previous studies have pointed out that APEC is a potential zoonotic pathogen, which has high homology with human pathogenic E. coli such as uro-pathogenic E. coli (UPEC) and neonatal meningitis E. coli (NMEC), shares multiple virulence factors and can cause mammalian diseases. Previous studies have reported that O18 and O78 could cause different degrees of meningitis in neonatal rats, and different serotypes had different degrees of zoonotic risk. Here, we compared APEC DE205B (O2:K1) with NMEC RS218 (O18:K1:H7) by phylogenetic analysis and virulence gene identification to analyze the potential risk of DE205B in zoonotic diseases. We found that DE205B possessed a variety of virulence factors associated with meningitis and, through phylogenetic analysis, had high homology with RS218. DE205B could colonize the cerebrospinal fluid (CSF) of rats, and cause meningitis and nerve damage. Symptoms and pathological changes in the brain were similar to RS218. In addition, we found that DE205B had a complete T6SS, of which Hcp protein was its important structural protein. Hcp1 induced cytoskeleton rearrangement in human brain microvascular endothelial cells (HBMECs), and Hcp2 was mainly involved in the invasion of DE205B in vitro. In the meningitis model of rats, deletion of hcp2 gene reduced survival in the blood and the brain invasiveness of DE205B. Compared with WT group, Δhcp2 group induced lower inflammation and neutrophils infiltration in brain tissue, alleviating the process of meningitis. Together, these results suggested that APEC DE205B had close genetic similarities to NMEC RS218, and a similar mechanism in causing meningitis and being a risk for zoonosis. This APEC serotype provided a basis for zoonotic research.

Vaccination is the most effective preventive measure of COVID-19 available at present, but its success depends on the global accessibility of vaccines and the willingness of people to be vaccinated. As the vaccination rollouts are increasing worldwide, it is important to assess public perception and willingness towards vaccination, so that the aim of mass vaccination will be successful. This study aimed to understand public perception towards COVID-19 vaccines and their willingness to get vaccinated in Nepal. This cross-sectional online survey was conducted among 1196 residents of Nepal in August 2021; most of the participants of this online survey were young adults (18-47 years) with university-level education. A total of 64.5% (771/1196) of the participants perceived COVID-19 vaccines to be safe and risk-free, while 68.6% (820/1196) agreed that vaccination would be efficient in the fight against this pandemic. Most of the participants (841/1196, 70.3%) disagreed that people are getting COVID-19 vaccines easily in Nepal, while they agree with the prioritization of older adults and healthcare workers for vaccination. A total of 61.1% (731/1196) of the participants had received at least one dose of the vaccine. Among the unvaccinated, 93.3% (434/465) were willing to get vaccinated when their turn came. The higher confidence of younger adults in vaccines and the vaccination process is encouraging, as that can help educate others who are hesitant or are not positive towards the idea of receiving vaccines. Dissemination of correct and current information, acquisition of enough doses of vaccines, and equitable distribution of vaccines will be required to achieve successful completion of the COVID-19 vaccination campaign in Nepal.

Acanthamoeba Keratitis (AK) can lead to substantial vision loss and morbidity among contact lens wearers. Misdiagnosis or delayed diagnosis is a major factor contributing to poor outcomes of AK. This study aimed to assess the effect of two antibiotics and one anaesthetic drug used in the diagnosis and nonspecific management of keratitis on the autofluorescence patterns of Acanthamoeba and two common bacteria that may also cause keratitis. Acanthamoeba castellanii ATCC 30868, Pseudomonas aeruginosa ATCC 9027, and Staphylococcus aureus ATCC 6538 were grown then diluted in either PBS (bacteria) or ¼ strength Ringer's solution ( Acanthamoeba ) to give final concentrations of 0.1 OD at 660 nm or 10 4 cells/mL. Cells were then treated with ciprofloxacin, tetracycline, tetracaine, or no treatment (naïve). Excitation-emission matrices (EEMs) were collected for each sample with excitation at 270-500 nm with increments in 5 nm steps and emission at 280-700 nm at 2 nm steps using a Fluoromax-4 spectrometer. The data were analysed using MATLAB software to produce smoothed color-coded images of the samples tested. Acanthamoeba exhibited a distinctive fluorescence pattern compared to bacteria. The addition of antibiotics and anaesthetic had variable effects on autofluorescence. Tetracaine altered the fluorescence of all three microorganisms, whereas tetracycline did not show any effect on the fluorescence. Ciprofloxacin produced changes to the fluorescence pattern for the bacteria, but not Acanthamoeba . Fluorescence spectroscopy was able to differentiate Acanthamoeba from P. aeruginosa and S. aureus in vitro. There is a need for further assessment of the fluorescence pattern for different strains of Acanthamoeba and bacteria. Additionally, analysis of the effects of anti-amoebic drugs on the fluorescence pattern of Acanthamoeba and bacteria would be prudent before in vivo testing of the fluorescence diagnostic approach in the animal models.

Acanthamoeba , an opportunistic pathogen is known to cause an infection of the cornea, central nervous system, and skin. Acanthamoeba feeds different microorganisms, including potentially pathogenic prokaryotes; some of microbes have developed ways of surviving intracellularly and this may mean that Acanthamoeba acts as incubator of important pathogens. A systematic review of the literature was performed in order to capture a comprehensive picture of the variety of microbial species identified within Acanthamoeba following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. Forty-three studies met the inclusion criteria, 26 studies (60.5%) examined environmental samples, eight (18.6%) studies examined clinical specimens, and another nine (20.9%) studies analysed both types of samples. Polymerase chain reaction (PCR) followed by gene sequencing was the most common technique used to identify the intracellular microorganisms. Important pathogenic bacteria, such as E. coli , Mycobacterium spp. and P. aeruginosa, were observed in clinical isolates of Acanthamoeba , whereas Legionella , adenovirus, mimivirus, and unidentified bacteria ( Candidatus ) were often identified in environmental Acanthamoeba . Increasing resistance of Acanthamoeba associated intracellular pathogens to antimicrobials is an increased risk to public health. Molecular-based future studies are needed in order to assess the microbiome residing in Acanthamoeba , as a research on the hypotheses that intracellular microbes can affect the pathogenicity of Acanthamoeba infections.

Background: The prevalence of disinfectant resistance in Pseudomonas aeruginosa is on the rise. P. aeruginosa is the most common bacteria isolated from cases of microbial keratitis. Many multi-purpose contact lens disinfectant solutions are available to decontaminate contact lenses before use and to help reduce the incidence of infections. However, with increasing disinfectant resistance, the effect of multi-purpose disinfectant solutions may diminish. The goal of this study was to examine genes associated with disinfectant resistance in ocular isolates of P. aeruginosa and understand the strain's susceptibility to different multipurpose disinfectant solutions., Methods: Seven potential disinfectant resistance genes were used in BLASTn searches against the whole genomes of 13 eye isolates of P. aeruginosa . A microdilution broth method was used to examine susceptibility to four different multipurpose disinfectant solutions., Results: All strains possessed the sugE2 , sugE3 and emrE ( qacE ) genes. The sugE1 and qacEdelta1 genes were present in 6/13 isolates. No strains contained the qacF or qacG genes. All tested disinfectant solutions had the ability to kill all test strains at 100% concentration, with some strains being susceptible at 1:8 dilutions of the disinfecting solutions. However, the presence of disinfectant resistance genes was not associated with susceptibility to multi-purpose disinfectants., Conclusion: All four tested contact lens disinfectant preparations are effective against P. aeruginosa isolates regardless of the presence of disinfectant resistance genes.