Biological and genomic characterization of two newly isolated Elizabethkingia anophelis bacteriophages.

BACKGROUND: Elizabethkingia anophelis is an opportunistic pathogen that infects newborns and immunocompromised patients. Because the infection is associated with high mortality as a result of its intrinsic resistance to antibiotics, alternative treatment methods are needed. Our previous study successfully isolated the world's first E. anophelis phage, TCUEAP1, which showed beneficial protection to E. anophelis-infected mice. More new bacteriophages are needed in order to provide sufficient choices to combat E. anophelis infections. METHODS: In the current study, two new phages infecting E. anophelis were isolated from wastewater and were designated as TCUEAP2 and TCUEAP3. Further experiments, namely, transmission electron microscopy (TEM), infection assay, host-range analysis, and sequencing were performed to determine their biological and genomic characteristics. RESULTS: TEM analysis revealed that both TCUEAP2 and TCUEAP3 possess an icosahedral head with a non-contractile tail, and belong to the Siphoviridae family. Further experiments revealed that TCUEAP3 has a longer latent period and higher burst size compared to TCUEAP2. Host range analysis showed that both TCUEAP2 and TCUEAP3 have a narrow host range, infecting only their respective hosts. The genomic size of phage TCUEAP2 was 42,403 bps containing 61 predicted open reading frames (ORFs), whereas the genome size of TCUEAP3 was 37,073 bps containing 40 predicted ORFs. CONCLUSION: Due to the distinct biological characteristics of TCUEAP2 and TCUEAP3, they may be satisfactory for clinical uses such as preparation of phage cocktails or decontamination in clinical settings.

Isolation and Characterization of a New Phage Infecting Elizabethkingia anophelis and Evaluation of Its Therapeutic Efficacy in vitro and in vivo.

Elizabethkingia spp. are a group of non-fermentative, Gram-negative, catalase-positive, and non-motile bacilli. They can cause meningitis in neonates and immunosuppressed patients, and lead to high mortality. Considering the rising trend of drug resistance among bacteria pathogens, bacteriophage (phage) therapy is a potential alternative to antibiotics for treating multidrug-resistant bacterial infections. However, so far, no phages specific for Elizabethkingia spp. have been reported. Using a clinically isolated Elizabethkingia anophelis as the host, the phage TCUEAP1 was isolated from wastewater of Hualien Tzu Chi hospital. The phage particle of TCUEAP1 under electron microscopy was revealed to belong to the siphoviridae family, with a head size of 47 nm, and a tail dimension 12 nm in diameter and 172 nm in length. The one-step growth analysis showed that the latent period of TCUEAP1 was about 40 min with a rise period lasting about 20 min, yielding a burst size of approximately 10 PFU/cell. The adsorption rate of TCUEAP1 reached about 70% in 20 min. Using 20 isolates of Elizabethkingia spp. to test the host range of TCUEAP1, it displayed narrow spectrum infecting three strains of E. anophelis, but forming spot lysis on 16 strains. The sequence result showed that the genome of TCUEAP1 is a double-stranded DNA of 49,816 bp, containing 73 predicted open reading frames. Further genomic analysis showed TCUEAP1 to be a new phage with no resemblance to publicly available phage genomes. Finally, in a mouse intraperitoneal infection model, at 6 h after the bacterial injection, TCUEAP1 decreased the bacterial load by fivefold in blood. Also, TCUEAP1 rescued 80% of mice heavily infected with E. anophelis from lethal bacteremia. We hope that the isolation and characterization of TCUEAP1, the first phage infecting Elizabethkingia spp., can promote more studies of the phages targeting this newly emerging bacterial pathogen.

Uttarakhand Disaster 2013: A Report on Psychosocial Adversities Experienced by Children and Adolescents.

OBJECTIVE: To report the psychosocial adversities faced by children and adolescents in the Uttarkashi, district of Uttarakhand, experienced immediately after the Himalayan Tsunami in June 2013. Also to discuss issues pertinent to the disaster management including the needs of the disaster affected areas and future challenges. METHODS: This is a cross sectional observational report from the community assessment and interventions that were carried out as part of the disaster relief work by National Institute of Mental Health and Neurosciences (NIMHANS), Bangalore within 1 mo of the disaster. Assessments and interventions were done by a team consisting of psychiatry resident, clinical psychologist, psychiatric social worker and a nurse. All diagnosis were made using International Classification of Diseases 10 (ICD 10) and the data was analysed using descriptive statistics and chi-square tests. RESULTS: A total of 300 children were screened; the mean age of the sample was 11.5 y and 65(32.5%) were boys. Two hundred (66.7%) children/adolescents reported one or the other psychosocial adversities attributable to the disaster. Psychological distress was present in 54/300 (18%) of the individuals. Loss of shelter and loss of playing space were the social issues having a statistically significant association with psychological distress signals such as feelings of anxiety, helplessness, insecurity, grief and uncertainty. Stress induced diagnosable psychiatric disorder was not present in any child or adolescent, however stress related psychiatric symptoms were present in around 13%. CONCLUSIONS: Himalayan tsunami of Uttarakhand in 2013 was associated with considerable psychosocial adversities among the resident children and adolescents. As children are a vulnerable population, a public health approach towards assessment and management of the psychosocial adversities in this population is urgently required at the state and national levels.

Psychiatric and medical disorders in the after math of the uttarakhand disaster: assessment, approach, and future challenges.

PURPOSE: To present the descriptive data on the frequency of medical and psychiatric morbidity and also to discuss various pertinent issues relevant to the disaster management, the future challenges and psychosocial needs of the 2013 floods in Uttarakhand, India. MATERIALS AND METHODS: Observation was undertaken by the disaster management team of National Institute of Mental Health and Neurosciences in the worst affected four districts of Uttarakhand. Qualified psychiatrists diagnosed the patients using the International Classification of Diseases-10 criteria. Data were collected by direct observation, interview of the survivors, group sessions, individual key-informant interview, individual session, and group interventions. RESULTS: Patients with physical health problems formed the majority of treatment seekers (39.6%) in this report. Only about 2% had disaster induced psychiatric diagnoses. As was expected, minor mental disorders in the form of depressive disorders and anxiety disorders formed majority of the psychiatric morbidity. Substance use disorders appear to be very highly prevalent in the community; however, we were not able to assess the morbidity systematically. CONCLUSIONS: The mental health infrastructure and manpower is abysmally inadequate. There is an urgent need to implement the National Mental Health Program to increase the mental health infrastructure and services in the four major disaster-affected districts.