Dynamics of MDR Enterobacter cloacae outbreaks in a neonatal unit in Nepal: insights using wider sampling frames and next-generation sequencing.

OBJECTIVES: There are limited data on Enterobacter cloacae outbreaks and fewer describing these in association with NDM-1. With whole-genome sequencing, we tested the hypothesis that a cluster of 16 E. cloacae bacteraemia cases in a Nepali neonatal unit represented a single clonal outbreak, using a wider set of epidemiologically unrelated clinical E. cloacae isolates for comparison. METHODS: Forty-three isolates were analysed, including 23 E. cloacae and 3 Citrobacter sp. isolates obtained from blood cultures from 16 neonates over a 3 month period. These were compared with two contemporaneous community-associated drug-resistant isolates from adults, a unit soap dispenser isolate and a set of historical invasive isolates (n=14) from the same geographical locality. RESULTS: There were two clear neonatal outbreaks and one isolated case in the unit. One outbreak was associated with an NDM-1 plasmid also identified in a historical community-associated strain. The smaller, second outbreak was likely associated with a contaminated soap dispenser. The two community-acquired adult cases and three sets of historical hospital-associated neonatal isolates represented four additional genetic clusters. CONCLUSIONS: E. cloacae infections in this context represent several different transmission networks, operating at the community/hospital and host strain/plasmid levels. Wide sampling frames and high-resolution typing methods are needed to describe the complex molecular epidemiology of E. cloacae outbreaks, which is not appropriately reflected by routine susceptibility phenotypes. Soap dispensers may represent a reservoir for E. cloacae and bacterial strains and plasmids may persist in hospitals and in the community for long periods, sporadically being involved in outbreaks of disease.

Genome sequencing of an extended series of NDM-producing Klebsiella pneumoniae isolates from neonatal infections in a Nepali hospital characterizes the extent of community- versus hospital-associated transmission in an endemic setting.

NDM-producing Klebsiella pneumoniae strains represent major clinical and infection control challenges, particularly in resource-limited settings with high rates of antimicrobial resistance. Determining whether transmission occurs at a gene, plasmid, or bacterial strain level and within hospital and/or the community has implications for monitoring and controlling spread. Whole-genome sequencing (WGS) is the highest-resolution typing method available for transmission epidemiology. We sequenced carbapenem-resistant K. pneumoniae isolates from 26 individuals involved in several infection case clusters in a Nepali neonatal unit and 68 other clinical Gram-negative isolates from a similar time frame, using Illumina and PacBio technologies. Within-outbreak chromosomal and closed-plasmid structures were generated and used as data set-specific references. Three temporally separated case clusters were caused by a single NDM K. pneumoniae strain with a conserved set of four plasmids, one being a 304,526-bp plasmid carrying bla(NDM-1). The plasmids contained a large number of antimicrobial/heavy metal resistance and plasmid maintenance genes, which may have explained their persistence. No obvious environmental/human reservoir was found. There was no evidence of transmission of outbreak plasmids to other Gram-negative clinical isolates, although bla(NDM) variants were present in other isolates in different genetic contexts. WGS can effectively define complex antimicrobial resistance epidemiology. Wider sampling frames are required to contextualize outbreaks. Infection control may be effective in terminating outbreaks caused by particular strains, even in areas with widespread resistance, although this study could not demonstrate evidence supporting specific interventions. Larger, detailed studies are needed to characterize resistance genes, vectors, and host strains involved in disease, to enable effective intervention.

Investigation of the effects of hydrogenotrophic denitrification and anammox on the improvement of the quality of the drinking water supply system.

A drinking water supply system operates at Chyasal (in the Kathmandu Valley, Nepal) for purifying the groundwater that has high levels of ammonium nitrogen (NH4-N). However, high NO3-N concentrations were seen in the water after treatment. To further improve the quality of the drinking water, two types of attached growth reactors were developed for the purification system: (i) a hydrogenotrophic denitrification (HD reactor) and (ii) a concurrent reactor with anammox and hydrogenotrophic denitrification (AnHD reactor). For the HD reactor fed by water containing NO3-N, the denitrification efficiency was high (95-98%) for all NO3-N feed rates (20-40 mg/L). The nitrite-nitrogen (NO2-N) and nitrate-nitrogen (NO3-N) concentrations in the effluent were ∼0.5 mg/L. On the other hand, the AnHD reactor fed with water containing NH4-N and NO2-N was operated under varying flow rates of H2(30-70 mL/min) and intermittent supply periods (1-2 h). The efficiency of the anammox process was found to increase with decreasing H2flow rates or with increasing intermittency of the H2supply, while the efficiency of denitrification decreased under these conditions. For the optimal condition of 1.5 h intermittent H2supply, the anammox and denitrification efficiencies of the AnHD reactor reached 80% and 42%, respectively, while the concentrations of both NH4-N and NO2-N in the effluent were <1.0 mg/L, and no NO3-N was detected. From the experimental results, it is clear that both HD and AnHD reactors can function as efficient and critical units of the water purification system.

Bovine cerebral theileriosis: first molecular report in cross bred cattle calf in India.

Bovine tropical theileriosis caused by Theileria annulata, is a serious constraint to Indian dairy industry with more fatal infections in exotic cattle and substantial losses to cross-bred and indigenous zebu cattle. The present communication is to place on record the first report of molecular based confirmed case of cerebral theileriosis caused by T. annulata coupled with its morphological detection, clinical manifestations, haematological alterations and therapeutic management in a cross bred cattle calf from India. After preparation of peripheral thin blood smear from cross bred cattle calf at the site of collection and fixation with methanol, blood sample brought to Department of Veterinary Parasitology, College of Veterinary Science and A.H, Jabalpur and stained by standard protocol for Giemsa staining. Genomic DNA was isolated from the collected blood sample using QIAamp® DNA blood mini kit following the manufacturer's recommendations and PCR was performed. The cross bred cow calf revealed high rise in temperature (105.5°F), increased heart rate, labored breathing with seromucous nasal discharge, enlargement of prescapular lymph node and animal exhibited tonic clonic convulsions in response to any sudden noise. Giemsa stained thin blood smear revealed intraerythrocytic piroplasm and Koch'sblue bodies of T. annulata within the cytoplasm of lymphocytes. The species of Theileria was confirmed by molecular amplification of genomic DNA as T. annulata.

Clinical Evaluation of Nd:YAG Laser With and Without Dentin Bonding Agent for the Treatment of Occlusal Hypersensitivity.

PURPOSE: The aim of the present study was to evaluate and compare both the immediate and delayed desensitizing effects of the Nd:YAG (neodymium-doped:yttrium aluminum garnet; Nd3+:Y3Al5O12) laser with and without dentin bonding agent (DBA) on occlusal dentinal hypersensitivity (DH). METHODS AND MATERIALS: Twenty-one patients with a total of 117 chronic occlusal hypersensitive teeth were selected. Each subject had at least three hypersensitive teeth. These teeth were randomly allocated into three groups: group 1, DBA on the occlusal surface; group 2, Nd:YAG laser (1064-nm wavelength, 30 mJ of energy, 10 pulse/s, 60 seconds, two times); and group 3, Nd:YAG laser (1064-nm wavelength, 30 mJ of energy, 10 pulse/s, 60 seconds, two times) with DBA. Pain was assessed using a visual analog scale after stimulation of the sensitive teeth by using the sharp tip of an explorer and an air blast prior to treatment and immediately, one week, one month, and three months after treatment by one blinded examiner. RESULTS: A significant reduction in occlusal DH occurred at all time points in all of the experimental groups. The three groups showed significant improvements in discomfort immediately after treatment and after one week (p<0.001), but the Nd:YAG laser with DBA group had greater efficacy when compared with the other groups. The Nd:YAG laser group and Nd:YAG laser with DBA group had no significant differences at one month and three months after treatment (p>0.05); however, their desensitizing efficacy was superior to the DBA group. CONCLUSIONS: The Nd:YAG laser with DBA may be most effective in the long-term treatment of occlusal DH, although other measures also reduce DH.

Pubic lice infestation in man from Mhow, Madhya Pradesh.

Worldwide, 2-10% of human population is infested with pubic lice which are mostly found in the hair of pubic area, occasionally under the armpits, in the beard or mustache and on the eyebrows and eyelashes. They are usually transmitted through sexual contact or through toilet seats and bedding material. A twenty-year-old boy suffering from severe itching in the genital region during night hours submitted the lice for their identification in Department of Veterinary Parasitology, College of Veterinary Science and Animal Husbandry, Mhow. These lice were identified as Phthirus pubis.

Inter-field variability in the microbial communities of hydrothermal vent deposits from a back-arc basin.

Diverse microbial communities thrive on and in deep-sea hydrothermal vent mineral deposits. However, our understanding of the inter-field variability in these communities is poor, as limited sampling and sequencing efforts have hampered most previous studies. To explore the inter-field variability in these communities, we used barcoded pyrosequencing of the variable region 4 (V4) of the 16S rRNA gene to characterize the archaeal and bacterial communities of over 30 hydrothermal deposit samples from six vent fields located along the Eastern Lau Spreading Center. Overall, the bacterial and archaeal communities of the Eastern Lau Spreading Center are similar to other active vent deposits, with a high diversity of Epsilonproteobacteria and thermophilic Archaea. However, the archaeal and bacterial communities from the southernmost vent field, Mariner, were significantly different from the other vent fields. At Mariner, the epsilonproteobacterial genus Nautilia and the archaeal family Thermococcaceae were prevalent in most samples, while Lebetimonas and Thermofilaceae were more abundant at the other vent fields. These differences appear to be influenced in part by the unique geochemistry of the Mariner fluids resulting from active degassing of a subsurface magma chamber. These results show that microbial communities associated with hydrothermal vent deposits in back-arc basins are taxonomically similar to those from mid-ocean ridge systems, but differences in geologic processes between vent fields in a back-arc basin can influence microbial community structure.

Role of physicochemical properties in the estimation of skin permeability: in vitro data assessment by Partial Least-Squares Regression.

Skin provides passage for the delivery of drugs. The in vitro and in vivo testing of chemicals for estimation of dermal absorption is very time consuming, costly and has many ethical difficulties related to human and animal testing. The solution to the problem is Quantitative structure-permeability relationships. This method relates dermal penetration properties of a range of chemical compounds to their physicochemical parameters. In the present study, an effort has been made to develop models for the accurate prediction of skin permeability using a large, diverse dataset through the combination of various regression methods coupled with the Genetic Algorithm (GA)/Interval Partial Least-Squares Algorithm (iPLS). The descriptors were calculated using e-DRAGON and ADME Pharma Algorithms-Abrahams descriptors. The original dataset was divided into a training set and a testing set using the Kennard-Stone Algorithm. The selection of descriptors was made by the GA and iPLS. The model applicability domain was determined. The results showed that a three-parameter model built through Partial Least-squares Regression was most accurate with r(2) of 0.936.