Rotavirus Disease Burden and Molecular Epidemiology in Children with Acute Diarrhoea Age Less than 5 Years in Nepal.

Introduction: Rotavirus is the most common cause of severe gastroenteritis in infants and young children worldwide. The aim of the study was to determine rotavirus disease burden and distribution of rotavirus genotypes in children less than five years of age. Methodology: Stool samples were collected from 1768 hospital admitted children under 5 years of age with acute watery diarrhea during November 2005 to October 2008. Rotavirus in stool samples was detected by Enzyme Immuno Assay (EIA) and positive specimens for rotavirus positive was genotyped by reverse transcription-polymerase chain reaction (RTPCR) and sequencing methods. Results: The prevalence of rotavirus was 36.59% (647/1768). Detection was higher in male (33.3%) than female (26.9%). The prevalence was higher during the month of January (2005-06) and February in other periods. During 2005-06, high prevalence of rotavirus was noted in children of age between 6-11 months while in 2006-07 and 2007-08 high prevalence was seen in age group 12-23 months. The most common genotyping of rotavirus identified 5 G types and 3 P types. Genotypes G12 and P [8] were most common during both periods studied (G12; 50%, 29% 33.7% in 2005-06, 2006-07 and 2007-08 respectively and P[8]; 47%, 35% and 43.5% in 2005-06, 2006-07 and 2007-08 respectively). Among six combined genotypes, G12P [6] was most prevalent (34%, 24% and 47.5% in 2005-06, 2006-07 and 2007-08 respectively) where as G1P [6] (4%) was seen in year 2007- 08 only. Conclusions: The study demonstrates the burden of rotavirus diarrhea in Nepal. The data on rotavirus genotypes will help inform decisions as to whether rotavirus vaccine should be considered for inclusion in to Nepal’s National Immunization Program.

Rotavirus Nosocomial Infection in Children under 5 years of age: A Preliminary study in Nepal.

Introduction: Rotavirus is one of the most common causes of acute gastroenteritis among infants and young children. The spread of rotavirus infection in pediatric wards can cause acute diarrhoea during hospitalization, and in turn, prolong hospitalization or need, rehospitalization. The aim of the study was to determine the prevalence of nosocomial infection due to rotavirus and other causal organisms among children less than 5 years of age. Methodology: A cross-sectional study between November 2009 and May 2010 was conducted among 96 pediatric patients in children hospital, Nepal. Stool samples were collected during hospitalization and up to 3 days after discharge from the hospital. Rotavirus antigens were detected by EIA and standard microbiological procedure were applied for other enteric pathogens. Results: The prevalence of nosocomial infection due to rotavirus was 30.2% (29/96). Of them, 16 (55.17%) of the 29 children with nosocomial infection during hospitalization and/or up to 72 hours after discharge developed diarrhoea. The prevalence of symptomatic cases of nosocomial infection was 55.17% (16/29), which involved 16.7% (16/96) of the study subjects, showing a prevalence of 44.82% (13/29) for asymptomatic nosocomial infection consisted of 13.54% (13/96) of the total subjects. Eleven of the 16 symptomatic subjects of nosocomial infection were affected by diarrhoea during hospitalization and five subjects after discharge. Bacteria (8.3%) and protozoa (5.2%) agents were also causing nosocomial infection in children. Conclusions: Children attending hospital may asymptomatically carry enteric pathogens and potentially act as source of nosocomial infection. Due to the relatively high frequency of nosocomial infection in the Children Hospital of Nepal, it is necessary to follow strictly hygienic rules such as isolation of patients with diarrhoea and hand-washing before and after the examination of each patient and cleaning the examination instruments after use in each patient.

A Perspective Study on the Etiology of Diarrhea in Children Less than 12 Years of age attending Kanti Children’s Hospital.

Introduction: Diarrhea is the most common illness among children causing highest number of mortality and morbidity in the developing countries. Objective: This study was conducted to determine the etiological agents of diarrhea in children less than 12 years of age. Methods: The study was carried out in Tribhuvan University Teaching Hospital, Health Research Laboratory and stool specimen were collected from Kanti Children’s Hospital between February 2007 and August 2007. The specimens were processed by standard microbiological methods, serological diagnosis for the complete identification of bacterial isolate and use of Rotaclone for diagnosis for Rotavirus. Results: A total 500 specimen were processed and 312 (62.4%) cases were identified with enteropathogens. Out of 500, 165 (33%) showed significant bacterial growth, 110 (22%) single or multiple parasitic infestation, and 167 (21.4%) Rotavirus. Among the bacterial isolate, Escherichia. coli (20.2%) were highest in number, Entamoeba. histolytica (10%), among the protozoa and Ascaris lumbricoide (1.4%) among the helminth. Age group 0-2 years showed most number of cases. The prevalence of Rotavirus infection was more in Inpatient (65.4%) than Outpatient (34.6%) (P>0.05). Ethnicgroup wise distribution showed that Gurung/Magar/ Rai/Tamang were highly infected (34.6%) (P>0.05). Conclusion: The results showed that bacteria were the major etiological agents of diarrhea in children than parasites and Rotavirus. E. coli among the bacteria, E. histolytica among the parasite and Rotavirus constituted the major causative agents identified. The age group 0-2years was the most vulnerable group where most of the enteropathogens were detected.

Gender variations in the prevalence of parasitic infections and the level of awareness in adolescents in rural Nepal.

The study was conducted in rural school adolescent children to investigate the awareness and its association in parasitic infections in boys and girls. Of the 182 children examined 119 (65.3%) were male and 63 (34.6%) were female, age ranged 12-20 years with median age 15 years. Out of 182 stool samples examined 73 (40%) were found to be positive for parasites in which two or more parasites were found in 10 stool samples. Giardia lamblia 33 (18.1%) was the predominant parasite followed by hook worm 27 (14.8%) Entamoeba histolytica 13 (7.1%), Ascaris lumbricoides 05 (2.7%), Hyminolepis nana 02 (2.2%) and Trichuris trichiuria 01 (0.5%). Thirty-one (49.2%) in 63 females and 40 (33.6%) of the 119 males were positive for parasitic infections. The prevalence of worm infection was significantly higher in female children than male (p < or = 0.05). In contrast to the high parasitic prevalence rate in females they possessed significantly higher levels of awareness about parasitic infections. Out of 119 males 99 (83.2%) and 61 (96.8%) of the 63 females (p < or = 0.05) knew that worms suck food from host body. Similarly, 62.2% of males and 96.85 of females (p < or = 0.05) knew that parasites suck blood from human body. More study should be carried out to find out the gender difference in parasitic infection and level of exposure to risk factors.

Emerging pathogen Cyclospora cayetanensis infection in Nepal.

Cylospora cayetanensis, an emerging parasitic pathogen of human is being increasingly recognized throughout the world, however the means of transmission and the possibility of a reservoir host remain an enigma. A longitudinal study on cyclosporiasis in different parts of Nepal was carried out from April, 1995 until November, 2000. Fecal specimens were collected from symptomatic and asymptomatic patients. The data shows a distinct seasonality with the highest infection rates occurring during the summer and rainy season of the year. Attempts have been made to determine the sources of infection and possible reservoir hosts. Stools were examined from nearly 700 animals such as chickens, pigs, buffalos, cows, dogs, cats, monkeys, rats, mice and pigeons. In addition, vegetable farms around the Kathmandu Valley were examined during the seasonal high and low periods of transmission. C. cayetanensis-like oocysts were found in sewage water and from vegetable washings on five occasions during June, July, August, October, and November. Similarly, C. cayetanensis-like oocysts were recovered from mice, rats, chickens, and dogs. These results suggest that these sources may be important in the transmission of this parasitosis. However, further studies will be required to obtain definitive answers on transmission.

Study of Cyclospora cayetanensis in health care facilities, sewage water and green leafy vegetables in Nepal.

Cyclospora cayetanensis, a newly emerging parasite, is endemic in Nepal. A total of 2,123 stool specimens were collected from 3 health care facilities based on clinical symptoms during the period between 1995 to October, 1998. Out of these specimens, cayetanensis oocysts were found in 632 (29.8%). To identify possible sources for Cyclospora infection, drinking water, sewage water, green-leafy vegetables including fecal samples of various animals were collected and examined. The vegetable leaves were washed in distilled water then the washings, sewage water and drinking water were centrifuged and the sediment were examined microscopically. As a result, oocyst of Cyclospora were identified in sewage water and vegetable washings on four different occasions in June, August, October and November. The positive results were also confirmed as C. cayetanensis by development of 2 sporocysts after 2 week incubation period in potassium dichromate. A survey of 196 domestic animals from the same areas demonstrated that two chickens were positive for Cyclospora-like organism and others were negative. Although further studies are needed to clarify the direct link between Cyclospora infection and these sources, the results suggest that sewage water, green leafy vegetables are possible sources of infection and chickens could be possible reservoir host of Cyclospora in Nepal.

Monoclonal antibody-based ELISA for the detection of circulating Entamoeba histolytica antigen in hepatic amebiasis in hamsters (Mesocricetus auratus).

Circulating amebic antigens were determined by using a sandwich ELISA with specific monoclonal antibody in the sera of 35 group I hamsters, 7 of which were sacrificed at intervals after hepatic inoculation with 500,000 axenically grown HM:1:IMSS strain of E. histolytica trophozoites, 7 group II infected hamsters in which metronidazole treatment was given and 18 group III uninfected controls. Amebic antigenemia was demonstrated in 5 of 7 (71.4%), 6 of 7 (85.7%), 7 of 7 (100%), 7 of 7 (100%) and 7 of 7 (100%) of group I hamsters on days 5, 10, 15, 20 and 30 of infections respectively, whereas 6 of 7 (85.7%) of group II hamsters were weakly positive, one was negative and all 18 group III hamsters were negative. The sensitivity of the assay was 100% after the animals were infected 15 days onwards. The level of antigenemia in hamsters of group I with abscess was significantly higher than those of the same group without abscess (p < 0.05). Absence or reduction of antigenemia after treatment could be interpreted to mean a positive test of cure and favorable therapeutic response. The MAb-PAb-based ELISA for the detection of circulating E. histolytica represents a simple and sensitive diagnostic test for invasive amebiasis in hamsters. Application of this test in amebic liver abscess patients should be of diagnostic value for indication of present infection or test of cure after successful treatment.

Intestinal parasites in children and adults with and without abdominal discomfort from the Kathmandu area of Nepal.

The present study on intestinal parasites from the Kathmandu area of Nepal found that in subjectively healthy children (HC) and adults (HA) the total parasite load was 28.1% and 38.8%, respectively, whereas children (ADC) and adults (ADA) with abdominal discomfort had a load of 62.7% and 67.8%. The prevalence of nematodes in the 4 groups was significantly higher in those with abdominal discomfort, particularly of Hookworm, Enterobius, and Ascaris. Hymenolepis nana was the most common tapeworm, and also with the highest incidence in patients with abdominal complaints. Taenia solium and T. saginata were only found in the two adult groups, but with low prevalence rates. The highest incidence of Cryptosporidium was found in both groups with abdominal discomfort, notably among children. No "healthy" carriers were found among the adults, whereas the prevalence of "healthy" carriers among children corresponded to that of sick adults. The presence of Giardia was highest among the sick children, many "healthy" carriers among both children and adults were noted. E. histolytica and E. dispar had a surprisingly low prevalence in all 4 groups. In "healthy" children the parasite was twice as common than in sick, supporting earlier findings of morphologically identical species with and without pathogenicity. "Healthy" adults had the lowest prevalence and sick adults the highest in the 4 groups studies. Blastocystis hominis was most common among adults with abdominal complaints (2.8%). Trichomonas was also more common in this group, in which of 34 positive specimen, 28 were from women. In "healthy" children the prevalence of mixed parasite infection was 2.1%, in "healthy" adults more than 7%, while in sick children and adults the prevalence was 13.3% and 11.5%, respectively.