BSL-3 Laboratory User Training Program at NUITM-KEMRI

Pathogens handled in a Biosafety Level 3 (BSL-3) containment laboratory pose significant risks to laboratory staff and the environment. It is therefore necessary to develop competency and proficiency among laboratory workers, and promote behaviors and practices that enhance safety through biosafety training. Following installation of our BSL-3 laboratory at the Center for Microbiology Research-Kenya Medical Research Institute, in 2006, a biosafety training program was developed to provide training on BSL-3 safety practices and procedures. The training program was developed based on the World Health Organization specifications, with adjustments to fit our research activities and biosafety needs. The program is composed of three phases namely; initial assessment, a training phase that includes theory and practicum, and final assessment. This article reports the content of our training program.

BSL-3 Laboratory User Training Program at NUITM-KEMRI

Pathogens handled in a Biosafety Level 3 (BSL-3) containment laboratory pose significant risks to laboratory staff and the environment. It is therefore necessary to develop competency and proficiency among laboratory workers and to promote appropriate behavior and practices that enhance safety through biosafety training. Following the installation of our BSL-3 laboratory at the Center for Microbiology Research-Kenya Medical Research Institute in 2006, a biosafety training program was developed to provide training on BSL-3 safety practices and procedures. The training program was developed based on World Health Organization specifications, with adjustments to fit our research activities and biosafety needs. The program is composed of three phases, namely initial assessment, a training phase including theory and a practicum, and a final assessment. This article reports the content of our training program.

The NUITM-KEMRI P3 Laboratory in Kenya: Establishment, Features, Operation and Maintenance

A biocontainment facility is a core component in any research setting due to the services it renders towards comprehensive biosafety observance. The NUITM-KEMRI P3 facility was set up in 2007 and has been actively in use since 2010 by researchers from this and other institutions. A number of hazardous agents have been handled in the laboratory among them MDR-TB and yellow fever viruses. The laboratory has the general physical and operational features of a P3 laboratory in addition to a number of unique features, among them the water-air filtration system, the eco-mode operation feature and automation of the pressure system that make the facility more efficient. It is equipped with biosafety and emergency response equipments alongside common laboratory equipments, maintained regularly using daily, monthly and yearly routines. Security and safety is strictly observed within the facility, enhanced by restricted entry, strict documentation and use of safety symbols. Training is also engrained within the operation of the laboratory and is undertaken and evaluated annually. Though the laboratory is in the process of obtaining accreditation, it is fully certified courtesy of the manufactures’ and constructed within specified standards.

[Study on molecular phylogeny of Schistosoma bovis based on mitochondrial DNA sequence and gene order].

OBJECTIVE: To determine the nucleotide sequence of the partial mitochondrial (mt) genome and the order of the mitochondrial protein-coding genes for Schistosoma bovis for analysis of possible phylogenetic position of this species in the genus Schistosoma. METHODS: The genomic DNA of adult worms were extracted by the GNT-K method. The target regions were amplified by PCR using a degenerated primer and specific primer. The PCR products were purified before ligating into the pGEM1 T-vector system. Recombinant plasmids were amplified in Escherichia coli, extracted and purified using routine methods. The nucleotide sequences were determined with an ABI PRISM 3100-Avant DNA sequencer using a BigDye Terminators v3.1 Cycle Sequencing Kit (Applied Bio-systems, CA, U.S.A.) with two T-vector specific primers (T7 and SP6). Positive colonies were sequenced with two internal specific primers to obtain the full sequence of each fragment on both strands by means of primer walking. Sequences of related schistosomes were retrieved from GenBank and aligned with our data. Gene trees were constructed using neighbor joining methods. RESULTS: The nucleotide sequence was determined and the gene order of this region in S. bovis was found as follows: NADHdehydrogenase4 (nad4)-trnQ (Gln)-trnK(Lys)-NADH dehydrogenase 3(nad3)-trnD (Asp)-NADH dehydrogenase 1(nad1). The gene order covering such region of S. bovis was similar to that of the African Schistosoma species, but strikingly different from the Asian species. Phylogenetic trees inferred from the alignment including partial nad4, nad3, partial nad1 and partial nad4+nad3+nad1 sequence for other 8 Schistosoma spp., respectively, revealed that S. bovis is placed proximally to S. haematobium in the African sub-group, which is identical with those placed by gene order in the African clade. CONCLUSION: The mtDNA analysis based on mitochondrial DNA sequence and the gene order strongly support the hypothesis that S. bovis belongs to the African schistosome clade rather than the Asian Schistosoma species.

THE EFFECT OF MEDICATION ON QUESTIONNAIRE ANALYSIS OF CHILDREN WITH SCHISTOSOMA MANSONI INFECTION IN TANZANIA

The effect of mass treatment on questionnaire results in the diagnosis of schistosomiasis mansoni was examined in 267 school children in an endemic area of Tanzania by Kato-Katz analysis of fecal specimens. The questionnaire asked for information about self-diagnosis, abdominal symptoms, blood in stools, history of wild water contact, stool examination and medication for schistosomiasis, and knowledge of the disease. A logistic regression analysis disclosed a significant association between schistosomiasis and "diarrhea" (p ≈ 0.007; odds ratio, 32.0; confidence interval, 2.5 - 403.3) and "abdominal enlargement" (p ≈ 0.003; odds ratio, 15.2; confidence interval, 2.6 - 90.1) among 61 children who had no history of medication for schistosomiasis. The sensitivity and specificity of the model were 86% and 64%, respectively. In contrast, no significant correlation was observed either for the 116 treated children, or for all the 267 children after the mass treatment. We conclude, therefore, that for children who had no history of medication for schistosomiasis, the questionnaire for abdominal manifestations provides reliable information on <I>S. mansoni</I> infection. However, once a child takes medication, the questionnaire becomes unreliable. This observation suggests that immunomodulation by anti-schistosomiasis drugs that kill adult worms exerts an effect on the appearance of abdominal manifestations and might explain the ambiguity of clinical symptoms in chronically infested patients, except in terminal cases. Further studies are required to develop a simple, rapid and cost-effective diagnostic method for monitoring <I>S. mansoni</I> infection after medication in local areas without resort to laboratory-based identification of schistosomiasis.

INCIDENCE OF TRAVELERS' DIARRHEA AMONG JAPANESE VISITING THAILAND

A cross-sectional survey of 327 Japanese short-term travelers (≤3 weeks) arriving in Bangkok, Thailand was conducted to assess the incidence of travelers’ diarrhea (TD) as well as their symptoms and treatment-seeking behaviors. The incidence of the first episode of TD (FTD) was ascertained retrospectively by questionnaire. Reported by 69 travelers, FTD clustered within the first 8 days of arrival in Thailand, and the incidence rate varied from 2% to 8% with the highest incidence on the third day.<BR> Cumulative probability of FTD was 19% for those arriving in Thai directly from Japan, 42 % for those arriving via Southeast Asia, and 25% for those arriving via other regions at Day 7 by the Kaplan-Meier survival analysis. Log rank test revealed a higher FTD risk for travelers arriving via other Southeast Asian countries than for those arriving directly from Japan (<I>P</I> < 0.005). Of all the 69 FTD episodes, 33% had classic TD defined as ≥3 unformed stools per 24 hours with at least one accompanying symptom, 49% had moderate TD defined as ≤2 unformed stools with at least one additional symptom or more unformed stools without additional symptoms, and 17% had mild TD defined as with ≤ 2 unformed stools without additional symptoms. Cumulative probability of FTD at Day 7 was 12% for classic TD, 25% for classic plus moderate TD and 30% for all the TD. More than 38% of travelers with diarrhea took medicine brought from Japan. Among travelers with classic TD, 35% bought medicine in Thailand, whereas 47-50% of travelers with moderate and mild TD took only rest without any treatment.