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A comparative study of four methods for detecting Giardia lamblia in children with diarrheal disease and malabsorption

Kamath KR, Murugasu R

Gastroenterology, 1974 Jan;66(1):16-21.
PMID: 4809494
Matched MeSH terms: Feces/microbiology
Fulltext A pentaplex PCR assay for the detection and differentiation of Shigella species

Ojha SC, Yean Yean C, Ismail A, Singh KK

Biomed Res Int, 2013;2013:412370.
PMID: 23509722 DOI: 10.1155/2013/412370

The magnitude of shigellosis in developing countries is largely unknown because an affordable detection method is not available. Current laboratory diagnosis of Shigella spp. is laborious and time consuming and has low sensitivity. Hence, in the present study, a molecular-based diagnostic assay which amplifies simultaneously four specific genes to identify invC for Shigella genus, rfc for S. flexneri, wbgZ for S. sonnei, and rfpB for S. dysenteriae, as well as one internal control (ompA) gene, was developed in a single reaction to detect and differentiate Shigella spp. Validation with 120 Shigella strains and 37 non-Shigella strains yielded 100% specificity. The sensitivity of the PCR was 100 pg of genomic DNA, 5.4 × 10(4) CFU/ml, or approximately 120 CFU per reaction mixture of bacteria. The sensitivity of the pentaplex PCR assay was further improved following preincubation of the stool samples in gram-negative broth. A preliminary study with 30 diarrhoeal specimens resulted in no cross-reaction with other non-Shigella strains tested. We conclude that the developed pentaplex PCR assay is robust and can provide information about the four target genes that are essential for the identification of the Shigella genus and the three Shigella species responsible for the majority of shigellosis cases.

Matched MeSH terms: Feces/microbiology
A shellfish-borne cholera outbreak in Malaysia

Dutt AK, Alwi S, Velauthan T

Trans R Soc Trop Med Hyg, 1971;65(6):815-8.
PMID: 5157442
Matched MeSH terms: Feces/microbiology
A signal-amplified electrochemical DNA biosensor incorporated with a colorimetric internal control for Vibrio cholerae detection using shelf-ready reagents

Low KF, Zain ZM, Yean CY

Biosens Bioelectron, 2017 Jan 15;87:256-263.
PMID: 27567251 DOI: 10.1016/j.bios.2016.08.064

A novel enzyme/nanoparticle-based DNA biosensing platform with dual colorimetric/electrochemical approach has been developed for the sequence-specific detection of the bacterium Vibrio cholerae, the causative agent of acute diarrheal disease in cholera. This assay platform exploits the use of shelf-stable and ready-to-use (shelf-ready) reagents to greatly simplify the bioanalysis procedures, allowing the assay platform to be more amenable to point-of-care applications. To assure maximum diagnosis reliability, an internal control (IC) capable of providing instant validation of results was incorporated into the assay. The microbial target, single-stranded DNA amplified with asymmetric PCR, was quantitatively detected via electrochemical stripping analysis of gold nanoparticle-loaded latex microspheres as a signal-amplified hybridization tag, while the incorporated IC was analyzed using a simplified horseradish peroxidase enzyme-based colorimetric scheme by simple visual observation of enzymatic color development. The platform showed excellent diagnostic sensitivity and specificity (100%) when challenged with 145 clinical isolate-spiked fecal specimens. The limits of detection were 0.5ng/ml of genomic DNA and 10 colony-forming units (CFU)/ml of bacterial cells with dynamic ranges of 0-100ng/ml (R(2)=0.992) and log10 (1-10(4) CFU/ml) (R(2)=0.9918), respectively. An accelerated stability test revealed that the assay reagents were stable at temperatures of 4-37°C, with an estimated ambient shelf life of 200 days. The versatility of the biosensing platform makes it easily adaptable for quantitative detection of other microbial pathogens.

Matched MeSH terms: Feces/microbiology*
A strain of urease-producing Vibrio parahaemolyticus isolated in Malaysia

Jegathesan M, Paramasivam T

J Diarrhoeal Dis Res, 1985 Sep;3(3):162.
PMID: 3833915
Matched MeSH terms: Feces/microbiology
A thermostabilized magnetogenosensing assay for DNA sequence-specific detection and quantification of Vibrio cholerae

Low KF, Karimah A, Yean CY

Biosens Bioelectron, 2013 Sep 15;47:38-44.
PMID: 23545172 DOI: 10.1016/j.bios.2013.03.004

Vibrio cholerae is a human pathogen that causes mild to severe diarrheal illnesses and has major public health significance. Herein, we present a thermostabilized electrochemical genosensing assay combining the use of magnetic beads as a biorecognition platform and gold nanoparticles as a hybridization tag for the detection and quantification of V. cholerae lolB gene single-stranded asymmetric PCR amplicons as an alternative to the time-consuming classical isolation method. This thermostabilized, pre-mixed, pre-aliquoted and ready-to-use magnetogenosensing assay simplified the procedures and permitted the reaction to be conducted at room temperature. The asymmetric PCR amplicons were hybridized to a magnetic bead-functionalized capture probe and a fluorescein-labeled detection probe followed by tagging with gold nanoparticles. Electrochemical detection of the chemically dissolved gold nanoparticles was performed using the differential pulse anodic stripping voltammetry method. The real-time stability evaluation of thermostabilized assay was found to be stable for at least 180 days at room temperature (25-30°C). The analytical specificity of the assay was 100%, while its analytical sensitivity was linearly related to different concentrations of 200-mer synthetic target, purified genomic DNA, and bacterial culture with a limit of detection (LoD) of 3.9nM, 5pg/µl, and 10(3)CFU/ml, respectively. The clinical applicability of the assay was successfully validated using spiked stool samples with an average current signal-to-cut-off ratio of 10.8. Overall, the precision of the assay via relative standard deviation was <10%, demonstrating its reliability and accuracy.

Matched MeSH terms: Feces/microbiology
Acclimatisation-induced stress influenced host metabolic and gut microbial composition change

Yap IK, Kho MT, Lim SH, Ismail NH, Yam WK, Chong CW

Mol Biosyst, 2015 Jan;11(1):297-306.
PMID: 25382376 DOI: 10.1039/c4mb00463a

Understanding the basal gut bacterial community structure and the host metabolic composition is pivotal for the interpretation of laboratory treatments designed to answer questions pertinent to host-microbe interactions. In this study, we report for the first time the underlying gut microbiota and systemic metabolic composition in BALB/c mice during the acclimatisation period. Our results showed that stress levels were reduced in the first three days of the study when the animals were subjected to repetitive handling daily but the stress levels were increased when handling was carried out at lower frequencies (weekly). We also observed a strong influence of stress on the host metabolism and commensal compositional variability. In addition, temporal biological compartmental variations in the responses were observed. Based on these results, we suggest that consistency in the frequency and duration of laboratory handling is crucial in murine models to minimise the impact of stress levels on the commensal and host metabolism dynamics. Furthermore, caution is advised in consideration of the temporal delay effect when integrating metagenomics and metabonomics data across different biological matrices (i.e. faeces and urine).

Matched MeSH terms: Feces/microbiology
Additional oligofructose/inulin does not increase faecal bifidobacteria in critically ill patients receiving enteral nutrition: a randomised controlled trial

Majid HA, Cole J, Emery PW, Whelan K

Clin Nutr, 2014 Dec;33(6):966-72.
PMID: 24290345 DOI: 10.1016/j.clnu.2013.11.008

Patients with diarrhoea during enteral nutrition (EN) have been shown to have low faecal bifidobacteria concentrations. Oligofructose/inulin selectively stimulate the growth of bifidobacteria in healthy humans. This study investigates the effect of additional oligofructose/inulin on the gastrointestinal microbiota, short-chain fatty acids (SCFA) and faecal output in patients receiving EN.

Matched MeSH terms: Feces/microbiology*
An appraisal of El Tor carrier state in Kelantan

Dutt AK, Tan Hock Joo

Med J Malaya, 1971 Mar;25(3):205-7.
PMID: 4253247
Matched MeSH terms: Feces/microbiology
An evaluation of the staphylococcal co-agglutination test for the detection of group A rotavirus in human faeces

Yap KL, Ooi YE, Khor CM, Wong SH

Malays J Pathol, 1992 Dec;14(2):105-10.
PMID: 1338997

The group A rotavirus staphylococcal co-agglutination test was evaluated and its sensitivity and specificity compared with an in-house enzyme-linked immunosorbent assay (ELISA) and a commercial latex agglutination test (Rotalex). In addition, the storage stability of the staphylococcal reagents was ascertained. Examination of 136 clarified suspensions of diarrhoeal faeces by the staphylococcal co-agglutination test revealed a high proportion of false positives (26%) and uninterpretable results (34%) due to non-specific agglutination. Non-specific agglutination could be removed effectively by prior absorption of the clarified faecal specimens with unsensitized staphylococci. The staphylococcal co-agglutination test was less sensitive and specific than the in-house enzyme-linked immunosorbent assay but was comparable to the Rotalex slide latex agglutination test. The staphylococcal reagents have a shelf life of at least 29 weeks.

Matched MeSH terms: Feces/microbiology*
An integrative study on biologically recovered polyhydroxyalkanoates (PHAs) and simultaneous assessment of gut microbiome in yellow mealworm

Ong SY, Kho HP, Riedel SL, Kim SW, Gan CY, Taylor TD, et al.

J Biotechnol, 2018 Jan 10;265:31-39.
PMID: 29101024 DOI: 10.1016/j.jbiotec.2017.10.017

Polyhydroxyalkanoates (PHAs) are produced in microbes as a source of carbon and energy storage. They are biodegradable and have properties similar to synthetic plastics, which make them an interesting alternative to petroleum-based plastics. In this study, a refined method of recovering PHA from Cupriavidus necator biomass was proposed by incorporating the use of the yellow mealworm (the larval phase of the mealworm beetle, Tenebrio molitor) as partial purification machinery, followed by washing of the fecal pellets with distilled water and sodium hydroxide. The PHA contents of the cells used in this study were 55wt% (produced from palm olein) and 60 wt% (produced from waste animal fats). The treatment of distilled water and NaOH further increased the purity of PHA to 94%. In parallel, analysis of the 16S rRNA metagenomic sequencing of the mealworm gut microbiome has revealed remarkable changes in the bacterial diversity, especially between the mealworms fed with cells produced from palm olein and waste animal fats. This biological recovery of PHA from cells is an attempt to move towards a green and sustainable process with the aim of reducing the use of harmful solvents and strong chemicals during polymer purification. The results obtained show that - purities of >90%, without a reduction in the molecular weight, can be obtained through this integrative biological recovery approach. In addition, this study has successfully shown that the cells, regardless of their origins, were readily consumed by the mealworms, and there is a correlation between the feed type and the mealworm gut microbiome.

Matched MeSH terms: Feces/microbiology
An investigative study on the zoonotic potential of Helicobacter pylori

Shaaban SI, Talat D, Khatab SA, Nossair MA, Ayoub MA, Ewida RM, et al.

BMC Vet Res, 2023 Jan 21;19(1):16.
PMID: 36670434 DOI: 10.1186/s12917-023-03572-w

BACKGROUND: Helicobacter pylori is one of the most common bacterial infections and is widespread globally. It causes a variety of gastrointestinal disorders, though a great proportion of infections are asymptomatic. A total of 143 fresh stool samples were collected from apparently healthy farm and pet animals (43 cattle, 50 buffaloes, 50 sheep, 50 dogs, and 50 cats), in addition to 768 human stool samples. The samples were examined using stool antigen and rapid antibody tests, and further confirmation of glmM "human antigen-positive samples and animal milk samples" was conducted by polymerase chain reaction (PCR).
RESULTS: The prevalence rates of H. pylori infection in animals were 22.2% and 16% in antibody and stool antigen tests, respectively. The detection rates were 28%, 24%, 12%, 10%, and 4.7% in cats, dogs, buffaloes, sheep, and cattle, respectively. On the other hand, the prevalence rate of H. pylori infection in human stool samples was 74.8%, and a statistically significant association was observed between prevalence and several factors, such as sex, age, and locality. PCR was performed to detect the glmM gene of H. pylori, and this gene was found in 21 of 27 human antigen-positive samples and 5 of 13 animal milk samples.
CONCLUSIONS: H. pylori was detected in both human and animal samples. Furthermore, glmM was found in milk and human samples. Our findings suggest that pet and farm animals could transmit H. pylori infection to humans.

Matched MeSH terms: Feces/microbiology
Anaemia and iron deficiency anaemia among aboriginal schoolchildren in rural Peninsular Malaysia: an update on a continuing problem

Al-Mekhlafi MH, Surin J, Atiya AS, Ariffin WA, Mahdy AK, Abdullah HC

Trans R Soc Trop Med Hyg, 2008 Oct;102(10):1046-52.
PMID: 18617209 DOI: 10.1016/j.trstmh.2008.05.012

A cross-sectional study to determine the current prevalence of anaemia and iron deficiency anaemia (IDA) and to investigate the possible risk factors for IDA was carried out on 241 aboriginal schoolchildren (120 boys, 121 girls) aged 7-12 years and living in remote areas in Pos Betau, Pahang, Malaysia. Haemoglobin (Hb) level was measured and serum iron status was assessed by serum ferritin (SF), serum iron (SI) and total iron binding capacity measurements. Socioeconomic data were collected using pre-tested questionnaires. All children were screened for intestinal parasitic infections. Overall, 48.5% (95% CI 42.3-54.8) of children were anaemic (Hb<12 g/dl). The prevalence of IDA was 34% (95% CI 28.3-40.2), which accounted for 70.1% of the anaemia cases. The prevalence of IDA was significantly higher in females than males. Low levels of mothers' education and low household income were identified as risk factors for IDA. Severe trichuriasis also found to be associated with low SF and SI. Logistic regression confirmed low levels of mothers' education and gender as significant risk factors for IDA. Improvement of socioeconomic status and health education together with periodic mass deworming should be included in public health strategies for the control and prevention of anaemia and IDA in this population.

Matched MeSH terms: Feces/microbiology
Fulltext Annual and geographical distribution of Shigella serotypes in Sabah, Malaysia from 1974 to 1978

Kan SKP, Chan MKC

Med J Malaysia, 1980 Sep;35(1):9-13.
PMID: 7254007

19,983 cases of diarrhoea throughout Sabah, Malaysia from January 1971 to December 1978 were bacteriologically examined for Shigella. A total of 241 Shigella isolates representing 9 serotypes were encountered. S. flexneri and S. sonnei accounted for 69.7% and 29.5% of the isolates respectively. S. flexneri type 2 was very common and comprised 47% of the flexneri strains. S. flexneri types 5.6 and Y were rarely found. Only two cases of S. boydii were isolated. S. dysenteriae was not encountered. Isolation rates ranged from 0.64% to 1.73% while the percentages or cases of diarrhoea bacteriologically examined in relation to the number notified range from 13.7 to 29.6. Kota Kinabalu. Tawau and Sandakan accounted for 50.6%. 21.2% and 8.2% of Shigella isolates respectively. However. no isolations were made from Lahad Datu, Semporna and Victoria [Labuan Island]. S. flexneri type 5 was only found in Sandakan while S. flexnert type Y was isolated from Kota Kinubulu. No S. Sonnel was found in Ranau and Tenom.

Matched MeSH terms: Feces/microbiology
Antimicrobial and bactericidal impacts of Bacillus amyloliquefaciens CECT 5940 on fecal shedding of pathogenic bacteria in dairy calves and adult dogs

Vazquez-Mendoza P, Elghandour MMM, Alaba PA, Sánchez-Aparicio P, Alonso-Fresán MU, Barbabosa-Pliego A, et al.

Microb Pathog, 2018 Jan;114:458-463.
PMID: 29180295 DOI: 10.1016/j.micpath.2017.11.040

Two experiments were carried out to evaluate the bactericidal impacts of Bacillus amyloliquefaciens CECT 5940 on the shedding of faecal pathogenic bacteria in dairy calves (Experiment 1) and in adults dogs (experiment 2). In the calves experiment, a completely randomized design was used to investigate the faecal bacteria profile of Holstein dairy calves fed with either pasteurized waste milk (PWM; n = 9) or a formulated non-medicated milk replacer (NMR; n = 9) for 60 d. The NMR containing sodium-butyrate and the active probiotic B. amyloliquefaciens CECT 5940. In the dogs experiment, addition of same probiotic (i.e., B. amyloliquefaciens CECT 5940) was carried out in two stages. The first stage started from day 7-37, and the second from day 44-71. The assessment of faecal score measured on day 22, 37, 42, 57, 71 and 77 to determine the texture of the stools. Calves received PWM consumed (P

Matched MeSH terms: Feces/microbiology*
Antimicrobial susceptibility and pulsed: field Gel Electrophoretic analysis of Salmonella in a tertiary hospital in northern Malaysia

Thong KL, Lai WL, Dhanoa A

J Infect Public Health, 2011 Jun;4(2):65-72.
PMID: 21663875 DOI: 10.1016/j.jiph.2011.03.003

Salmonella infections remain a major public health problem in developing countries. The occurrence of infections caused by antimicrobial-resistant Salmonella has been on the rise complicating the available therapeutic options. The study aimed to determine the antibiograms and genotypes of prevalent Salmonella serotypes.

Matched MeSH terms: Feces/microbiology
Application of ribosomal RNA gene restriction patterns analysis and pulsed-field gel electrophoresis in distinguishing Salmonella weltevreden isolates in Malaysia

Goh YL, Puthucheary SD, Thong KL

Southeast Asian J. Trop. Med. Public Health, 2000 Dec;31(4):697-701.
PMID: 11414415

A representative sample of 20 isolates of Salmonella weltevreden strains from stool cultures of patients admitted at the University Hospital, Kuala Lumpur, Malaysia were analyzed. All the strains were susceptible to ampicillin, ceftriaxone, ciprofloxacin, chloramphenicol, tetracycline, trimethoprim, gentamicin and co-trimoxazole. Ribosomal RNA gene restriction pattern analysis of PstI-digested DNA gave three ribotypes while pulsed-field gel electrophoresis (PFGE) analysis of XbaI-digested DNA gave ten distinct profiles. PFGE was more discriminative than ribotyping in distinguishing the strains. The majority of the strains analyzed were very closely related with similarity coefficient values ranging from 0.8 to 1.0. Both PFGE and ribotyping could distinguish one of the strains which was obtained from a patient following a bone marrow transplant for beta-thalassemia major, indicating that this particular strain was unrelated to the rest of the strains from patients with acute gastroenteritis.

Matched MeSH terms: Feces/microbiology
Bifidobacterium longum BB536 alleviated upper respiratory illnesses and modulated gut microbiota profiles in Malaysian pre-school children

Lau AS, Yanagisawa N, Hor YY, Lew LC, Ong JS, Chuah LO, et al.

Benef Microbes, 2018 Jan 29;9(1):61-70.
PMID: 29065707 DOI: 10.3920/BM2017.0063

This 10-months randomised, double-blind, parallel and placebo-controlled study evaluated the effects of Bifidobacterium longum BB536 on diarrhoea and/or upper respiratory illnesses in 520 healthy Malaysian pre-school children aged 2-6 years old. The subjects randomly received a one-gram sachet containing either BB536 (5×109 cfu) or placebo daily. Data analysis was performed on 219 subjects who fully complied over 10-months (placebo n=110, BB536 n=109). While BB536 did not exert significant effects against diarrhoea in children, Poisson regression with generalised estimating equations model indicated significant intergroup difference in the mean number of times of respiratory illnesses over 10 months. The duration of sore throat was reduced by 46% (P=0.018), with marginal reduction for duration of fever (reduced by 27%, P=0.084), runny nose (reduced by 15%, P=0.087) and cough (reduced by 16%, P=0.087) as compared to the placebo. Principal coordinate analysis at genus level of the gut microbiota revealed significant differences between 0 and 10 months in the BB536 group (P<0.01) but not in placebo group (P>0.05). The abundance of the genus Faecalibacterium which is associated with anti-inflammatory and immuno-modulatory properties was significantly higher in the BB536 group (P<0.05) compared to the placebo group. Altogether, our present study illustrated the potential protective effects of BB536 against upper respiratory illnesses in pre-school Malaysian children, with gut microbiota modulating properties.

Matched MeSH terms: Feces/microbiology
Biological control of nematode parasites in sheep

Larsen M

J Anim Sci, 2006 Apr;84 Suppl:E133-9.
PMID: 16582084

In a world in which sheep producers are facing increasing problems due to the rapid spread of anthelmintic resistance, the battle against gastrointestinal parasitic nematodes is a difficult one. One of the potential new tools for integrated control strategies is biological control by means of the nematode-destroying microfungus Duddingtonia flagrans. This fungus forms sticky traps that catch developing larval stages of parasitic nematodes in the fecal environment. When resting spores (chlamydospores) of this fungus are fed daily to grazing animals for a period of time, the pasture infectivity and thus, the worm burden of grazing animals are lowered, especially in young lambs. Research has been conducted throughout the world covering many different climates and management systems. An Australian parasite model showed that if the fungus performs efficiently (> or =90% reduction in worm burden) for 2 or 3 mo, it should contribute significantly to a reduction in the number of dead lambs otherwise occurring when managed only by anthelmintic treatment and grazing management. Feeding or field trials have clearly demonstrated that dosing with a few hundred thousand spores per kilogram of live BW not only reduced the number of infective larvae but also increased the BW of the lambs compared with controls not given fungus. Initial Australian work with feeding spores by means of a block formulation or a slow-release device has shown some promise, but further work is needed to fully develop these delivery systems. In tropical Malaysia, small paddock trials and field studies resulted in significant improvements, in terms of lower worm burdens and increased live BW, when feeding half a million spores daily to grazing lambs. Additional benefits have been observed when the fungus is employed in combination with a fast rotational grazing system. Research has also demonstrated that spores can be delivered in slightly moist feed block material, but only if such blocks are consumed rapidly, because of their very short shelf life. In the northern, temperate Danish climate it has been demonstrated that daily feeding of half a million spores per kilogram of live BW can lead to significant production benefits, with increased live BW gain in fungus-exposed animals. Biological control of parasitic nematodes in sheep seems to hold promise for the future, but to be able to assist producers, the optimal delivery system needs to be refined and further developed. In addition, more work will be needed to define the best use of this technology in different geographic regions.

Matched MeSH terms: Feces/microbiology
Fulltext Blue poo: impact of gut transit time on the gut microbiome using a novel marker

Asnicar F, Leeming ER, Dimidi E, Mazidi M, Franks PW, Al Khatib H, et al.

Gut, 2021 09;70(9):1665-1674.
PMID: 33722860 DOI: 10.1136/gutjnl-2020-323877

BACKGROUND AND AIMS: Gut transit time is a key modulator of host-microbiome interactions, yet this is often overlooked, partly because reliable methods are typically expensive or burdensome. The aim of this single-arm, single-blinded intervention study is to assess (1) the relationship between gut transit time and the human gut microbiome, and (2) the utility of the 'blue dye' method as an inexpensive and scalable technique to measure transit time.
METHODS: We assessed interactions between the taxonomic and functional potential profiles of the gut microbiome (profiled via shotgun metagenomic sequencing), gut transit time (measured via the blue dye method), cardiometabolic health and diet in 863 healthy individuals from the PREDICT 1 study.
RESULTS: We found that gut microbiome taxonomic composition can accurately discriminate between gut transit time classes (0.82 area under the receiver operating characteristic curve) and longer gut transit time is linked with specific microbial species such as Akkermansia muciniphila, Bacteroides spp and Alistipes spp (false discovery rate-adjusted p values <0.01). The blue dye measure of gut transit time had the strongest association with the gut microbiome over typical transit time proxies such as stool consistency and frequency.
CONCLUSIONS: Gut transit time, measured via the blue dye method, is a more informative marker of gut microbiome function than traditional measures of stool consistency and frequency. The blue dye method can be applied in large-scale epidemiological studies to advance diet-microbiome-health research. Clinical trial registry website https://clinicaltrials.gov/ct2/show/NCT03479866 and trial number NCT03479866.

Matched MeSH terms: Feces/microbiology

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