Copro-molecular identification of intestinal nematode infections in a rural community in East Malaysia.

Soil-transmitted helminth (STH) infections are the most common intestinal parasitic infections of medical importance in humans. The infections are widely distributed throughout tropical and subtropical countries, particularly among disadvantaged and underprivileged communities. In Malaysia, STH infections is highly prevalent, especially among the indigenous groups. However, species identification through molecular studies is still lacking. Using molecular techniques, this study was conducted to identify STH species that infect humans. Faecal samples were collected from three Iban tribal longhouses in the rural area of Sarikei Division, Sarawak. Positive faecal samples by microscopy were subjected to PCR and sequenced for species identification. Based on the microscopy examination, the most prevalent STH infection was Ascaris, followed by hookworm and Trichuris. All microscopy-positive samples were then PCR-amplified. The amplicons were sequenced, aligned, and compared with other sequences in the GenBank database. The results showed that Ascaris lumbricoides was the predominant STH species, followed by Trichuris trichiura, Ancylostoma duodenale, and Necator americanus. Our findings were contradictory to past studies, as we showed that A. duodenale was more abundant than N. americanus. Socioeconomic improvement and health education programs should be included in the management and prevention of public health strategies against STH.

Copro-molecular study of Entamoeba infection among the indigenous community in Malaysia: A first report on the species-specific prevalence of Entamoeba in dogs.

BACKGROUND: Entamoeba is a free-living protozoan parasitic species that infect a variety of hosts. In humans, Entamoeba histolytica is the causative agent of amoebiasis. Entamoeba species has also been reported in dogs. However, little is known about the molecular epidemiology and the specific species of this parasite in dogs globally, including Malaysia. As dogs are important companion animals for the indigenous community, and close contact with dogs is part of the natural living conditions for this community, this study aims to determine the prevalence and molecular epidemiology of Entamoeba species in human and dogs in Malaysia. METHOD: The presence of Entamoeba species was examined in 504 fresh fecal samples, collected randomly from 411 humans and 93 dogs using microscopy and polymerase chain reaction (PCR) amplifying 16 s ribosomal RNA (rRNA). Data was analyzed using appropriate statistical analysis. RESULTS: The microscopy data showed an overall occurrence of Entamoeba species of 26.3% (108/411) and 36.6% (34/93) in humans and dogs respectively. In humans, the most common species was a single infection of E. dispar (26.5%; 13/49), followed by E. histolytica and E. moshkovskii, (20.4% for each species respectively). Double infection of E. dispar + E. moshkovskii was detected at 10.2%, followed by E. dispar + E. histolytica (8.2%) and E. moshkovskii and E. histolytica (6.1%). 8.2% of the samples had triple infection with all three species. In animals, E. moshkovskii (46.7%) was the most common species detected, followed by E. histolytica, and E. dispar, at 20.0% and 13.3% respectively. Double infection with E. moshkovskii + E. histolytica and a triple infection were found in 2 samples (13.3%) and 1 (6.7%) sample respectively. Risk factor analysis showed that members of the community who used untreated water were more prone to be infected with Entamoeba. CONCLUSION: This study provides information on the species-specific occurrence of Entamoeba infection, the potential risk factors and their zoonotic potential to humans. This is the first report to describe the molecular occurrence of Entamoeba species in dogs in Malaysia. The presence of pathogenic Entamoeba species implies that dogs could be a reservoir or mechanical host for human amoebiasis. Further studies need to be conducted to better understand the transmission dynamics and public health significance of Entamoeba species in human and animal hosts.

Molecular characterization of Trichuris species isolated from humans, dogs and cats in a rural community in Peninsular Malaysia.

Trichuris trichiura (whipworm) are soil-transmitted helminths (STHs) that causing trichuriasis in human. Trichuris vulpis, a canine whipworm has also been reported occasionally in humans. However, an overlapping dimension in the morphology and due to limited external characters between both species may lead to the potential for misidentification. Although there has been an extensive study on the distribution of whipworm in both human and animal hosts, little is known about the molecular epidemiology of Trichuris species in both hosts. To investigate to characterize the whipworm species and to determine the genetic relationship between species infecting both humans and animals, we sequenced the small subunit ribosomal RNA (SSU rRNA) regions of Trichuris egg isolated from humans, dogs and cats in a rural community in Malaysia. A total of 524 fresh fecal samples were collected from humans and animals. The overall prevalence of Trichuris was 59.9% as determined by microscopy examination. The molecular analysis showed that 98.7% were identified as T. trichiura in the human fecal sample. Interestingly, 1.3% were identified as T. vulpis. As for animal fecal sample, 56.8% and 43.2% were identified as T. trichiura and T. vulpis, respectively. Phylogenetic and sequence analysis demonstrated that T. trichiura isolates were genetically distinct from T. vulpis isolates from both hosts. This finding implies that companion animals can be a reservoir and mechanical transmitter for T. trichiura infection in human and also highlighting the possible zoonotic potential of T. vulpis. This finding may also suggest that cross-transmission between humans and animal hosts in sympatric setting may be a source of infection in both hosts. More studies are needed to better understand the transmission dynamic and public health significance of Trichuris infection in both hosts.

Molecular Diagnosis of Microsporidia among Immunocompromised Patients in Kuala Lumpur, Malaysia.

Microsporidia are obligate intracellular parasitic fungi causing chronic diarrhea, particularly among immunocompromised patients. The current method used for diagnosis is based on conventional microscopy, which does not differentiate parasites at the species level. The present study was carried out to identify microsporidian species in immunocompromised patients. From March 2016 to March 2017, a total of 289 archived stool samples were examined microscopically for microsporidian spores using Gram-chromotrope Kinyoun (GCK) stain. Positive stool samples by microscopy were subjected to polymerase chain reaction and sequencing for species identification. Based on microscopy examination, the overall prevalence of microsporidian infections was 32.2% (93/289; 95% CI = 27.1-37.8). Of these stool samples, 45 samples were successfully amplified and confirmed as Enterocytozoon bieneusi. No Encephalitozoon intestinalis was detected. Accurate identification of species might help clinicians to decide appropriate management strategies as dissemination risks and treatment response vary for different species, hence improving the management of microsporidian infections.