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Intestinal parasitic infections and risk analysis among urban refugees in the Klang Valley, Malaysia

Mohd Hanapi IR, Behnke JM, Sahimin N, Saifulazmi NF, Golam Mohammad Khan ASJ, Abdul Mutalib RNS, et al.

Trans R Soc Trop Med Hyg, 2024 Jan 02;118(1):18-32.
PMID: 37497742 DOI: 10.1093/trstmh/trad047

BACKGROUND: Global studies show intestinal parasitic infections (IPIs) have been introduced and spread with refugee inflows from low to high socio-economic countries. However, there is relatively limited information on the prevalence of infections among the community.
METHODS: A 2-year cross-sectional study was conducted to determine the prevalence and associated risk factors for infections among urban refugees in the Klang Valley, Malaysia. A total of 418 faecal samples were collected and examined by microscopy.
RESULTS: Faecal screening revealed moderate levels (32.3%) of infections in the community. Three nematode (Ascaris lumbricoides, Trichuris trichiura and hookworm) and three protozoan species (Entamoeba, Giardia and Cryptosporidium) were recorded, with the highest prevalence being A. lumbricoides (20.6%) followed by T. trichiura (10.3%), while other infections were <5%. Statistical analysis found that young males with less education were more likely to be infected with helminths. Additionally, living near waste disposal sites, the presence of stray animals, eating with bare hands, bare footedness, poor handwashing practices and no anthelmintic treatment constituted significant risk factors for helminth infections. Protozoan infections were linked to drinking tap water or from water dispensers and poor handwashing practices.
CONCLUSIONS: These findings emphasize the importance of health education in addition to introduction of biannual anthelmintic treatment to promote community health and well-being.

Matched MeSH terms: Cryptosporidiosis*
Effect of nitazoxanide on diarrhea: A systematic review and network meta-analysis of randomized controlled trials

Hashan MR, Elhusseiny KM, Huu-Hoai L, Tieu TM, Low SK, Minh LHN, et al.

Acta Trop, 2020 Oct;210:105603.
PMID: 32598920 DOI: 10.1016/j.actatropica.2020.105603

We aimed to systematically review evidence pertaining to the safety and efficacy of nitazoxanide in treating infectious diarrhea. On September 21, 2017, we identified relevant studies using 12 databases. The estimates of the included studies were pooled as a risk ratio (RR). We conducted a network and pairwise random-effects meta-analysis for both direct and indirect comparisons of different organisms that are known to cause diarrhea. The primary and secondary analysis outcomes were clinical response until cessation of illness, parasitological response and adverse events. We included 18 studies in our analysis. In cryptosporidiosis, the overall estimate favored nitazoxanide in its clinical response in comparison with placebo RR 1.46 [95% CI 1.22-1.74; P-value <0.0001]. Network meta-analysis among patients with Giardia intestinalis showed an increase in the probability of diarrheal cessation and parasitological responses in comparison with placebo, RR 1.69 [95% CI 1.08-2.64, P-score 0.27] and RR 2.91 [95% CI 1.72-4.91, P-score 0.55] respectively. In Clostridium difficile infection, the network meta-analysis revealed a non-significantly superior clinical response effect of nitazoxanide to metronidazole 31 days after treatment RR 1.21 [95% CI 0.87-1.69, P-score 0.26]. In Entamoeba histolytica, the overall estimate significantly favored nitazoxanide in parasitological response with placebo RR 1.80 [95% CI 1.35-2.40, P-value < 0.001]. We highlighted the effectiveness of nitazoxanide in the cessation of diarrhea caused by Cryptosporidium, Giardia intestinalis and Entamoeba histolytica infection. We also found significant superiority of NTZ to metronidazole in improving the clinical response to G. intestinalis, thus it may be a suitable candidate for treating infection-induced diarrhea. To prove the superiority of NTZ during a C. difficile infection may warrant a larger-scale clinical trial since its superiority was deemed insignificant. We recommend nitazoxanide as an appropriate option for treating infectious diarrhea.

Matched MeSH terms: Cryptosporidiosis/drug therapy
Prevalence of Cryptosporidium spp. infection among children admitted to Hospital Tengku Ampuan Afzan

Asady A, Ismail S, Marsitah AJ, Pakeer O

Med J Malaysia, 2019 12;74(6):468-471.
PMID: 31929470

INTRODUCTION: Cryptosporidium spp. is identified as an important cause of diarrhoeal morbidity and mortality worldwide particularly in children below five years of age and immunocompromised individuals. Infections are present among cattle and humans. Until now, there is no report on its prevalence in humans in Kuantan. The aim of this study is to record the prevalence and associated risk factors of Cryptosporidium spp. infection among children admitted to Hospital Tengku Ampuan Afzan (HTAA), Kuantan.
METHODS: This is a cross-sectional study conducted among children admitted to the Paediatrics ward in HTAA between December 2017 and May 2018. Faecal samples were examined using wet smear and Modified Ziehl-Neelsen (MZN) staining techniques. Data on demography and hygiene practices was collected using a pretested questionnaire, and analysed using SPSS version 22.
RESULTS: One hundred thirty five children (95.6% were of Malay ethnicity) were included in the study. The overall prevalence of Cryptosporidium spp. infection was 25.2%. The prevalence was slightly higher in females (28.1%) than males (23.1%). The prevalence was higher than expected. This study showed that some risk factors namely children's age and trash disposal methods were significantly associated with Cryptosporidium spp. infection (p<0.05).
CONCLUSION: Higher prevalence could possibly be due to an outbreak of this infection or until now undetected.

Matched MeSH terms: Cryptosporidiosis/microbiology; Cryptosporidiosis/epidemiology*
Occurrence of zoonotic Cryptosporidium and Giardia duodenalis species/genotypes in urban rodents

Tan TK, Low VL, Ng WH, Ibrahim J, Wang D, Tan CH, et al.

Parasitol Int, 2019 Apr;69:110-113.
PMID: 30590124 DOI: 10.1016/j.parint.2018.12.007

This report describes the detection of zoonotic Cryptosporidium muris, C. parvum subgenotype IIa and Giardia duodenalis genotype B in urban rodents in Malaysia. A rare occurrence of C. meleagridis was also reported suggesting a role of rodents in mechanical transmission of this pathogen. Utilization of DNA sequencing and subtyping analysis confirmed the presence of zoonotic C. parvum subtypes IIaA17G2R1 and IIaA16G3R1 for the first time in rodents.

Matched MeSH terms: Cryptosporidiosis
Molecular diagnosis and genetic diversity of Cryptosporidium spp. in exotic birds of southwest of Iran

Jalas M, Tavalla M

Trop Biomed, 2018 Dec 01;35(4):944-950.
PMID: 33601843

Cryptosporidium parasites can infect a wide range of vertebrate hosts including reptiles, mammals, and birds. Due to the zoonotic nature of cryptosporidiosis and its close contact with exotic birds and humans, the present study aimed to determine the prevalence and genetic diversity of Cryptosporidium spp. in exotic birds of southwest of Iran, by the staining and molecular methods. In the present research, 369 stool specimens were randomly collected from exotic birds and stained by modified acid-fast stain using Ziehl-Neelsen method. The slides were examined using light microscopy at a magnification of 100X. Then, the extracted DNA was amplified using the PCR method. Finally, all genotypes and positive samples from PCR assay were sequenced by Bioneer Company (Daejeon, South Korea). Among 369 stool specimens, 25 and 27 cases were found to be positive for Cryptosporidium spp. by the modified Ziehl-Neelsen staining and the PCR methods, respectively. Based on the genotyping, C. avian genotype III and C. meleagridis were detected in 25 and 2 stool samples, respectively. The results revealed a relatively high prevalence of Cryptosporidium spp. in exotic birds in the southwest of Iran. Due to the zoonotic nature of C. meleagridis, these exotic birds can be a significant source of cryptosporidiosis. It is important that high-risk people, including immune-deficient patients, receive correct information about the risk of indirect and direct contact with infected exotic birds.

Matched MeSH terms: Cryptosporidiosis
Parasites in soil samples from Signy island, Antarctica

Lim PKC, Lee XC, Mohd Nazmi NMA, Tang YY, Wong SF, Mak JW, et al.

Trop Biomed, 2018 Dec 01;35(4):1007-1016.
PMID: 33601848

Studies on parasite populations in Antarctic soils are scarce and thus little is known about the threat of these parasites towards either the natural fauna or human visitors. However, human presence in Antarctica, mainly through research and tourism, keeps increasing over time, potentially exposing visitors to zoonotic infections from Antarctic wildlife and environment. Most available literature to date has focused on faecal samples from Antarctic vertebrates. Therefore, this study addressed the possible presence of parasites in Antarctic soil that may be infectious to humans. Soil samples were obtained from five locations on Signy Island (South Orkney Islands, maritime Antarctic), namely North Point and Gourlay Peninsula (penguin rookeries), Pumphouse (relic coal-powered pump house), Jane Col (barren high altitude fellfield) and Berntsen Point (low altitude vegetated fellfield close to current research station). Approximately 10% of the soil samples (14/135) from 3 out of the 5 study sites had parasites which included Diphyllobotridae spp. eggs, Cryptosporidium sp., an apicomplexan protozoa (gregarine), Toxoplasma gondii, helminths (a cestode, Tetrabothrius sp., and a nematode larva) and mites. The presence of parasites in the 3 sites are most likely due to the presence of animal and human activities as two of these sites are penguin rookeries (North Point and Gourlay Peninsula) while the third site (Pumphouse Lake) has human activity. While some of the parasite species found in the soil samples appear to be distinctive, there were also parasites such as Cryptosporidium and Toxoplasma gondii that have a global distribution and are potentially pathogenic.

Matched MeSH terms: Cryptosporidiosis
In silico identification and validation of a novel hypothetical protein in Cryptosporidium hominis and virtual screening of inhibitors as therapeutics

Shrivastava AK, Kumar S, Sahu PS, Mahapatra RK

Parasitol Res, 2017 May;116(5):1533-1544.
PMID: 28389892 DOI: 10.1007/s00436-017-5430-1

Computational approaches to predict structure/function and other biological characteristics of proteins are becoming more common in comparison to the traditional methods in drug discovery. Cryptosporidiosis is a major zoonotic diarrheal disease particularly in children, which is caused primarily by Cryptosporidium hominis and Cryptosporidium parvum. Currently, there are no vaccines for cryptosporidiosis and recommended drugs are ineffective. With the availability of complete genome sequence of C. hominis, new targets have been recognized for the development of effective and better drugs and/or vaccines. We identified a unique hypothetical protein (TU502HP) in the C. hominis genome from the CryptoDB database. A three-dimensional model of the protein was generated using the Iterative Threading ASSEmbly Refinement server through an iterative threading method. Functional annotation and phylogenetic study of TU502HP protein revealed similarity with human transportin 3. The model is further subjected to a virtual screening study form the ZINC database compound library using the Dock Blaster server. A docking study through AutoDock software reported N-(3-chlorobenzyl)ethane-1,2-diamine as the best inhibitor in terms of docking score and binding energy. The reliability of the binding mode of the inhibitor is confirmed by a complex molecular dynamics simulation study using GROMACS software for 10 ns in the water environment. Furthermore, antigenic determinants of the protein were determined with the help of DNASTAR software. Our findings report a great potential in order to provide insights in the development of new drug(s) or vaccine(s) for treatment and prophylaxis of cryptosporidiosis among humans and animals.

Matched MeSH terms: Cryptosporidiosis/diagnosis; Cryptosporidiosis/drug therapy*
Fulltext Revisiting the global problem of cryptosporidiosis and recommendations

Shrivastava AK, Kumar S, Smith WA, Sahu PS

Trop Parasitol, 2017 Jan-Jun;7(1):8-17.
PMID: 28459010 DOI: 10.4103/2229-5070.202290

Cryptosporidiosis is a gastrointestinal illness caused by the protozoan parasite Cryptosporidium species, which is a leading cause of diarrhea in a variety of vertebrate hosts. The primary mode of transmission is through oral routes; infections spread with the ingestion of oocysts by susceptible animals or humans. In humans, Cryptosporidium infections are commonly found in children and immunocompromised individuals. The small intestine is the most common primary site of infection in humans while extraintestinal cryptosporidiosis occurs in immunocompromised individuals affecting the biliary tract, lungs, or pancreas. Both innate and adaptive immune responses play a critical role in parasite clearance as evident from studies with experimental infection in mice. However, the cellular immune responses induced during human infections are poorly understood. In this article, we review the currently available information with regard to epidemiology, diagnosis, therapeutic interventions, and strategies being used to control cryptosporidiosis infection. Since cryptosporidiosis may spread through zoonotic mode, we emphasis on more epidemiological surveillance-based studies in developing countries with poor sanitation and hygiene. These epidemiological surveys must incorporate fecal source tracking measures to identify animal and human populations contributing significantly to the fecal burden in the community, as mitigation measures differ by host type.

Matched MeSH terms: Cryptosporidiosis
Fulltext Detection of Cryptosporidium and Cyclospora Oocysts from Environmental Water for Drinking and Recreational Activities in Sarawak, Malaysia

Bilung LM, Tahar AS, Yunos NE, Apun K, Lim YA, Nillian E, et al.

Biomed Res Int, 2017;2017:4636420.
PMID: 29234679 DOI: 10.1155/2017/4636420

Cryptosporidiosis and cyclosporiasis are caused by waterborne coccidian protozoan parasites of the genera Cryptosporidium and Cyclospora, respectively. This study was conducted to detect Cryptosporidium and Cyclospora oocysts from environmental water abstracted by drinking water treatment plants and recreational activities in Sarawak, Malaysia. Water samples (12 each) were collected from Sungai Sarawak Kanan in Bau and Sungai Sarawak Kiri in Batu Kitang, respectively. In addition, 6 water samples each were collected from Ranchan Recreational Park and UNIMAS Lake at Universiti Malaysia Sarawak, Kota Samarahan, respectively. Water physicochemical parameters were also recorded. All samples were concentrated by the iron sulfate flocculation method followed by the sucrose floatation technique. Cryptosporidium and Cyclospora were detected by modified Ziehl-Neelsen technique. Correlation of the parasites distribution with water physicochemical parameters was analysed using bivariate Pearson correlation. Based on the 24 total samples of environmental water abstracted by drinking water treatment plants, all the samples (24/24; 100%) were positive with Cryptosporidium, and only 2 samples (2/24; 8.33%) were positive with Cyclospora. Based on the 12 total samples of water for recreational activities, 4 samples (4/12; 33%) were positive with Cryptosporidium, while 2 samples (2/12; 17%) were positive with Cyclospora. Cryptosporidium oocysts were negatively correlated with dissolved oxygen (DO).

Matched MeSH terms: Cryptosporidiosis/parasitology; Cryptosporidiosis/transmission
Fulltext Identification of Cryptosporidium from Dairy Cattle in Pahang, Malaysia

Hisamuddin NH, Hashim N, Soffian SN, Amin MH, Wahab RA, Mohammad M, et al.

Korean J Parasitol, 2016 Apr;54(2):197-200.
PMID: 27180579 DOI: 10.3347/kjp.2016.54.2.197

Cryptosporidium, a protozoan parasite, can cause cryptosporidiosis which is a gastrointestinal disease that can infect humans and livestock. Cattle are the most common livestock that can be infected with this protozoan. This study was carried out to determine the prevalence of Cryptosporidium infection in cattle in Kuantan, Pahang, Malaysia and to find out the association between the occurrence of infection and 3 different ages of cattle (calves less than 1 year, yearling, and adult cattle). The samples were processed by using formol-ether concentration technique and stained by modified Ziehl Neelsen. The results showed that 15.9% (24/151) of cattle were positive for Cryptosporidium oocysts. The occurrence of Cryptosporidium in calves less than 1 year was the highest with the percentage of 20.0% (11/55) followed by yearling and adult cattle, with the percentage occurrence of 15.6 % (7/45) and 11.8% (6/51), respectively. There was no significant association between the occurrence and age of cattle and presence of diarrhea. Good management practices and proper hygiene management must be taken in order to reduce the infection. It is highly important to control the infection since infected cattle may serve as potential reservoirs of the infection to other animals and humans, especially animal handlers.

Matched MeSH terms: Cryptosporidiosis/epidemiology*; Cryptosporidiosis/parasitology
Molecular analysis of Cryptosporidium from cattle from five states of Peninsular Malaysia

Yap NJ, Koehler AV, Ebner J, Tan TK, Lim YA, Gasser RB

Mol Cell Probes, 2016 Feb;30(1):39-43.
PMID: 26775614 DOI: 10.1016/j.mcp.2016.01.002

Despite the importance of the cattle industry in Malaysia, there are very few studies of the diversity and public health significance of bovine cryptosporidiosis in this country. In the present study, we used a PCR-based approach to detect and genetically characterize Cryptosporidium DNA in faecal samples from a cohort of 215 asymptomatic cattle (of different ages) from six farms from five states of Peninsular Malaysia. Cattle on four of the six farms were test-positive for Cryptosporidium, with an overall prevalence of 3.2%. Cryptosporidium bovis and Cryptosporidium ryanae were detected in two (0.9%) and five (2.3%) samples tested; this low prevalence likely relates to the age of the cattle tested, as most (73%) of the samples tested originated from cattle that were ≥2 years of age. Future studies should investigate the zoonotic potential of Cryptosporidium in pre-weaned and weaned calves in rural communities of Malaysia.

Matched MeSH terms: Cryptosporidiosis/diagnosis; Cryptosporidiosis/parasitology*
Fulltext Molecular epidemiology of Cryptosporidium in HIV/AIDS patients in Malaysia

Asma I, Sim BL, Brent RD, Johari S, Yvonne Lim AL

Trop Biomed, 2015 Jun;32(2):310-22.
PMID: 26691260 MyJurnal

Cryptosporidiosis is a particular concern in immunocompromised individuals where symptoms may be severe. The aim of this study was to examine the epidemiological and molecular characteristics of Cryptosporidium infections in HIV/AIDS patients in Malaysia in order to identify risk factors and facilitate control measures. A modified Ziehl-Neelsen acid fast staining method was used to test for the presence of Cryptosporidium oocysts in the stools of 346 HIV/AIDS patients in Malaysia. Standard coproscopical methods were used to identify infections with other protozoan or helminths parasites. To identify the species of Cryptosporidium, DNA was extracted and nested-PCR was used to amplify a portion of the SSU rRNA gene. A total of 43 (12.4%) HIV-infected patients were found to be infected with Cryptosporidium spp. Of the 43 Cryptosporidium-positive HIV patients, 10 (23.3%) also harboured other protozoa, and 15 (34.9%) had both protozoa and helminths. The highest rates of cryptosporidiosis were found in adult males of Malay background, intravenous drug users, and those with low CD4 T cell counts (i.e., < 200 cells/mm3). Most were asymptomatic and had concurrent opportunistic infections mainly with Mycobacterium tuberculosis. DNA sequence analysis of 32 Cryptosporidium isolates identified C. parvum (84.3%), C. hominis (6.3%), C. meleagridis (6.3%), and C. felis (3.1%). The results of the present study revealed a high prevalence of Cryptosporidium infection in hospitalized HIV/AIDS patients. The results also confirmed the potential significance of zoonotic transmission of C. parvum in HIV infected patients, as it was the predominant species found in this study. However, these patients were found to be susceptible to a wide range of Cryptosporidium species. Epidemiological and molecular characterization of Cryptosporidium isolates provides clinicians and researchers with further information regarding the origin of the infection, and may enhance treatment and control strategies.

Matched MeSH terms: Cryptosporidiosis/epidemiology*; Cryptosporidiosis/parasitology*
An improved stool concentration procedure for the detection of Cryptosporidium oocysts in Orang Asli stool samples

Salleh FM, Moktar N, Yasin AM, Al-Mekhlafi HM, Anuar TS

J Microbiol Methods, 2014 Nov;106:143-145.
PMID: 25193442 DOI: 10.1016/j.mimet.2014.08.019

To improve the stool concentration procedure, we modified different steps of the standard formalin-ether concentration technique and evaluated these modifications by examining stool samples collected in the field. Seven samples were found positive by the modified formalin-ether concentration technique (M-FECT). Therefore, the M-FECT procedure provides enhanced detection of Cryptosporidium oocysts.

Matched MeSH terms: Cryptosporidiosis/diagnosis*; Cryptosporidiosis/microbiology
Fulltext Elongation factor-1α is a novel protein associated with host cell invasion and a potential protective antigen of Cryptosporidium parvum

Matsubayashi M, Teramoto-Kimata I, Uni S, Lillehoj HS, Matsuda H, Furuya M, et al.

J Biol Chem, 2013 Nov 22;288(47):34111-34120.
PMID: 24085304 DOI: 10.1074/jbc.M113.515544

The phylum Apicomplexa comprises obligate intracellular parasites that infect vertebrates. All invasive forms of Apicomplexa possess an apical complex, a unique assembly of organelles localized to the anterior end of the cell and involved in host cell invasion. Previously, we generated a chicken monoclonal antibody (mAb), 6D-12-G10, with specificity for an antigen located in the apical cytoskeleton of Eimeria acervulina sporozoites. This antigen was highly conserved among Apicomplexan parasites, including other Eimeria spp., Toxoplasma, Neospora, and Cryptosporidium. In the present study, we identified the apical cytoskeletal antigen of Cryptosporidium parvum (C. parvum) and further characterized this antigen in C. parvum to assess its potential as a target molecule against cryptosporidiosis. Indirect immunofluorescence demonstrated that the reactivity of 6D-12-G10 with C. parvum sporozoites was similar to those of anti-β- and anti-γ-tubulins antibodies. Immunoelectron microscopy with the 6D-12-G10 mAb detected the antigen both on the sporozoite surface and underneath the inner membrane at the apical region of zoites. The 6D-12-G10 mAb significantly inhibited in vitro host cell invasion by C. parvum. MALDI-TOF/MS and LC-MS/MS analysis of tryptic peptides revealed that the mAb 6D-12-G10 target antigen was elongation factor-1α (EF-1α). These results indicate that C. parvum EF-1α plays an essential role in mediating host cell entry by the parasite and, as such, could be a candidate vaccine antigen against cryptosporidiosis.

Matched MeSH terms: Cryptosporidiosis/genetics; Cryptosporidiosis/immunology; Cryptosporidiosis/metabolism; Cryptosporidiosis/prevention & control
First molecular characterization of Cryptosporidium in Yemen

Alyousefi NA, Mahdy MA, Lim YA, Xiao L, Mahmud R

Parasitology, 2013 May;140(6):729-34.
PMID: 23369243 DOI: 10.1017/S0031182012001953

Cryptosporidium is a protozoan parasite of humans and animals and has a worldwide distribution. The parasite has a unique epidemiology in Middle Eastern countries where the IId subtype family of Cryptosporidium parvum dominates. However, there has been no information on Cryptosporidium species in Yemen. Thus, this study was conducted in Yemen to examine the distribution of Cryptosporidium species and subtype families. Fecal samples were collected from 335 patients who attended hospitals in Sana'a city. Cryptosporidium species were determined by PCR and sequence analysis of the 18 s rRNA gene. Cryptosporidium parvum and C. hominis subtypes were identified based on sequence analysis of the 60 kDa glycoprotein (gp60) gene. Out of 335 samples, 33 (9.9%) were positive for Cryptosporidium. Of them, 97% were identified as C. parvum whilst 1 case (3%) was caused by C. hominis. All 7 C. parvum isolates subtyped belonged to the IIaA15G2R1 subtype. The common occurrence of the zoonotic IIa subtype family of C. parvum highlights the potential occurrence of zoonotic transmission of cryptosporidiosis in Yemen. However, this postulation needs confirmation with future molecular epidemiological studies of cryptosporidiosis in both humans and animals in Yemen.

Matched MeSH terms: Cryptosporidiosis/epidemiology; Cryptosporidiosis/parasitology*
Fulltext High diversity of Cryptosporidium subgenotypes identified in Malaysian HIV/AIDS individuals targeting gp60 gene

Iqbal A, Lim YA, Surin J, Sim BL

PLoS One, 2012;7(2):e31139.
PMID: 22347442 DOI: 10.1371/journal.pone.0031139

Currently, there is a lack of vital information in the genetic makeup of Cryptosporidium especially in developing countries. The present study aimed at determining the genotypes and subgenotypes of Cryptosporidium in hospitalized Malaysian human immunodeficiency virus (HIV) positive patients.

Matched MeSH terms: Cryptosporidiosis/classification
First genetic classification of Cryptosporidium and Giardia from HIV/AIDS patients in Malaysia

Lim YA, Iqbal A, Surin J, Sim BL, Jex AR, Nolan MJ, et al.

Infect Genet Evol, 2011 Jul;11(5):968-74.
PMID: 21439404 DOI: 10.1016/j.meegid.2011.03.007

Given the HIV epidemic in Malaysia, genetic information on opportunistic pathogens, such as Cryptosporidium and Giardia, in HIV/AIDS patients is pivotal to enhance our understanding of epidemiology, patient care, management and disease surveillance. In the present study, 122 faecal samples from HIV/AIDS patients were examined for the presence of Cryptosporidium oocysts and Giardia cysts using a conventional coproscopic approach. Such oocysts and cysts were detected in 22.1% and 5.7% of the 122 faecal samples, respectively. Genomic DNAs from selected samples were tested in a nested-PCR, targeting regions of the small subunit (SSU) of nuclear ribosomal RNA and the 60kDa glycoprotein (gp60) genes (for Cryptosporidium), and the triose-phosphate isomerase (tpi) gene (for Giardia), followed by direct sequencing. The sequencing of amplicons derived from SSU revealed that Cryptosporidium parvum was the most frequently detected species (64% of 25 samples tested), followed by C. hominis (24%), C. meleagridis (8%) and C. felis (4%). Sequencing of a region of gp60 identified C. parvum subgenotype IIdA15G2R1 and C. hominis subgenotypes IaA14R1, IbA10G2R2, IdA15R2, IeA11G2T3R1 and IfA11G1R2. Sequencing of amplicons derived from tpi revealed G. duodenalis assemblage A, which is of zoonotic importance. This is the first report of C. hominis, C. meleagridis and C. felis from Malaysian HIV/AIDS patients. Future work should focus on an extensive analysis of Cryptosporidium and Giardia in such patients as well as in domestic and wild animals, in order to improve the understanding of transmission patterns and dynamics in Malaysia. It would also be particularly interesting to establish the relationship among clinical manifestation, CD4 cell counts and genotypes/subgenotypes of Cryptosporidium and Giardia in HIV/AIDS patients. Such insights would assist in a better management of clinical disease in immuno-deficient patients as well as improved preventive and control strategies.

Matched MeSH terms: Cryptosporidiosis/complications; Cryptosporidiosis/epidemiology; Cryptosporidiosis/parasitology*
Molecular identification of Cryptosporidium parvum from avian hosts

Quah JX, Ambu S, Lim YA, Mahdy MA, Mak JW

Parasitology, 2011 Apr;138(5):573-7.
PMID: 21232175 DOI: 10.1017/S0031182010001691

Cryptosporidium species are protozoan parasites that infect humans and a wide variety of animals. This study was aimed at identifying Cryptosporidium species and genotypes isolated from avian hosts. A total of 90 samples from 37 different species of birds were collected throughout a 3-month period from April 2008 to June 2008 in the National Zoo of Kuala Lumpur, Malaysia. Prior to molecular characterization, all samples were screened for Cryptosporidium using a modified Ziehl-Neelsen staining technique. Subsequently samples were analysed with nested-PCR targeting the partial SSU rRNA gene. Amplicons were sequenced in both directions and used for phylogenetic analysis using Neighbour-Joining and Maximum Parsimony methods. Although 9 (10%) samples were positive for Cryptosporidium via microscopy, 8 (8.9%) produced amplicons using nested PCR. Phylogenetic trees identified all the isolates as Cryptosporidium parvum. Although C. parvum has not been reported to cause infection in birds, and the role of birds in this study was postulated mainly as mechanical transporters, these present findings highlight the significant public health risk posed by birds that harbour the zoonotic species of Cryptosporidium.

Matched MeSH terms: Cryptosporidiosis/epidemiology; Cryptosporidiosis/parasitology
Fulltext Childhood Cryptosporidium infection among aboriginal communities in Peninsular Malaysia

Al-Mekhlafi HM, Mahdy MA, 'Azlin MY, Fatmah MS, Norhayati M

Ann Trop Med Parasitol, 2011 Mar;105(2):135-43.
PMID: 21396250 DOI: 10.1179/136485911X12899838683368

Cryptosporidium is a coccidian parasite that is prevalent worldwide, some species of which cause morbidity in both immunocompromised and immunocompetent individuals. The prevalence and predictors of Cryptosporidium infection, and its effect on nutritional status, have recently been explored among 276 children (141 boys and 135 girls, aged 2-15 years) in aboriginal (Orang Asli) villages in the Malaysian state of Selangor. Faecal smears were examined by the modified Ziehl-Neelsen staining technique while socio-economic data were collected using a standardized questionnaire. Nutritional status was assessed by anthropometric measurements. Cryptosporidium infection, which was detected in 7.2% of the aboriginal children, was found to be significantly associated with low birthweight (≤2.5 kg), being part of a large household (with more than seven members) and prolonged breast feeding (>2 years). The output of a binary logistic regression confirmed that large household size was a significant predictor of Cryptosporidium infection (giving an odds ratio of 2.15, with a 95% confidence interval of 1.25-5.02). Cryptosporidium infection is clearly a public-health problem among the aboriginal children of Selangor, with person-to-person the most likely mode of transmission.

Matched MeSH terms: Cryptosporidiosis/ethnology*; Cryptosporidiosis/etiology; Cryptosporidiosis/physiopathology
Prevalence of and management factors contributing to Cryptosporidium sp. infection in pre-weaned and post-weaned calves in Johor, Malaysia

Muhid A, Robertson I, Ng J, Ryan U

Exp Parasitol, 2011 Feb;127(2):534-8.
PMID: 21050848 DOI: 10.1016/j.exppara.2010.10.015

A cross-sectional study was carried out to identify species and determine the prevalence of Cryptosporidium sp. shedding in pre-weaned and post-weaned dairy calves and to identify management factors that may be contributing to disease. A total of 240 calf faecal samples were collected from 16 farms in two districts in Johor, Malaysia, and screened by PCR. The overall Cryptosporidium prevalence was 27.1%. The prevalence of Cryptosporidium species in pre-weaned calves was 32.4% for C. parvum, 26.5% for C. bovis, followed by C. andersoni (20.6%), C. ryanae (11.8%) and mixed sp. (8.8%). The prevalence of Cryptosporidium species in post-weaned calves was 35% for C. bovis followed by C. andersoni and C. ryanae (30% each) and mixed sp. (5%). Subtyping analysis of 8 of the 11 C. parvum isolates at the gp60 locus identified five isolates as IIdA15G1, one as IIa18A3R1 and two isolates as IIa17G2R1. Management factors that increased the risk of Cryptosporidium infection included having other cattle farms close by, feeding calves with saleable milk, keeping pre-weaned calves in pens with slatted floors and keeping post-weaned calves in pens with a sand floor.

Matched MeSH terms: Cryptosporidiosis/epidemiology; Cryptosporidiosis/prevention & control

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