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Cor pulmonale: an unusual presentation of tropical eosinophilia

Quah BS, Anuar AK, Rowani MR, Pennie RA

Ann Trop Paediatr, 1997 Mar;17(1):77-81.
PMID: 9176582

Tropical pulmonary eosinophilia (TPE) is considered to be a variant of human filarial infection. The pulmonary manifestations of TPE have been well described. Extra-pulmonary features of the disease, although not commonly seen, have been reported previously. A 9-year-old Malay girl with a history of recurrent cough and wheezing was admitted because of cardiac failure. Physical examination revealed a very sick girl with tachypnoea, central cyanosis, finger clubbing, elevated jugular venous pulse, generalized crackles and rhonchi in the chest, a loud second heart sound and hepatosplenomegaly. A chest radiograph showed cardiomegaly and right pleural effusion. Laboratory investigations revealed hypochromic, microcytic anaemia with persistent blood eosinophilia (absolute eosinophil counts varied from 1.9 to 5.5 x 10(9)/1). The ELISA test for antifilarial IgG antibodies was strongly positive. She responded promptly to treatment with diethylcarbamazine. In summary, this is a patient with TPE who presented with cor pulmonale, probably due to late-stage interstitial pulmonary fibrosis. In order to prevent the long term morbidity of cardiorespiratory disability, the early signs of TPE should be recognized and the infection treated.
Irregular shedding of Blastocystis hominis

Vennila GD, Suresh Kumar G, Khairul Anuar A, Rajah S, Saminathan R, Sivanandan S, et al.

Parasitol Res, 1999 Feb;85(2):162-4.
PMID: 9934969

The shedding pattern of the protozoan parasite, Blastocystis hominis, is investigated in man and in experimental animal infections. The shedding pattern of the vacuolar and cystic forms of Blastocystis hominis in infected individuals have been shown in the present study to be irregular. The study shows that there is marked fluctuation in the shedding of the parasite from day to day, varying from as high as 17 to 0 per x40 microscopic field. The cystic stages when estimated in 8 Blastocystis-infected individuals ranged from as high as 7.4x10(5) cysts per gram of stool to 0. The shedding of cystic and vacuolar forms observed over a period of 20 days in experimentally-infected Wistar rats were not only shown to be irregular but the amount varied from host to host. The study has important diagnostic implications in that the stool samples must be collected more than once from patients showing clinical signs and symptoms to eliminate the cause of it to Blastocystis. The study also shows that there are asymptomatic individuals who pass a large amount of cysts as such individuals should be treated to prevent transmission to others.
Fulltext Cryptosporidiosis among children with acute gastroenteritis in the pediatric ward in the General Hospital, Penang

Ludin CM, Afifi SA, Hasenan N, Maimunah A, Anuar AK

Southeast Asian J. Trop. Med. Public Health, 1991 Jun;22(2):200-2.
PMID: 1948280

Stool samples from 836 cases with diarrhea and acute gastroenteritis from the Pediatric ward, Penang General Hospital, were examined for Cryptosporidium oocysts. A dimethyl sulfoxide modified acid fast technique was used for the identification of the parasites. 36 samples or 4.3% were found to be positive for Cryptosporidium. The prevalence of infection was higher (2.39%) in children with diarrhea and vomiting than in children with acute gastroenteritis alone (0.8%). Stool examination and cultures from the Cryptosporidium positive samples revealed no other parasites, rotavirus or enteropathogenic bacteria. This suggests that Cryptosporidium may be an important agent in the causation of diarrhea in young children. A routine laboratory examination for the detection of Cryptosporidium in the search for causal agents of childhood diarrhea in our environment may, therefore, be significant.
Side Effects of Chronic Methadone Use in Opioid Addiction: A Scoping Review on Hematological Derangements

Khalid K, Anuar A, Ang WC, Jamaluddin R

Curr Drug Saf, 2023;18(2):167-174.
PMID: 35570535 DOI: 10.2174/1574886317666220514154931

BACKGROUND: Per oral methadone maintenance therapy [MMT] was introduced as part of harm reduction strategy to manage rising numbers of HIV infections caused by intravenous use of illicit drugs.
METHOD: We aim to review published articles related to the side effects of long-term methadone therapy, focusing on hematological derangements in human studies published between 1 January 2000 till 31 January 2021.
RESULTS: Our search databases include Web of Science, Scopus, and Medline. Our search yielded 971 articles, of which 55 articles were related to the effects of MMT on various organ systems: cardiovascular [n=12], respiratory [n=1], endocrine [n=10], central nervous system [n=12], neurobehavioral [n=10], gastrointestinal [n=1], and bone [n=1]. There were eight articles specifically related to the hematological side effects of chronic methadone therapy that include [i] immune system hyperactivation, [ii] reduced circulating lymphocytes, and [iii] increased blood viscosity.
CONCLUSION: In view of all foreseeable health risks seen with prolonged methadone therapy, pharmacological modulation is warranted to find a better substitute for managing patients with opioid dependence.
Bibliometric analysis of immigration and environmental degradation: evidence from past decades

Anuar A, Marwan NF, Smith J, Siriyanun S, Sharif A

Environ Sci Pollut Res Int, 2022 Feb;29(9):13729-13741.
PMID: 34599441 DOI: 10.1007/s11356-021-16470-1

The aim of this paper is to examine immigration and environmental degradation using bibliometric analysis. This paper also analyzes sources of publication, authorship, citations, distributions publications and other bibliometric indicators. The study focuses on a total of 1372 articles published from 2000 to 2020. These articles were collected through an automated process from the Scopus database and later analyzed using techniques such as bibliometric indicators analysis, VOSviewer, and Perish or Publish. The research identified 991 articles from varieties of published sources. The topic of immigrants and environmental degradation has been an emerging topic since 1981. Starting in 2000, most of the scholars actively producing an articles pertinent to this topic. Most of the articles were published in journals, and English is the primary language of research. United States is the leading country in contributing the publications. Meanwhile, the most significant fields in which the sources were produced were environmental science, agricultural and biological sciences, arts and humanities and earth and planetary sciences. However, some limitations has been found. It has been suggested for future research, to lengthen this work to other databases, as well as bibliometric analyses of immigration and environmental degradation in developed and developing countries by adding a new keyword such as energy consumption and climate change. This paper aims to assess recent trends in the expansion of academic literature on immigration and environmental degradation using the bibliometric analysis method. Network visualization and bibliometric indicators are used in this paper to present the results.
Fulltext Effect of Temperatures on Drying Kinetics, Extraction Yield, Phenolics, Flavonoids, and Antioxidant Activity of Phaleria macrocarpa (Scheff.) Boerl. (Mahkota Dewa) Fruits

Stephenus FN, Benjamin MAZ, Anuar A, Awang MA

Foods, 2023 Jul 27;12(15).
PMID: 37569127 DOI: 10.3390/foods12152859

Phaleria macrocarpa (Scheff.) Boerl. or 'Mahkota Dewa' is a popular plant found in Malaysia as it is a valuable source of phytochemicals and therapeutic properties. Drying is an essential step in the storage of P. macrocarpa fruits at an industrial level to ensure their availability for a prolonged shelf life as well as preserving their bioactive compounds. Hence, this study evaluates the effect of different temperatures on the drying kinetics, extraction yield, phenolics, flavonoids, and antioxidant activity of P. macrocarpa fruits. The oven-drying process was carried out in this study at temperatures of 40 °C, 50 °C, 60 °C, 70 °C, and 80 °C. Six thin-layer drying models (i.e., Lewis, Page, Henderson and Pabis, two-term exponential, Logarithmic, and Midilli and Kucuk models) were evaluated to study the behaviour of oven-dried P. macrocarpa fruits based on the coefficient of determination (R2), root mean square error (RMSE), and chi-square (χ2). The quality of the oven-dried P. macrocarpa fruits was determined based on their extraction yield, total phenolic content (TPC), total flavonoid content (TFC), and antioxidant activity (2,2-diphenyl-1-picrylhydrazyl) using ultrasound-assisted extraction. The results showed that the time for moisture removal correspondingly increased in the oven-dried P. macrocarpa fruits. Apparently, the Midilli and Kucuk model is the most appropriate model to describe the drying process. The range of effective moisture diffusivity was 1.22 × 10-8 to 4.86 × 10-8 m2/s, and the activation energy was 32.33 kJ/mol. The oven-dried P. macrocarpa fruits resulted in the highest extraction yield (33.99 ± 0.05%), TPC (55.39 ± 0.03 mg GAE/g), TFC (15.47 ± 0.00 mg RE/g), and DPPH inhibition activity (84.49 ± 0.02%) at 60 °C based on the significant difference (p < 0.05). A strong correlation was seen between the antioxidant activity, TPC, and TFC in the oven-dried P. macrocarpa fruits. The current study suggests that the oven-drying method improved the TPC, TFC, and antioxidant activity of the P. macrocarpa fruits, which can be used to produce functional ingredients in foods and nutraceuticals.
The level of knowledge and acceptance towards the COVID-19 vaccine among the community in Johor Bahru, Johor

Mohd Anuar AH, Mohamad Anuar NN, Isa SNI, Bahari M

Med J Malaysia, 2024 Mar;79(Suppl 1):88-95.
PMID: 38555891

INTRODUCTION: Several necessary initiatives have been made to create a readily available vaccine against the Coronavirus Disease 2019 (COVID-19) worldwide pandemic. However, the vaccination program's success is dependent on the population's willingness as well as their knowledge of vaccination. Hence, the present study aimed to assess the level of knowledge and acceptance towards the COVID-19 vaccine among the community in Johor Bahru, Johor.
MATERIALS AND METHODS: A cross-sectional study was conducted using an online survey between February and May 2022, with 423 respondents. The questionnaire consisted of socio-demographic, assessment of knowledge level and acceptance level towards COVID-19 vaccine. The descriptive analysis and non-parametric tests were employed to investigate the study outline objectives.
RESULTS: Of all 423 participants, 293 (69.3%) of the participants had a high level of knowledge about the COVID- 19 vaccine (median knowledge score 6; IQR = 3), and 239 (56.5%) were reported to have a low level of vaccine acceptance (median acceptance scores 4; IQR=2). The knowledge level towards the COVID-19 vaccine was significantly associated with the vaccine acceptance level (p<0.001).
CONCLUSION: The community's level of knowledge towards COVID-19 vaccine was high; however, the vaccine acceptance was low.
Characteristics and performance of aerobic granular sludge treating rubber wastewater at different hydraulic retention time

Rosman NH, Nor Anuar A, Chelliapan S, Md Din MF, Ujang Z

Bioresour Technol, 2014 Jun;161:155-61.
PMID: 24704837 DOI: 10.1016/j.biortech.2014.03.047

The influence of hydraulic retention time (HRT, 24, 12, and 6h) on the physical characteristics of granules and performance of a sequencing batch reactor (SBR) treating rubber wastewater was investigated. Results showed larger granular sludge formation at HRT of 6h with a mean size of 2.0±0.1mm, sludge volume index of 20.1mLg(-1), settling velocity of 61mh(-1), density of 78.2gL(-1) and integrity coefficient of 9.54. Scanning electron microscope analyses revealed different morphology of microorganisms and structural features of granules when operated at various HRT. The results also demonstrated that up to 98.4% COD reduction was achieved when the reactor was operated at low HRT (6h). Around 92.7% and 89.5% removal efficiency was noted for ammonia and total nitrogen in the granular SBR system during the treatment of rubber wastewater.
Fulltext Allelopathic effects of litter Axonopus compressus against two weedy species and its persistence in soil

Samedani B, Juraimi AS, Rafii MY, Anuar AR, Sheikh Awadz SA, Anwar MP

ScientificWorldJournal, 2013;2013:695404.
PMID: 24260020 DOI: 10.1155/2013/695404

This study investigated the allelopathic effect of Axonopus compressus litter on Asystasia gangetica and Pennisetum polystachion. In experiment 1 the bioassays with 0, 10, 30, and 50 g L⁻¹ of aqueous A. compressus litter leachate were conducted. Experiment 2 was carried out by incorporating 0, 10, 20, 30, 40, and 50 g L⁻¹ of A. compressus litter leachate into soil. In experiment 3, the fate of A. compressus litter leachate phenolics in the soil was investigated. A. compressus leachates did not affect the germination percentage of A. gangetica and P. polystachion, but delayed germination of A. gangetica seeds and decreased seed germination time of P. polystachion. A. compressus litter leachates affected weeds hypocotyl length. Hypocotyl length reductions of 18 and 31% were observed at the highest concentration (50 g L⁻¹) compared to the control in A. gangetica and P. polystachion, respectively. When concentration of A. compressus litter leachate-amended soil increased A. gangetica and P. polystachion seedling shoot length, root length, seedling weight and chlorophyll concentration were not affected. The 5-week decomposition study of A. compressus showed that the phenolic compounds in A. compressus litter abruptly decreased about 52% after two weeks and remained steady until the end of the incubation.
Fulltext Competitive interaction of Axonopus compressus and Asystasia gangetica under contrasting sunlight intensity

Samedani B, Juraimi AS, Anwar MP, Rafii MY, Sheikh Awadz SH, Anuar AR

ScientificWorldJournal, 2013;2013:308646.
PMID: 24163618 DOI: 10.1155/2013/308646

Axonopus compressus is one of the native soft grass species in oil palm in Malaysia which can be used as a cover crop. The competitive ability of A. compressus to overcome A. gangetica was studied using multiple-density, multiple-proportion replacements series under a glasshouse and full sunlight conditions in a poly bag for 10 weeks. A. compressus produced more dry weight and leaf area when competing against A. gangetica than in monoculture at both densities in the full sunlight and at high density in the shade. Moreover, the relative yield and relative crowding coefficients also indicated A. compressus is a stronger competitor than A. gangetica at both densities in the full sunlight and high density in the shade. It seemed that A. gangetica plants in the shade did not compete with each other and were more competitive against A. compressus as could influence A. compressus height in the shade. It is concluded that although suppression of A. gangetica by A. compressus occurred under full sunlight, irrespective of plant density, this ability reduced under shade as A. compressus density decreased. The result suggests that A. compressus in high density could be considered as a candidate for cover crops under oil palm canopy.
Fulltext Marital status and its relationship with the risk and pattern of visual impairment in a multi-ethnic Asian population

Zheng Y, Lamoureux EL, Chiang PP, Rahman Anuar A, Wong TY

J Public Health (Oxf), 2014 Mar;36(1):104-10.
PMID: 23658389 DOI: 10.1093/pubmed/fdt044

To examine whether marital status is a significant determinant of visual impairment (VI) in urban multi-ethnic Asian population.
Sourcing different neuro-progenitor cell for the use of nerve construct

Yazid AG, Anuar A, Onhmar HT, Ng AM, Ruszymah BH, Amaramalar SN

Med J Malaysia, 2008 Jul;63 Suppl A:113-4.
PMID: 19025011

Spinal cord, sciatic nerve, olfactory ensheathing cell and bone marrow derived mesenchymal stem cells were evaluated as an alternative source for tissue engineering of nerve conduit. All cell sources were cultured in alpha-MEM medium. Olfactory Ensheathing Cell (OEC) showed the best result with higher growth kinetic compared to the others. Spinal cord and sciatic nerve were positive for GFAP, OEC were positive for GFAP, S100b and anti-cytokeratin 18 but negative for anti-Human Fibroblast.
A recombinant antigen-based IgG4 ELISA for the specific and sensitive detection of Brugia malayi infection

Rahmah N, Lim BH, Khairul Anuar A, Shenoy RK, Kumaraswami V, Lokman Hakim S, et al.

Trans R Soc Trop Med Hyg, 2001 8 9;95(3):280-4.
PMID: 11490997

An IgG4 ELISA based on a novel recombinant antigen was evaluated for detection of Brugia malayi infection, using 2487 sera from various institutions: 2031 samples from Universiti Sains Malaysia, 276 blinded sera from 2 other institutions in Malaysia, 140 blinded sera from India and 40 blinded sera from Thailand. These sera were from various groups of individuals, i.e., microfilaraemics, chronic patients, endemic normals, non-endemic normals and individuals with other parasitic and bacterial infections. Based on a cut-off optical density reading of 0.300, the IgG4 ELISA demonstrated specificity rates of 95.6-100%, sensitivity rates of 96-100%, positive predictive values of 75-100% and negative predictive values of 98.9-100%. These evaluation studies demonstrated the high specificity and sensitivity of this test for the detection of active B. malayi infection. Thus, the IgG4 ELISA would be very useful as a tool in diagnosis and in elimination programmes for brugian filariasis.
Fulltext Toxoplasmosis in HIV/AIDS patients in Malaysia

Nissapatorn V, Lee CK, Cho SM, Rohela M, Anuar AK, Quek KF, et al.

Southeast Asian J. Trop. Med. Public Health, 2003;34 Suppl 2:80-5.
PMID: 19238664

Three hundred and one sera of HIV/AIDS patients were tested for anti-Toxoplasma IgG antibody by ELISA technique. The seroprevalence of toxoplasmosis was 41.2% (95% CI: 35.5-46.9) in HIV/AIDS patients. The seroprevalence was significantly higher in the Malay (57.9%) than the Chinese (38.7%), followed by the Indian patients (29.6%) (p<0.05). No possible risk factor, such as contact with cats, consumption of uncooked meat, and history of blood transfusions was found to have any significant association with the presence of anti-Toxoplasma antibody in the study sample (p>0.05). Multivariate analysis was employed to find any association between Toxoplasma seroprevalence and a single subject having single or multiple risk factors. It was found that the association was not statistically significant (p>0.05). Among the HIV/AIDS study samples, 124 (41.2%) samples were found to have positive anti-Toxoplasma antibody, the association between the presence of anti-Toxoplasma antibody and CD4 cell count was determined but no statistically significant association was found (p>0.05). During the study period, only one case of active CNS toxoplasmosis was registered and the diagnostic criteria included: clinical presentations, CT scan finding, serological evidence of anti-Toxoplasma IgG antibody, and respose to anti-Toxoplasma therapy.
Fulltext Seroepidemiology of toxoplasmosis among HIV-infected patients and healthy blood donors

Nissapatorn V, Kamarulzaman A, Init I, Tan LH, Rohela M, Norliza A, et al.

Med J Malaysia, 2002 Sep;57(3):304-10.
PMID: 12440270 MyJurnal

A cross-sectional study was carried out in University of Malaya Medical Centre, Kuala Lumpur. Blood samples from 100 HIV-infected patients and 203 Healthy Blood Donors (HBD) were collected and anti-Toxoplasma antibodies were detected by using conventional ELISA. The seroprevalence of toxoplasmosis in HIV/AIDS and Healthy Blood Donors were found to be 21% and 28.1% respectively. There was no significant association between the seroprevalence of toxoplasmosis and various possible risk factors i.e. contact with cat, consumption of undercooked meat and history of blood transfusion in both groups. No significant differences between Toxoplasma seroprevalence in HIV/AIDS and Healthy Blood Donors in association with presence of single or multiple risk factors were found. The mean CD4 count among HIV/AIDS patients in this study was 202.23 cell/cumm. There was no significant association between CD4 count and seropositivity for Toxoplasma antibodies in HIV/AIDS patients.
Blastocystis in animal handlers

Rajah Salim H, Suresh Kumar G, Vellayan S, Mak JW, Khairul Anuar A, Init I, et al.

Parasitol Res, 1999 Dec;85(12):1032-3.
PMID: 10599928

The present study investigated whether people working closely with animals were at higher risk of getting infected with Blastocystis hominis. The prevalence of the parasite was determined in two population groups, i.e., animal handlers and normal healthy individuals who did not work with animals. In all, 105 stool samples were collected from animal handlers from 2 local research institutions, a local zoo, and a local abattoir and 163 stool samples were collected from normal healthy individuals residing in high-rise flats in the city. The in vitro culture method used in the study detected that 41% of 105 animal handlers and 17% of 163 flat-dwellers in the city were positive for Blastocystis. This statistically significant finding (P = 0.0000313) shows that people who work closely with animals do stand at risk of acquiring Blastocystis infection.
PCR-ELISA for the detection of Brugia malayi infection using finger-prick blood

Rahmah N, Ashikin AN, Anuar AK, Ariff RH, Abdullah B, Chan GT, et al.

Trans R Soc Trop Med Hyg, 1998 12 16;92(4):404-6.
PMID: 9850392

A polymerase chain reaction assay based on the enzyme-linked immunosorbent assay (PCR-ELISA) has been developed to detect Brugia malayi infection in an area of low endemicity in Malaysia. Blood samples from 239 subjects were tested: 192 amicrofilaraemic individuals, 14 microfilaraemic persons and 3 chronic elephantiasis cases from endemic areas and 30 city-dwellers (non-endemic controls). PCR products were examined by ELISA and Southern hybridization. In the PCR-ELISA, digoxigenin-labelled PCR products were hybridized to a biotin-labelled probe. This was followed by incubation in streptavidin-coated microtitre wells and detection using anti-digoxigenin-peroxidase and ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulphonic acid)]. All microfilaraemic samples were positive by PCR-ELISA and Southern hybridization and all samples from non-endemic subjects and chronic elephantiasis patients were negative. The PCR-ELISA detected 12 times as many B. malayi infections as did thick blood film examination. Nineteen of the 194 samples from the endemic area gave positive results by both PCR-ELISA and Southern hybridization, and an additional 5 samples were positive by PCR-ELISA only. The PCR-ELISA was specific and sensitive, detected more infections, and was more reproducible than Southern hybridization.
Helicobacter pylori infection in north-eastern peninsular Malaysia. Evidence for an unusually low prevalence

Uyub AM, Raj SM, Visvanathan R, Nazim M, Aiyar S, Anuar AK, et al.

Scand. J. Gastroenterol., 1994 Mar;29(3):209-13.
PMID: 8209178

The prevalence of Helicobacter pylori infection was determined in peptic ulcer patients, in non-ulcer dyspepsia (NUD) patients, and in the general adult population. The H. pylori infection rate ascertained by microbiologic examination of multiple gastric antral biopsy specimens was 50% (17 of 34) in duodenal ulcer (DU), 5% (1 of 22) in gastric ulcer, and 9% (15 of 159) in NUD patients. A seroepidemiologic survey showed a prevalence of only 4.2% among 496 blood donors and 4.8% among 921 subjects who attended health screening clinics. H. pylori infection is relatively uncommon and does not appear to be the predominant factor in the pathogenesis of peptic ulcer disease in the area. The incidence of peptic ulcer perforations in the area in 1991-92 was 1.5 per 100,000 person-years, reflecting a relatively low frequency of peptic ulcers, which might be due to the low prevalence of H. pylori infection in the population.
A Brugia malayi antigen specifically recognized by infected individuals

Rahmah N, Anuar AK, A'shikin AN, Lim BH, Mehdi R, Abdullah B, et al.

Biochem Biophys Res Commun, 1998 Sep 29;250(3):586-8.
PMID: 9784388

Western blot analyses were performed on 444 serum specimens: 40 sera from microfilaraemic individuals, 10 sera from elephantiasis patients, 24 treated individuals, 50 sera from residents of endemic areas without anti-filarial IgG4 antibodies (endemic normals), 20 sera from amicrofilaraemic individuals with high anti-filarial IgG4 antibodies, 200 sera from healthy city-dwellers (non-endemic samples), and 100 sera from soil-transmitted helminth-infected individuals. Phast electrophoresis system was used to electrophorese Brugia malayi soluble adult worm antigen on 10-15% SDS-PAGE gradient gels followed by electrophoretic transfer onto PVDF membranes. Membrane strips were then successively incubated with blocking solution, human sera, and monoclonal anti-human IgG4 antibody-HRP, with adequate washings done in between each incubation step. Luminol chemiluminescence detection was then used to develop the blots. An antigenic band with the MW of approximately 37 kDa was found to be consistently present in the Western blots of all microfilaraemic sera, all amicrofilaraemic sera with high titres of anti-filarial IgG4 antibodies, some treated patients, and some elephantiasis patients. The antigen did not occur in immunoblots of individuals with other helminthic infections, normal endemic individuals, and city dwellers. Therefore the B. malayi antigen of with the MW of approximately 37 kDa demonstrated specific reactions with sera of B. malayi-infected individuals and thus may be useful for diagnostic application.
Polypeptides associated with in vitro cyst formation of Blastocystis hominis

Init I, Mak JW, Top S, Zulhainan Z, Prummongkol S, Nissapatorn V, et al.

Southeast Asian J. Trop. Med. Public Health, 2003 Dec;34(4):727-32.
PMID: 15115079

The objective of this study was to characterize the polypeptides associated with cysts of Blastocystis hominis. This form is believed to be infective and plays a role in parasite resistance to anti-B. hominis drugs currently used for treatment of Blastocystis associated diarrhea. Cysts were induced through in vitro culture of the parasite in complete medium supplemented with bacterial extract with trypticase, metronidazole or doxycycline. SDS-PAGE analysis showed almost similar polypeptide patterns of parasite extracts obtained from in vitro cultured parasites before and after exposure with the three supplements. Polypeptide bands at 76, 58.5, 48, 45, 40, 38, 32, 25 and 22 kDa were constantly seen in all antigenic preparations and no specific cyst-associated polypeptide was present. However, on immunoblot analysis, 3 out of 16 blastocystosis human sera identified a cyst-associated polypeptide at 60 kDa in all parasite extracts prepared from cultures with the three supplements. In addition, there were associated morphological changes detected in these parasites stained with acridine orange and observed under fluorescence microscopy. Metronidazole induced cyst forms (reddish cells) as early as 12 hours post-exposure; more cyst production (with stronger immunoblot bands) occurred after 24 hours exposure. However, cysts rupture with release and destruction of B. hominis daughters cells occurred after 48 hours exposure. Doxycycline induced less cyst-like forms at 24 hours (weaker 60 kDa band) and less destruction of the cysts (60 kDa band still present at 72 hours post exposure). Bacterial extract and trypticase also induced cysts at 12 hours with increasing numbers up to 72 hours exposure (corresponding increase in intensity of 60 kDa band from samples harvested at 12 to 72 hours post exposure) without any sign of deleterious effect on the parasite.

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