Agar Plate Culture Method Research Articles

Bacterial Community of Breast Milk in Breastfeeding Women Using Culture-Dependent and Culture-Independent Approaches.

This study aimed to analyze bacterial communities in breast milk obtained from five breastfeeding women. Culture-dependent and culture-independent methods were used to analyze microbial communities. Total bacterial count of breast milk determined using plate count agar ranged from 3.3 × 104 ± 3.5 × 102 colony forming unit (CFU)/g to 1.7 × 105 ± 3.5 × 103 CFU/g, with a pH between 6.4 and 6.8. Only three species, Leuconostoc citreum (17 out of 160 strains; 10.63%), Staphylococcus epidermidis (118 strains; 73.75%), and Staphylococcus lugdunensis (25 strains; 15.63%), belong to the phylum Bacillota were detected by culture-dependent analysis. Microbial communities analyzed via pyrosequencing revealed greater diversity compared to the culture-dependent analysis. At the phylum level, Bacillota accounted for 60.9% of the microbial community. At the genus level, Staphylococcus (24.57%), Streptococcus (22.93%), and Methylobacterium (8.76%) were dominant genera. While pyrosequencing demonstrated greater microbial diversity than the agar plate culture method, identified microbes might lack information or include many unculturable microbes. Most of all, considering the low total bacterial count averaging 7.2 × 104 CFU/g, further research is needed to determine the significance of microbial presence in breast milk.

Comparison of parasitological methods for the identification of soil-transmitted helminths, including Strongyloides stercoralis, in a regional reference laboratory in northwestern Argentina: An observational study

Soil-transmitted helminths (STH) are a significant public health problem in impoverished communities of tropical and subtropical areas. Improved diagnostic methods are crucial for Neglected Tropical Diseases programs, particularly for S. stercoralis, as traditional methods are inadequate. Thus, it is important to identify the most accurate and efficient methods for the diagnosis of STH. We performed a retrospective study analyzing laboratory data at the Instituto de Investigaciones de Enfermedades Tropicales from 2010 to 2019. The study included data from outpatients referred for stool analysis and public health interventions from urban and rural communities in northern Salta province, Argentina. Samples were included in this analysis if processed through sedimentation/concentration, Baermann, Harada-Mori and McMaster's, with a subgroup that also included Agar plate culture method (APC). Sensitivity was calculated against a composite reference standard. Of the 5625 samples collected, 944 qualified for this analysis, with a prevalence of 11.14% for A. lumbricoides, 8.16% for hookworm, 1.38% for T. trichiura, and 6.36% for S. stercoralis. The sedimentation/concentration method was the most sensitive for A. lumbricoides (96%), compared to the McMaster method, with a sensitivity of 62%. Similarly, for hookworms, sedimentation/concentration was more sensitive than McMaster's, Harada-Mori, and Baermann with sensitivities of 87%, 70%, 43%, and 13%, respectively. Most of these infections were of light intensity. For S. stercoralis, Baermann and sedimentation/concentration methods were the most sensitive, with 70% and 62% respectively, while Harada-Mori was the least sensitive. In a subset of 389 samples also analyzed by the APC, Baermann was more sensitive than APC for detecting S. stercoralis, and both methods were superior to Harada-Mori. Parasitological methods, mostly when used combined, offer adequate opportunities for the diagnosis of STH in clinical and public health laboratories. The incorporation of S. stercoralis into the control strategies of the World Health Organization requires rethinking the current diagnostic approach used for surveys. With sedimentation/concentration and Baermann appearing as the most sensitive methods for this species. Further studies, including implementation assessments, should help in identifying the most adequate and feasible all-STH diagnostic approach.

The enhancive effect of the 2014-2016 El Niño-induced drought on the control of soil-transmitted helminthiases without anthelmintics: A longitudinal study.

Soil-transmitted helminthiases (STHs) are common in tropical and subtropical regions. Southern Thailand experiences an extended rainy season, leading to persistently moist soil. This condition supports the life cycle of STHs, hindering effective control due to reinfection and low drug efficacy. We implemented a novel STH control strategy during the dry season aimed at decreasing reinfection rates without enhancing sanitation or hygiene practices. However, there were unexpected, prolonged droughts linked to El Niño events from 2014 to 2016. Additionally, we assessed the effects of these drought conditions on further control measures without the use of anthelmintics. A longitudinal study was conducted from 2012 to 2016. Stool samples collected from 299 participants were analyzed using the Kato-Katz and agar plate culture methods. Participants who tested positive for STHs received a single 400 mg dose of albendazole. The efficacy of the treatment was evaluated three weeks later. To confirm the control measures were implemented during the dry season, we monitored the number of rainy days following albendazole treatment for 52 days, of which 38 were without rain. Follow-up stool examinations were carried out in 2013 and 2016, with no additional doses of albendazole administered. Rainfall and rainy day data, which served as indicators of unexpected droughts due to El Niño, were collected from the nearest local meteorological stations. Before the drought, there was a decrease in STH prevalence in 2013-except for trichuriasis-attributable to the dry season control efforts. Despite these efforts, STH prevalence remained high. Remarkably, in 2016, following the drought period, the prevalence of trichuriasis, which had not changed previously, spontaneously declined without further albendazole treatment compared to 2013. Furthermore, the prevalence of strongyloidiasis remained unchanged likely due to its low susceptibility to drought conditions, as it can reproduce within hosts. Conversely, the prevalence of other STHs consistently declined. The drought and possible improvements in sanitation and hygiene practices contributed to this decrease by reducing rates of reinfection and new infection and by increasing the natural cure rate. Additionally, some participants infected with hookworms or Trichuris who were not cured by albendazole experienced natural remission. Control measures implemented during the dry season, combined with a 14-month-long drought induced by the El Niño event of 2014-2016, and some improvements in sanitation and hygiene practices, contributed to a decrease in both the prevalence and intensity of STHs, except for S. stercoralis. Over time, S. stercoralis is likely to become the predominant species among the STHs.

Evaluation of five diagnostic methods for Strongyloides stercoralis infection in Amhara National Regional State, northwest Ethiopia

BackgroundStrongyloides stercoralis is an intestinal parasite that can cause chronic infection, hyperinfection and/or a dissemination syndrome in humans. The use of techniques targeting ova fails to detect S. stercoralis, as only larvae of the parasite are excreted in faeces. Due to the absence of “Gold” standard diagnostic method for S. stercoralis, there is a paucity of reported data worldwide.ObjectiveThis study aimed to evaluate the performance of diagnostic methods of S. stercoralis infection by taking the composite reference as a “Gold” standard.MethodsA cross-sectional study was conducted among 844 schoolchildren in Amhara Region, Ethiopia, from April to December 2019. Stool samples were collected and processed with formol-ether concentration technique (FECT), spontaneous tube sedimentation technique (STST), Baermann concentration technique (BCT), agar plate culture (APC) and real-time polymerase chain reaction (RT-PCR). Sensitivity, specificity, positive predictive value, and negative predictive value of each diagnostic method were computed against the composite reference. The agreements of diagnostic methods were evaluated by Kappa value at 95% CI.ResultsThe composite detection rate of S. stercoralis by the five diagnostic methods was 39.0% (329/844). The detection rate of the parasite from stool samples by FECT, STST, BCT, APC and RT-PCR was 2.0% (17/844), 4.0% (34/844), 10.2% (86/844), 10.9% (92/844) and 28.8% (243/844), respectively. The highest detection rate (37.8%; 319/844) of S. stercoralis was recorded by a combination of BCT, APC, and RT-PCR followed by a combination of STST, BCT, APC and RT-PCR (37.3%; 315/844). The sensitivity of FECT, STST, BCT, APC and RT-PCR against the composite reference was 5.2%, 10.3%, 26.4%, 28.0% and 73.9%, respectively. The diagnostic agreements of RT-PCR, APC, BCT, STST and FECT with the composite reference in detection of S. stercoralis were substantial (0.775), fair (0.321), fair (0.305), slight (0.123), and slight (0.062), respectively.ConclusionRT-PCR detected the highest number of S. stercoralis infections. A combination of RT-PCR with APC and/or BCT better detected S. stercoralis from stool samples compared to other combinations or single diagnostic methods. Therefore, RT-PCR and combination of RT-PCR with APC and/or BCT diagnostic methods should be advocated for detection of S. stercoralis infection.

Development of a device for cultivation and isolation of microbes using a specialized cellulose film

A device was developed for the cultivation and isolation of microbes using a specialized cellulose film (SCF). This method can solve problems associated with the agar plate method. It can also be used for other culture-related operations, which is not possible with the traditional agar plate culture method.

Importance of detection of Strongyloides stercoralis DNA in fecal samples from patients with type 2 diabetes mellitus

ObjectiveThe association between diabetes and Strongyloides infection remains controversial. This study aimed to detect Strongyloides stercoralis DNA in the feces of patients with Diabetes Mellitus type 2 (DM2). MethodsFecal samples were analyzed via the Lutz, Rugai, and agar plate culture methods. PCR amplification was performed using two targets (PCR-genus and PCR-species) located on the S. stercoralis 18S ribosomal. ResultsThe positivity for S. stercoralis using parasitological methods was 1.1%. PCR-genus (14.13%) demonstrated a higher positivity than PCR-species (9.78%). ConclusionThe results confirm the greater positivity of the molecular diagnosis in relation to parasitological methods, reinforcing its use as an additional tool for the diagnosis of S. stercoralis infection in patients with DM2 living in endemic areas for this helminthiasis.

Evaluating the performance of diagnostic methods for soil transmitted helminths in the Amhara National Regional State, Northwest Ethiopia

BackgroundSoil-transmitted helminths are more prevalent in tropics and sub-tropics including Ethiopia. Despite their high prevalence, direct saline microscopy with its low sensitivity has been used as a diagnostic method in almost all health facilities in Ethiopia. Alternative diagnostic methods which have higher sensitivity are not yet implemented. Therefore, this study aimed to compare and evaluate the performance of diagnostic methods for soil transmitted helminths.MethodsA cross-sectional study among 520 school children was conducted from October to December, 2019 in Amhara National Regional State. The study participants were selected using systematic random sampling technique. Stool samples were processed via formol ether concentration, Kato-Katz, spontaneous tube sedimentation and agar plate culture techniques. Data was entered into Epi-data version 3.1 and analysis was done using SPSS version 20.0. The sensitivity, specificity, and negative predictive value were calculated against the combined result. Strength of agreement of the diagnostic methods was determined by Kappa value.ResultsThe Overall prevalence of soil transmitted helminths was 40.8% using combination of methods. The prevalence 24.4, 22.5, and 32.4%, respectively was recorded by using formol ether concentration, Kato-Katz and spontaneous tube sedimentation. The highest prevalence of hookworm (29.2%) was detected by the agar plate culture. The sensitivity and negative predictive value of formol ether concentration were 57.9 and 78.4%, for Kato-Katz thick smear 55.2 and 76.4%, for spontaneous tube sedimentation were 79.2 and 87.5% to soil transmitted helminths detection, respectively. The sensitivity and negative predictive value of agar plate culture to hookworm detection were 86.4 and 93.5%, respectively.ConclusionSpontaneous tube sedimentation shows higher sensitivity in the detection of soil transmitted helminth infections. Agar plate culture method also indicated better performance for hookworm detection than other methods. Therefore, the employment of spontaneous tube sedimentation technique for routine laboratory and agar plate culture for research purposes will significantly aid in accurate diagnosis of parasitic infections.

Determination of available breaking stress of agar and gellan gum plate culture methods and the duration of bacterial culture under strong acidic conditions.

Several acidophilic bacteria have not been cultured, primarily owing to the lack of suitable culture methods under strong acidic conditions. This study aimed to quantitatively evaluate the strengths of the agar plates (AP) and gellan gum plates (GP), and optimal culture periods under strong acidic conditions. To define the lower limit of plate strength for bacterial isolation culture, the diameter of Escherichia coli K12 colonies and the breaking stress of plates at different concentrations of gelling agents, medium composition and pH conditions were determined. The lower limit of available strength of AP and GP was 19·6 and 14·8kPa, respectively. Medium composition slightly affected AP breaking stress, although GP with a high cationic concentration medium could not be prepared. Assessment of the strength limits of AP and GP revealed that AP is not suitable for prolonged bacterial culture (≥72h). Furthermore, GP was completely ineffective for bacterial culture under highly acidic conditions (≤pH 1·0). Our quantitative evaluation method based on breaking stress is a potentially valuable tool to understand the state and the suitable limit of plate culture methods in more detail under various conditions.

Strongyloides stercoralis in Alcoholic Patients: Implications of Alcohol Intake in the Frequency of Infection and Parasite Load.

Strongyloides stercoralis infection in immunocompromised subjects, including chronic alcoholics, can lead to a severe disease. Moreover, its prevalence in alcoholic patients seems to be higher than that in the general population. The aims of this study were to evaluate the frequency of S. stercoralis infection in alcoholic patients and to investigate the influence of alcohol intake on the parasite load, as well as to evaluate the sensitivity of three different parasitological methods according to the larval output. Fecal samples of 1290 chronic alcoholic patients were examined by spontaneous sedimentation, Baermann–Moraes, and agar plate culture (APC) methods. S. stercoralis was the most frequent parasite found (14.5%; n = 187). Alcoholic individuals infected with Strongyloides stercoralis had a higher daily consumption of alcohol than those who were not infected, 528.6 and 403.0 g/day, respectively (p < 0.05). In addition, individuals with higher alcohol intake presented an increase in parasite load. The S. stercoralis diagnostic method with the highest sensitivity was APC, 97.9% (183/187). In conclusion, S. stercoralis seems to be the most frequent parasite found in alcoholic individuals from endemic areas and alcohol intake is positively associated with S. stercoralis larvae output. In addition, this study confirms that APC is the most sensitive parasitological method used for Strongyloides diagnosis.

Investigating the prevalence of intestinal parasites with an emphasis on Strongyloides stercoralis infection in hospitalized patients: a regional report from Iran.

Intestinal parasitic infections such as strongyloidosis are more common among individuals with immune deficiency and sometimes accompanied by severe symptoms. The purpose of this cross-sectional study was to evaluate the prevalence of intestinal parasites with focus on strongyloidosis in hospitalized patients. A total number of 566 faecal samples were obtained from different wards and assessed by the use of direct smear, formalin-ether concentration, and agar plate culture procedures in order to find parasitic protozoa and helminthes. The findings revealed that 10.1% (n=57) of the examined samples were positive for intestinal parasites. The highest prevalence rate was related to Entamoeba coli (4.6%, n=26) and the lowest one was related to Strongyloides stercoralis (0.5%, n=3). In addition, Giardia lamblia prevalence rate was 3.2% (n=18) and the prevalence rate of Blastocystis hominis was 1.8% (n=10). The sensitivity of S. stercoralis diagnosis was equal for agar plate culture and formalin-ether concentration methods. This study demonstrated the significance of focus on intestinal parasites in hospitalized patients and highlighted the necessity of improving the insight in health care providers about the occurrence of parasitic infections especially strongyloidiasis in these patients.

Prevalence and associated risk factors of Strongyloides stercoralis infection in Lower Myanmar

BackgroundStrongyloidiasis is prevalent in Southeast Asian regions along with other soil-transmitted helminthiases, but only limited present-day data was available for Myanmar.MethodsA prevalence survey for Strongyloides stercoralis infection was conducted among villagers in rural areas of three townships located in the Lower Myanmar during 2014–2016 by agar plate culture method in combination with specific identification by molecular assays. Risk factors associated with S. stercoralis infection were assessed by analyzing questionnaires obtained from study participants.ResultsStrongyloides stercoralis was identified in 40 out of 703 participants (5.7% overall prevalence). The highest prevalence (14.4%) was observed in Htantabin, while other two communities (Thabaung and Thanlyin) had much lower prevalence (2.2 and 2.5%, respectively). Infection was relatively rare (1.2%) in younger generations under 20 years compared to older generations (9.5%). Even in Htantabin, none of the female residents under age 40 (n = 33) had infection. In adult Htantabin residents, those who answered that they do not wear shoes regularly had an elevated risk of infection (odds ratio = 2.50, 95% confidence interval = 1.03–6.08).ConclusionsThis study showed that there is still an on-going transmission of strongyloidiasis in Lower Myanmar. It is highly desirable that the soil should be free of fecal contamination by improving the management of fecal waste. Meanwhile, health education to promote shoe-wearing would be beneficial to reduce the risk of transmission, especially for those who have frequent and intense contact with soil.

Is the Baermann, an applicable method for detection of Strongyloides stercoralis?

Introduction: It is estimated that 100 - 300 million people in the world are infected with Strongyloides stercoralis. Although there are several laboratory diagnostic methods for detection of this parasite, there is still no gold standard diagnostic method. The need for an application test with high sensitivity and specificity is highly felt, especially in people with immune deficiency or organ transplant volunteers. Methods: The diagnostic sensitivity of five parasitological methods, including direct smear of feces in saline and Lugol iodine stain, Baermann technique, formalin-ethyl acetate concentration, Harada-Mori filter paper, agar plate culture and molecular method was evaluated by stool specimens collected from the central mentally retardation institute of Bandar Abbas, to find out an easy, practical and cost-effective method for detecting Strongyloides stercoralis. Results: Five parasitological and molecular methods were investigated using 163 stool samples. Considering the employed methods, there found to be 30 cases of S. stercoralis. The total positive cases of S. stercoralis using different methods was considered as golden standard. The highest sensitivity of parasitological methods was associated with the Baermann 27 (90) and the least positive results obtained with Harada-Mori method, 1 (3.3). In this study 9 positive cases were detected by PCR method (30). Conclusion: Highest sensitivity was observed when different diagnostic approaches were combined where Baermann technique proceeded it. Therefore, this technique is recommended as a selective method for detecting S. stercoralis due to its easy, practical and inexpensive nature. n

Comparative evaluation of harada–Mori and agar plate culture for the identification of hookworm species under limited resources

Background: Human hookworm infection has widespread socioeconomic and public health implications. Several coproculture techniques have been developed for morphological identification of hookworm larvae under limited resource availability. The objective of this study was to compare the performances of Harada–Mori culture (HMC), agar plate culture (APC), and modified APC (MAPC) of hookworm positive stool specimen for identification of hookworm species occurring in East Sikkim, India. Materials and Methods: This prospective cohort study was performed using 180 stool specimen collected from children who attended Central Referral Hospital and Sir Thodup Namgyal Memorial Hospital, with the complained of gastrointestinal symptoms. Blood samples were also collected to correlate with the complete blood count (CBC). The hookworm positive stool specimen evaluated by microscopy was subjected to HMC, APC, and MAPC techniques to harvest hookworm larvae. Stoll's dilution egg count for determining egg intensity and CBC were also performed for the children who were positive for hookworm's eggs in their stool sample. Results: This study observed a predominance of Necator americanus (75%) over Ancylostoma duodenale (25%). CBC results showed high packed cell volume values in 9/12, low hemoglobin 9/12, and high eosinophil count in all the positive children. Stoll's dilution egg count showed moderate infection in 66.6%, light and heavy infections in 16.7% of children's. APC method was superior to HMC and MAPC in culturing and identifying hookworm species. Conclusions: APC was observed to yield better results and was easier to perform in limited resource laboratory setting compare to MAPC or Harada–Mori culture techniques.

First molecular identification and genetic diversity of Strongyloides stercoralis and Strongyloides fuelleborni in human communities having contact with long-tailed macaques in Thailand.

The parasitic nematodes, Strongyloides stercoralis and Strongyloides fuelleborni, can infect humans and non-human primates. We amplified and sequenced a portion of the 18S ribosomal RNA gene (rRNA) and of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene of Strongyloides from humans in the study area in Thailand, where people have frequent contact with long-tailed macaques. Fresh stool samples were obtained from 213 people and were examined using the agar plate culture method. The overall prevalence of Strongyloides infection was 8.92% (19/213). From a total of 19 worms (one per infected person), 18 adult males had 18S rRNA sequences identical with that of S. stercoralis and one adult female had a sequence almost identical with that of S. fuelleborni. A median-joining network of cox1 sequences revealed nine new haplotypes from S. stercoralis, and an overall haplotype diversity (Hd) of 0.9309. The single haplotype of S. fuelleborni was also new and contributed to an overall haplotype diversity for that species of 0.9842. This is the first molecular identification of S. stercoralis and S. fuelleborni in a human community having contact with long-tailed macaques in Thailand. It is also the first report of S. fuelleborni infecting a human in Thailand.

Porphyromonas gingivalis can invade periodontal ligament stem cells

BackgroundPorphyromonas gingivalis is strongly associated with the development, progression, severity and recurrence of periodontitis. Periodontal ligament stem cells (PDLSCs) play an important role in the maintenance of periodontal tissue self-renewal and repair. The purpose of this study was to investigate the ability of P. gingivalis to infect PDLSCs using an in vitro monolayer model.MethodsWe separated and cultured primary PDLSCs using the tissue block with limiting dilution method. The efficiency of P. gingivalis (ATCC 33277) infection of PDLSCs was measured using agar plate culture and quantitative polymerase chain reaction (q-PCR) methods. PDLSCs infected with P. gingivalis were also observed by transmission electron microscopy.ResultsWe assessed stem cell properties including cell morphology, clone formation, growth activity, cell surface antigens and multiple differentiation capacity. The infection rates of P. gingivalis in PDLSC at MOIs of 50, 100, 200, and 500 were 5.83%, 8.12%, 7.77% and 7.53% according to the agar plate culture method. By q-PCR, the efficiencies of P. gingivalis infection of PDLSCs at MOIs of 50, 100, 200, and 500 were 6.74%, 10.56%, 10.36% and 9.78%, respectively. Overall, the infection efficiency based on q-PCR was higher than that according to agar plate culture. Using transmission electron microscopy, we verified that P. gingivalis (ATCC 33277) could infect and invade PDLSCs after 2 h of incubation, and endocytic vacuoles were not found surrounding the internalized bacteria.ConclusionsIn conclusion, our data demonstrate that P. gingivalis can invade PDLSCs.

The roundworm Strongyloides stercoralis in children, dogs, and soil inside and outside a segregated settlement in Eastern Slovakia: frequent but hardly detectable parasite.

A comparative study was carried out to evaluate the Strongyloides stercoralis infections in children and dogs inside and outside the segregated settlement in Medzev, Eastern Slovakia, and a survey of the soil within the settlement was included. Applying the Koga agar plate (KAP) culture method and microscopy examination of stool samples collected from 60 Roma and 21 nonRoma children, no larvae of S. stercoralis were detected but eggs of three nematodes (Ascaris lumbricoides, Trichuris trichiura, and Enterobius vermicularis) and cysts of two protozoan endoparasites (Giardia duodenalis and Cryptosporidium spp.) were often found. However, immunoenzymatic assay (ELISA) for the evidence of IgG antibodies against S. stercoralis showed 33.3% seroprevalence in Roma children and 23.8% prevalence in children from the majority population, attending the same school. Eosinophilia was regularly present in children with exclusive infection of S. stercoralis (eight cases) as well as in individuals suffering from mixed infections of S. stercoralis and some of the above listed parasites (16 cases); high eosinophil counts sometimes, but not always, occurred in parasitized children lacking S. stercoralis antibodies. A comparison of S. stercoralis in dogs from the settlement (40 dogs) and from a distant dog shelter (20 dogs) did not reveal remarkable differences: the direct microscopy of faecal samples revealed rhabditiform larvae in 13.3% of the dogs from the settlement (4/30) and in 10.0% of the dogs from the shelter (2/20). Out of blood samples collected from the second dog group, 55% of the dogs contained antibodies against S. stercoralis. In the soil collected from 14 various locations within the settlement, S. stercoralis larvae were observed in two samples (14.3%); however, 13 samples (92.9%) were positive for human or dog endoparasites of the genera Ancylostoma, Ascaris, Toxocara, Toxascaris, Trichuris, and Hymenolepis.

First molecular identification and report of genetic diversity of Strongyloides stercoralis, a current major soil-transmitted helminth in humans from Lao People's Democratic Republic.

Strongyloidiasis is a major soil-transmitted helminth (STH) disease that affects people worldwide. We present updated data on prevalence in the Lao People's Democratic Republic (Lao PDR) in 2015, arising from a community cross-sectional helminthiasis survey. Fecal samples were collected from 327 individuals across three provinces in Lao PDR (Luang Prabang in the north, Khammouane in the center, and Champasack in the south). Agar plate culture and Kato-Katz methods were used to examine duplicate stool samples from each participant to detect Strongyloides stercoralis and co-infecting helminths. Overall prevalences of S. strercoralis human hookworm, Taenia spp., Trichuris trichiura, Ascaris lumbricoides, and Enterobius vermicularis were 41.0, 28.1, 4.9, 4.0, 1.5, and 0.9%, respectively. The prevalence of miscellaneous trematodiases (including opisthorchiasis) was 37.9% and of Schistosoma mekongi infection was 0.3%. Strongyloidiasis is a current major STH disease in Lao PDR. We also report the molecular-phylogenetic identification of S. stercoralis adult males collected from 40 representative human strongyliodiasis fecal samples. DNA was extracted, amplified, and sequenced from a portion of the mitochondrial cox1 gene and the nuclear 18S ribosomal DNA. Phylogenetic analyses indicated that all specimens sequenced belonged to S. stercoralis (Bavay, 1876) Stiles and Hassall, 1902. The cox1 sequences exhibited great diversity (24 haplotypes) in Lao PDR. This is the first molecular identification and report of genetic diversity of S. stercoralis in humans from Lao PDR. An effective parasite control program is needed to reduce the serious health impacts.

DIAGNOSIS OF Strongyloides stercoralis INFECTION IN IMMUNOCOMPROMISED PATIENTS BY SEROLOGICAL AND MOLECULAR METHODS.

SUMMARYStrongyloidiasis is a potentially serious infection in immunocompromised patients. Thus, the availability of sensitive and specific diagnostic methods is desirable, especially in the context of immunosuppressed patients in whom the diagnosis and treatment of strongyloidiasis is of utmost importance. In this study, serological and molecular tools were used to diagnose Strongyloides stercoralis infections in immunosuppressed patients. Serum and stool samples were obtained from 52 patients. Stool samples were first analyzed by Lutz, Rugai, and Agar plate culture methods, and then by a quantitative real time polymerase chain reaction (qPCR). Serum samples were evaluated by an enzyme-linked immunosorbent assay (ELISA) using a soluble (AS) or a membrane fractions antigen (AM) obtained from alkaline solutions of the filariform larvae of Strongyloides venezuelensis. Of the 52 immunosuppressed patients, three (5.8%) were positive for S. stercoralis by parasitological methods, compared to two patients (3.8%) and one patient (1.9%) who were detected by ELISA using the AS and the AM antigens, respectively. S. stercoralis DNA was amplified in seven (13.5%) stool samples by qPCR. These results suggest the utility of qPCR as an alternative diagnostic tool for the diagnosis of S. stercoralis infection in immunocompromised patients, considering the possible severity of this helminthiasis in this group of patients.

Strongyloides stercoralis infection in patients with systemic lupus erythematosus: diagnosis and prevention of severe strongyloidiasis.

Strongyloides stercoralis infection is usually chronic and asymptomatic and may persist undiagnosed for decades. However, in immunocompromised individuals, the infection can cause hyperinfection and dissemination. Therefore, early diagnosis is essential to prevent severe forms of strongyloidiasis. The aims of this study were: (i) to evaluate the frequency of S.stercoralis infection in patients with systemic lupus erythematous (SLE) and (ii) to estimate specific immunoglobulins G (IgG) and E (IgE) production using an enzyme-linked immunosorbent assay (ELISA) method. Seventy-five SLE patients treated with prophylactic anthelmintic therapy were evaluated using the spontaneous sedimentation (SS), Baermann-Moraes (BM) and agar plate culture (APC) methods. Serum anti-S.stercoralis IgG and IgE antibodies were measured by ELISA. Using parasitological methods, the frequency of intestinal parasites was 10.7%, whereas the frequency of S.stercoralis infection was 1.3%. The sensitivity of the ELISA to detect anti-S.stercoralis IgG and IgE was 80% and 76.9%, respectively. Both assays presented the same specificity of 96.7%. The frequency of anti-S.stercoralis IgG and IgE was 16% and 28%, respectively. Six patients were positive for both antibodies. Diagnostic approaches using high-sensitivity parasitological methods and the detection of specific antibodies are essential for the diagnosis of strongyloidiasis in immunocompromised patients. Early detection of infection can alter the course of the disease via appropriate treatment, preventing the occurrence of severe strongyloidiasis.

Modified agar plate culture method for culture of Strongyloides stercoralis.

Modified agar plate culture method for culture of Strongyloides stercoralis.