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Background Giardia spp. is a protozoan pathogen and is the most common enteric parasite of domestic animals and humans. Assays for detecting infection in fecal samples using direct or indirect examinations are important tools for diagnosing the disease. The objective of the present study was to compare the cost-effectiveness of immunoassays and FLOTAC technique for diagnosing Giardia spp. infection in dogs. Results Fecal samples from 80 positive stray dogs were tested for the presence of copro-antigens of Giardia spp. using the direct immunofluorescence assay (IFA), a rapid enzyme-linked immunosorbent assay (ELISA) and the FLOTAC double technique. All methods were performed in accordance with the instructions reported in the original description for each technique. The results showed that ELISA can be run in less time than IFA and almost at the same time of the FLOTAC technique. Among the tests used in this study, FLOTAC had the lowest cost per correct diagnosis, compared with immunoassays. Conclusions The results from this cost-effectiveness analysis, in combination with the sensitivity and specificity of the FLOTAC technique, suggest that the FLOTAC technique can be use in the routine diagnosis of Giardia spp. infection in dogs.

Accurate diagnostic tools for human helminthiasis are crucial for epidemiological surveys, improved patient management, and evaluation of community-based intervention studies. However, the diagnosis of intestinal schistosomiasis and soil-transmitted helminthiasis heavily relies on stool microscopy using the Kato-Katz technique, which has a low sensitivity. The Mini-FLOTAC method is an alternative microscopy-based technique, but its diagnostic performance using sodium acetate-acetic acid-formalin-(SAF)-fixed stool specimens has not been validated. The fixation of stool samples for later examination in a laboratory may reduce logistical and financial barriers of prevalence surveys by not requiring field laboratories. We compared the diagnostic accuracy of the Kato-Katz technique using fresh stool samples with the Mini-FLOTAC technique, employing matched stool samples that were fixed in SAF. Three consecutive stool samples from 149 school-aged children in Côte d'Ivoire were subjected to quintuplicate Kato-Katz thick smears examined on the same day. From the remaining stool, approximately 2g was fixed in 10ml of SAF for about 3 months, and then subjected to the Mini-FLOTAC method, using two flotation solutions (FS2 and FS7). The combined results of multiple Kato-Katz and Mini-FLOTAC readings revealed prevalences of Schistosoma mansoni, Trichuris trichiura and hookworm of 99.3%, 72.5% and 7.4%, respectively. Employing a Bayesian latent class analysis to estimate the true sensitivity of the diagnostic approaches, the sensitivity of Mini-FLOTAC using FS2 was 50.1% (95% Bayesian credible interval (BCI): 30.9-70.2%) for hookworm and 68.0% (95% BCI: 34.9-93.5%) for T. trichiura. When applying Mini-FLOTAC using FS7, the sensitivity was 89.9% (95% BCI: 86.9-97.4%) for S. mansoni, 37.2% (95% BCI: 17.2-60.6%) for hookworm and 67.7% (95% BCI: 33.0-93.0%) for T. trichiura. The specificity ranged from 80.1-95.0% in all Mini-FLOTAC tests. Mini-FLOTAC revealed higher arithmetic mean faecal egg counts (FECs) than the Kato-Katz technique. We found a significant correlation in FECs between Kato-Katz and Mini-FLOTAC for S. mansoni and T. trichiura. We conclude that Mini-FLOTAC shows reasonable diagnostic accuracy when using stool samples fixed in SAF for 3 months, and may be an alternative to multiple Kato-Katz thick smears.

Background Kato-Katz is a widely used method for the diagnosis of soil-transmitted helminth infection. Fecal samples cannot be preserved, and hence, should be processed on the day of collection and examined under a microscope within 60 min of slide preparation. Mini-FLOTAC is a technique that allows examining fixed fecal samples. We assessed the performance of Mini-FLOTAC using formalin-fixed stool samples compared to Kato-Katz and determined the dynamics of prevalence and intensity estimates of soil-transmitted helminth infection over a 31-day time period. Methodology The study was carried out in late 2013 on Pemba Island, Tanzania. Forty-one children were enrolled and stool samples were subjected on the day of collection to a single Kato-Katz thick smear and Mini-FLOTAC examination; 12 aliquots of stool were fixed in 5% formalin and subsequently examined by Mini-FLOTAC up to 31 days after collection. Principal Findings The combined results from Kato-Katz and Mini-FLOTAC revealed that 100% of children were positive for Trichuris trichiura, 85% for Ascaris lumbricoides, and 54% for hookworm. Kato-Katz and Mini-FLOTAC techniques found similar prevalence estimates for A. lumbricoides (85% versus 76%), T. trichiura (98% versus 100%), and hookworm (42% versus 51%). The mean eggs per gram of stool (EPG) according to Kato-Katz and Mini-FLOTAC was 12,075 and 11,679 for A. lumbricoides, 1,074 and 1,592 for T. trichiura, and 255 and 220 for hookworm, respectively. The mean EPG from day 1 to 31 of fixation was stable for A. lumbricoides and T. trichiura, but gradually declined for hookworm, starting at day 15. Conclusions/Significance The findings of our study suggest that for a qualitative diagnosis of soil-transmitted helminth infection, stool samples can be fixed in 5% formalin for at least 30 days. However, for an accurate quantitative diagnosis of hookworm, we suggest a limit of 15 days of preservation. Our results have direct implication for integrating soil-transmitted helminthiasis into transmission assessment surveys for lymphatic filariasis., Author Summary Soil-transmitted helminths are parasitic worms (hookworm, roundworm, and whipworm) that affect hundreds of millions of people. Kato-Katz is the most widely used technique for the diagnosis of soil-transmitted helminth infection. It requires the collection, processing, and microscopic examination of stool samples within the same day. In remote areas, laboratories are often far away from where stool samples are being collected, which makes it difficult to examine the samples on the same day. Mini-FLOTAC is an alternative to Kato-Katz technique that allows analysis of fixed stool samples several days after collection. We assessed the accuracy of the Mini-FLOTAC with fixed stool samples. The study was carried out in late 2013 on Pemba Island and 41 children participated. Fresh stool samples were first examined by Kato-Katz and then fixed in formalin and examined by Mini-FLOTAC (12 examinations within one month). We found that for a qualitative diagnosis (presence or absence of infection) stool samples can be fixed for 31 days. However, for an accurate quantitative diagnosis of hookworm eggs, stool samples should not be stored for more than 15 days, as egg counts decline. Our results have implications for integrating soil-transmitted helminth surveys with other neglected tropical diseases.

Little information is available on the occurrence of endoparasites in pet guinea pigs (Cavia porcellus). The purpose of this study was to evaluate the prevalence of intestinal parasites in cavies kept as pets in southern Italy. Fresh fecal samples were randomly collected from 60 guinea pigs housed in pet shops or privately owned. All fecal samples were processed using the FLOTAC pellet technique to identify and count helminthic eggs/larvae and protozoan cysts/oocysts. In addition, the specimens were analyzed also by the Remel Xpect® Giardia/Cryptosporidium immunoassay. Intestinal parasites were detected in 19 out of 60 guinea pigs (31.7 %). Paraspidodera uncinata eggs were found in 13.3 % (8/60) of the rodents examined, Nippostrongylus-like eggs in 10 % (6/60), and finally Eimeria caviae oocysts were found in 10 % (6/60) of the animals. In one case, both E. caviae oocysts and P. uncinata eggs were found. None of the samples was positive for Cryptosporidium or Giardia. To the authors’ knowledge, this is the first survey of endoparasites in pet guinea pigs in Italy.

Background Helminths and protozoa infections pose a great burden especially in developing countries, due to morbidity caused by both acute and chronic infection. The aim of our survey was to analyze the intestinal parasitic burden in communities from Mwanza region, Tanzania. Methods Subjects (n = 251) from four villages on the South of Lake Victoria have been analyzed for intestinal parasites with direct smear (DS), formol-ether concentration method (FECM) and the newly developed Mini-FLOTAC technique; urinary schistosomiasis was also assessed in a subsample (n = 151); symptoms were registered and correlation between clinic and infections was calculated by chi-squared test and logistical regression. Results Out of the subjects screened for intestinal and for urinary parasites, 87% (218/251) were found positive for any infection, 69% (174/251) carried a helminthic and 67% (167/251) a protozoan infection, almost half of them had a double or triple infection. The most common helminths were hookworms, followed by Schistosoma mansoni and Schistosoma haematobium. Among protozoa, the most common was Entamoeba coli followed by Entamoeba histolytica/dispar and Giardia intestinalis. Mini-FLOTAC detected a number of helminth infections (61.7%) higher than FECM (38.6%) and DS (17.9%). Some positive associations with abdominal symptoms were found and previous treatment was negatively correlated with infection. Conclusion Despite the limited size of the examined population the current study indicates a high prevalence of intestinal parasitic infection in Bukumbi area, Tanzania, and Mini-FLOTAC showed to be a promising diagnostic tool for helminth infections. This high parasitic burden calls for starting a regular deworming programme and other preventive interventions in schools and in the community.

Endoparasites are infrequently reported in ferrets. A cross-sectional survey was conducted to determine the prevalence of intestinal parasites in pet ferrets in southern Italy. Fresh fecal samples were randomly collected from 50 ferrets housed in pet shops or privately owned. All fecal samples were processed using the FLOTAC pellet technique to identify and count helminthic eggs/larvae and protozoan cysts/oocysts. In addition, the samples were analyzed also by the Remel Xpect®Giardia/Cryptosporidium immunoassay. Intestinal parasites were detected in 15 out of 50 ferrets (30%). Eggs of ancylostomids were found in 28.0% (14/50) of the animals and oocysts of Sarcocystis were detected in one ferret (2.0%). None of the samples was positive for Cryptosporidium or Giardia. To the authors’ knowledge, this is the first report of sarcosporidiosis in a pet ferret in Italy.

The present study investigated the occurrence of endoparasites in pet squirrels in southern Italy. Fresh fecal samples were collected from 50 asymptomatic pet squirrels belonging to five different species (Callosciurus finlaysonii, n=6, C. prevosti, n=6; Tamias striatus, n=26, T. sibiricus, n=10; Sciurus carolinensis, n=2) housed both in pet shops and/or in private residences. All fecal samples were processed using the FLOTAC pellet technique to identify and count helminth eggs/larvae and protozoan cysts/oocysts. In addition, to detect Cryptosporidium spp. and Giardia spp. the samples were analyzed by the Remel Xpect(®) immunoassay. Helminth eggs were detected in 9 out of 50 squirrels. Specifically, eggs of Dicrocoelium dendriticum were found in 5 squirrels (C. finlaysonii, n=2; C. prevosti, n=2; T. striatus, n=1); eggs of the pinworm Syphacia spp. in 3 squirrels (C. prevosti, n=2; T. striatus, n=1); and eggs of gastrointestinal nematoda (Nippostrongylus-like) were found in 1 subject (C. prevosti). Finally, two squirrels (C. prevosti) had multiple parasitic infections with D. dendriticum and Capillaria hepatica, and with D. dendriticum and Strongyloides spp., respectively. None of the samples were positive for Cryptosporidium spp. or Giardia spp. or any other protozoa (e.g. Eimeria). To the authors' knowledge, this is the first report of a D. dendriticum natural infection in pet rodents.

Background Helminths and protozoa infections pose a great burden especially in developing, countries, due to morbidity caused both by acute and chronic infections. Data on distribution of intestinal parasitic infections among the native and expatriates populations in Himachal Pradesh are scarce. The aim of our survey was to analyze the intestinal parasitic burden in communities from Dharamsala, Kangra district, in clinical and public health settings. We also field-tested the mini- FLOTAC, an innovative diagnostic device. Methods Subjects referring to the Tibetan Delek Hospital for abdominal discomfort and all children of the Tibetan Primary School in Dharamsala were screened for intestinal parasitic infections with direct smear, formol-ether concentration (FEC) method and mini-FLOTAC, their clinical history was recorded, and correlations between clinical symptoms and infections analyzed. Results 152 subjects were screened for intestinal parasites, of which 72 subjects in the outpatients department (OPD) (36 expatriates and 36 natives) and 80 in the school. 60% of schoolchildren and 57% of OPD patients were found positive for any infection, the most represented were protozoa infections (50%), whereas helminthic infections accounted only for 13% and 20% in OPD patients and schoolchildren, respectively. The most prevalent among helminths was Ascaris lumbricoides (11%). Giardia intestinalis was more present among schoolchildren than the OPD patients (20% vs 6%) and E. histolytica / dispar was more prevalent among the OPD patients (42%) than the school children (23%). Correlations were found between nausea and loose or watery stools and parasitic infections, particularly in expatriates, whereas schoolchildren, despite being as infected as adults, were completely asymptomatic. Mini-FLOTAC detected higher number of helminth infections whereas FEC method was more accurate for the diagnosis of protozoa. Conclusions This study presents an accurate snapshot of intestinal parasitic infections in Dharamsala, and their high prevalence calls for more awareness and control measures. Mini-FLOTAC is a promising and simple technique for the diagnosis of helminth infections.

Background Soil-transmitted helminths and intestinal protozoa infection are widespread in developing countries, yet an accurate diagnosis is rarely performed. The aim of this study was to evaluate the recently developed mini–FLOTAC method and to compare with currently more widely used techniques for the diagnosis of intestinal parasitic infections in different settings. Methodology/Principal Findings The study was carried out in Dharamsala, Himachal Pradesh, India, and in Bukumbi, Tanzania. A total of 180 pupils from two primary schools had their stool analyzed (n = 80 in Dharamsala and n = 100 in Bukumbi) for intestinal parasitic infections with three diagnostic methods: direct fecal smear, formol-ether concentration method (FECM) and mini-FLOTAC. Overall, 72% of the pupils were positive for any intestinal parasitic infection, 24% carried dual infections and 11% three infections or more. The most frequently encountered intestinal parasites were Entamoeba coli, Entamoeba histolytica/dispar, Giardia intestinalis, hookworm, (and Schistosoma mansoni, in Tanzania). Statistically significant differences were found in the detection of parasitic infections among the three methods: mini-FLOTAC was the most sensitive method for helminth infections (90% mini-FLOTAC, 60% FECM, and 30% direct fecal smear), whereas FECM was most sensitive for intestinal protozoa infections (88% FECM, 70% direct fecal smear, and 68% mini-FLOTAC). Conclusion/Significance We present the first experiences with the mini-FLOTAC for the diagnosis of intestinal helminths and protozoa. Our results suggest that it is a valid, sensitive and potentially low-cost alternative technique that could be used in resource-limited settings — particularly for helminth diagnosis., Author Summary The mini-FLOTAC has been recently developed as a novel direct method for the diagnosis of intestinal parasitic infections. Mini-FLOTAC attempts to address the challenge of using modern technology matched with high sensitivity, affordability, and appropriateness of diagnosis in resource-limited settings where intestinal parasitic infections are widespread. We compared accuracy and feasibility of mini-FLOTAC with currently more widely used diagnostic methods, such as the direct fecal smear and the formol-ether concentration method. Our study was carried out in Dharamsala, India, and in Bukumbi, Tanzania in order to evaluate the methods under different field conditions and diverse parasitic infection profiles. Among 180 primary schoolchildren examined, 72% were positive for any intestinal parasitic infection. Mini-FLOTAC detected the highest number of helminth infections (90% sensitivity), whereas the formol-ether concentration was the most sensitive approach for intestinal protozoa infections (88% sensitivity). The logistic advantages of mini-FLOTAC are that the procedure does not require any centrifugation step or expensive equipment, it can be performed on fresh and fixed stool samples, and only requires 10–12 min of preparation before microscopic analysis. Our data suggest that mini-FLOTAC is a promising diagnostic tool for helminth diagnosis; therefore, follow-up studies in other settings are warranted.