Toward automating the diagnosis of gastrointestinal parasites in cats and dogs.

Diagnosing gastrointestinal parasites by microscopy slide examination often leads to human interpretation errors, which may occur due to fatigue, lack of training and infrastructure, presence of artifacts (e.g., various types of cells, algae, yeasts), and other reasons. We have investigated the stages in automating the process to cope with the interpretation errors. This work presents advances in two stages focused on gastrointestinal parasites of cats and dogs: a new parasitological processing technique, named TF-Test VetPet, and a microscopy image analysis pipeline based on deep learning methods. TF-Test VetPet improves image quality by reducing cluttering (i.e., eliminating artifacts), which favors automated image analysis. The proposed pipeline can identify three species of parasites in cats and five in dogs, distinguishing them from fecal impurities with an average accuracy of 98,6%. We also make available the two datasets with images of parasites of dogs and cats, which were obtained by processing fecal smears with temporary staining using TF-Test VetPet.

Detection of intestinal parasites in human faecal samples using dissolved air flotation.

OBJECTIVE: Ova and parasite (O&P) examination is recommended for the laboratory diagnosis of agents causing parasitic infections; however, this exam requires scientific and technological improvements to enhance its diagnostic validity. Dissolved air flotation (DAF) is an efficient technical principle separating suspended solids in a liquid medium. We aimed to develop and validate a new procedure for intestinal parasite detection with DAF. METHODS: In this study, we collected samples from 500 volunteers, screened them by direct examination, and transferred the material to tubes using the Three Faecal Test (TF-Test) for triplicate DAF tests. We evaluated physical-chemical parameters and DAF prototype components through quantifying parasites recovered from floated and non-floated regions of the flotation column. The DAF operation protocol was validated with the gold standard results. RESULTS: The 10% saturated volume proportion and cationic surfactant showed regularity and high parasite recovery (80%). Modifications of the needle device did not influence parasite recovery (p > 0.05). Sensitivity, specificity, accuracy and kappa agreement obtained with the DAF protocol were 91%, 100%, 93% and substantial (k = 0.64), respectively. CONCLUSION: The DAF principle could be used to process faecal samples in routine laboratory exams, enabling intestinal parasite detection.

Dissolved air flotation as potential new mechanism for intestinal parasite diagnosis in feces.

The parasitological examination of feces is recommended for the laboratory diagnosis of intestinal parasites due to its practicality, low-cost, and moderate diagnostic sensitivity. Dissolved Air Flotation (DAF) is an efficient technical principle used in other areas to separate dispersed solids. This study sought the preliminary evaluation of a proof-of-concept prototype as a tool for detecting species of parasites by adjusting DAF. Two DAF prototype units were developed to evaluate microbubbles' generation amidst fecal suspension and parasites' capture. For this evaluation, samples were screened and processed by the TF-Test technique (Control) and simultaneously by DAF device. The dimensional and attachment characteristics in the formation of the microbubbles were evaluated, and the percentage of parasitic recovery in floated and not-floated regions compared by Student's t-test. The second prototype unit proved to be more efficient in forming microbubbles with diameters between 34 and 170µm. The flotation tests showed a recovery of 73.27%, 58.12%, 37.85%, and 91.89% for Ascaris lumbricoides, Hymenolepis diminuta, Giardia duodenalis, and Strongyloides stercoralis, respectively. This study confirmed the selective interaction between microbubbles and parasite eggs and larvae during the flotation process using the DAF principle for the first time through imaging.

A historical review of the techniques of recovery of parasites for their detection in human stools.

Since the early 20th century, the detection of intestinal parasites has improved with the development of several techniques for parasitic structures recovery and identification, which differ in sensitivity, specificity, practicality, cost, and infrastructure demand. This study aims to review, in chronological order, the stool examination techniques and discuss their advantages, limitations, and perspectives, and to provide professionals and specialists in this field with data that lays a foundation for critical analysis on the use of such procedures. The concentration procedures that constitute the main techniques applied in routine research and in parasitological kits are a) spontaneous sedimentation; b) centrifugation-sedimentation with formalin-ethyl acetate; and c) flotation with zinc sulfate solution. While selecting a technique, one should consider the purpose of its application and the technical-operational, biological, and physicochemical factors inherent in the procedures used in stool processing, which may restrict its use. These intrinsic limitations may have undergone procedural changes driven by scientific and technological development and by development of alternative methods, which now contribute to the improvement of diagnostic accuracy.

A historical review of the techniques of recovery of parasites for their detection in human stools

Abstract Since the early 20th century, the detection of intestinal parasites has improved with the development of several techniques for parasitic structures recovery and identification, which differ in sensitivity, specificity, practicality, cost, and infrastructure demand. This study aims to review, in chronological order, the stool examination techniques and discuss their advantages, limitations, and perspectives, and to provide professionals and specialists in this field with data that lays a foundation for critical analysis on the use of such procedures. The concentration procedures that constitute the main techniques applied in routine research and in parasitological kits are a) spontaneous sedimentation; b) centrifugation-sedimentation with formalin-ethyl acetate; and c) flotation with zinc sulfate solution. While selecting a technique, one should consider the purpose of its application and the technical-operational, biological, and physicochemical factors inherent in the procedures used in stool processing, which may restrict its use. These intrinsic limitations may have undergone procedural changes driven by scientific and technological development and by development of alternative methods, which now contribute to the improvement of diagnostic accuracy.

Use of the aqueous biphasic system as an alternative for concentration of Ascaris lumbricoides eggs, with non-toxic separation of faecal residues and fats.

OBJECTIVES: Human enteroparasites are considered a serious public health problem in underdeveloped countries located in world regions with tropical, subtropical and equatorial climates. These parasites are commonly diagnosed by the Parasitological Examination of Faeces (PEF), performed by conventional techniques and/or commercial kits that result in tests with low-to-moderate sensitivity, due to the use of destructive chemical solvents to parasite structures, and to present excess adipose substance and digestive residues in their microscopic slides. In order to improve the efficacy of these tests/examinations, this study aimed to investigate a new alternative for the PEF, with the use of Aqueous Biphasic System (ABS). METHODS: For this, four ABSs containing poly (ethylene glycol), PEG (PEG-4000 and PEG-6000), dipotassium phosphate and sodium citrate at different concentrations in the biphasic systems were evaluated with faecal samples containing eggs of Ascaris lumbricoides. RESULTS: The ABS consisting of PEG-4000 and dipotassium phosphate, at concentrations of 55% w/w and 20% w/w, respectively, achieved 100% satisfactory results compared to the conventional TF-Test technique in terms of preservation and concentration of A. lumbricoides eggs, with adequate separation of digestive residues, without using a centrifuge or chemical solvents that may cause harm to the parasites. CONCLUSIONS: This study presents ABS as a new low-cost technical principle for the detection of parasite eggs in PEF. The new technique is simple, fast, non-toxic, not harmful to the parasite and does not require a centrifuge.

UTILISATION DU SYSTÈME BIPHASIQUE AQUEUX COMME ALTERNATIVE À LA CONCENTRATION DES ŒUFS D'ASCARIS LUMBRICOIDES, AVEC SÉPARATION NON TOXIQUE DES RÉSIDUS FÉCAUX ET DES GRAISSES: OBJECTIFS: Les entéroparasites humains sont considérés comme un sérieux problème de santé publique dans les pays sous-développés situés dans des régions du monde aux climats tropicaux, subtropicaux et équatoriaux. L'examen parasitologique des matières fécales (EPMF) permet généralement de diagnostiquer ces parasites, à l'aide de techniques classiques et/ou de kits commerciaux, conduisant à des tests de sensibilité faible à modérée, dus à l'utilisation de solvants chimiques destructeurs pour les structures du parasite et résultant en excès de substances adipeuses et de résidus digestifs dans leurs lames microscopiques. Afin d'améliorer l'efficacité de ces tests/examens, cette étude visait à étudier une nouvelle alternative à l'EPMF, avec l'utilisation du Système Biphasique Aqueux (SBA). MÉTHODES: Pour cela, quatre SBA contenant du poly (éthylène glycol), du PEG (PEG-4000 et PEG-6000), du phosphate dipotassique et du citrate de sodium à différentes concentrations ont été évalués avec des échantillons de matières fécales contenant des œufs d'Ascaris lumbricoides. RÉSULTATS: Le SBA constitué de PEG-4000 et de phosphate dipotassique, à des concentrations respectives de 55% poids/poids et 20% poids/poids, a obtenu des résultats satisfaisants à 100% par rapport à la technique conventionnelle du TF-Test, en termes de préservation et de concentration des œufs de A. lumbricoides, avec une séparation adéquate des résidus digestifs, sans utiliser de centrifugeuse ni de solvants chimiques pouvant détruire les parasites. CONCLUSIONS: Cette étude présente le SBA comme un nouveau principe technique peu coûteux pour la détection des œufs de parasites dans l'EPMF. La nouvelle technique est simple, rapide, non toxique, n'endommage pas le parasite et ne nécessite pas de centrifugeuse.