Eggs In Faecal Samples Research Articles

Gastrointestinal Helminths in Local (Black Bengal) and Jamunapari Goats of Barishal Sadar, Southern Bangladesh

Gastrointestinal helminths are important causes of hindering global goat production. To find the prevalence of gastrointestinal helminths of Black Bengal and Jamunapari breeds of goats, the current investigation was carried out at Barishal Sadar Upazilla of Barishal district, Bangladesh. The gastrointestinal helminths were identified through coprological examination. A total of 112 fecal samples were collected from household goats across different seasons, breeds, sexes, and ages. During the study period, four types of gastrointestinal helminths were identified based on the presence of helminth eggs in fecal samples. The overall prevalence of gastrointestinal helminths in goats was found to be 82.1%, while the prevalence rates of Fasciola gigantica (F. gigantica), Paramphistomum spp., Bunostomum spp., and Hemonchus spp. were 34.8% (95% CI: 1.4-2.5), 22.3% (95% CI: 0.7-1.8), 14.3% (95% CI: 0.1-1.5), and 10.7% (95% CI: 0.2-1.0), respectively. A significantly different prevalence was observed among different gastrointestinal helminths in goats. A significantly lower prevalence of F. gigantica and Paramphistomum spp. was observed in male goats, compared to females. A higher prevalence of F. gigantica was significantly observed during the winter, compared to the summer. The current study elucidates that F. gigantica was more prevalent in female goats. The current study indicated that F. gigantica was more prevalent in female goats. These findings underscore the importance of further research and control measures to manage gastrointestinal helminth infections in goats across southern Bangladesh and other regions with similar environmental conditions.

Coproscopical diagnosis of patent Fasciola hepatica infections in sheep – A comparison between standard sedimentation, FLUKEFINDER® and a combination of both

The liver fluke Fasciola hepatica is a highly pathogenic and zoonotic trematode with a cosmopolitan distribution. In livestock, infections may lead to significant economic losses if not diagnosed promptly and treated effectively. Particularly for small ruminants, the standard method for the detection of fluke infection is based on coproscopical methods such as the sedimentation method, which detects F. hepatica eggs in faecal samples. In this respect a recent innovative coproscopical approach to diagnose patent infections is the FLUKEFINDER® method, which relies on differential sieving before sedimentation. These two methods and a combination of both methods that allows larger amounts of faeces to be processed with the FLUKEFINDER® apparatus were compared, to assess which method is most appropriate to determine the prevalence and intensity of F. hepatica egg shedding. The methods were compared for their ability to recover eggs from ovine faecal samples containing different numbers of fluke eggs per gram (EPG) of faeces and diluting the samples further by mixing with faeces from uninfected sheep. To compare the specificity of the test procedures, positive and negative samples with a low EPG were analysed in parallel by an investigator blinded to the nature of the samples. Significant differences concerning the EPG outcome were found: The FLUKEFINDER® method demonstrated the highest EPG values (p < 0.001) in the undiluted samples as well as in all mixing levels, followed by the modified FLUKEFINDER® method. The standard sedimentation showed the lowest EPG values and the highest variability between technical replicates. The precision of the FLUKEFINDER® method and the modified FLUKEFINDER® method were significantly higher than the precision of the standard sedimentation as determined by comparison of variability between technical replicates. The highest raw egg counts were detected using the modified FLUKEFINDER® method. The FLUKEFINDER® method and the combined method showed a sensitivity of 100 % even at the lowest egg concentrations, whereas the sensitivity of the standard sedimentation was 98.1 % for the same set of samples (i.e. one false negative sample). In a separate investigation aiming to estimate the specificity no differences were found between the three methods: all protocols showed 100 % specificity and were able to correctly distinguish between truly positive and truly negative samples without any evidence of cross-contamination between positive and negative samples processed in parallel.

Feasibility of a DNA biosensor assay based on loop-mediated isothermal amplification combined with a lateral flow dipstick assay for the visual detection of Ascaridia galli eggs in faecal samples

ABSTRACT Ascaridia galli is an important nematode that causes ascaridiasis in free-range and indoor system chicken farms. Infection with A. galli may damage the intestinal mucosa and inhibit nutrient absorption, leading to a reduced growth rate, weight loss and a decreased egg production. Consequently, A. galli infection is a significant health problem in chickens. In this study, we developed a loop-mediated isothermal amplification coupled with a lateral flow dipstick (LAMP-LFD) assay for the visual detection of A. galli eggs in faecal samples. The LAMP-LFD assay consists of six primers and one DNA probe that recognize the internal transcribed spacer 2 (ITS2) region; it can be performed within 70 min and the results can be interpreted with the naked eye. Using the LAMP-LFD assay developed in this study, A. galli DNA was specifically amplified without any cross-reactions with other related parasites (Heterakis gallinarum, Raillietina echinobothrida, R. tetragona, R. cesticillus, Cotugnia sp., Echinostoma miyagawai) and definitive hosts (Gallus gallus domesticus, Anas platyrhynchos domesticus). The minimum detectable DNA concentration was 5 pg/μl, and the detectable egg count was 50 eggs per reaction. The assay can be performed in a water bath, without the need for post-mortem morphological investigations and laboratory instruments. It is therefore a viable alternative for the detection of A. galli in chicken faeces and can replace classical methods in field screening for epidemiological investigations, veterinary health and poultry farming management. RESEARCH HIGHLIGHTS This is the first study using the LAMP-LFD assay for Ascaridia galli detection. The results can be observed by the naked eye. The developed assay can be used to detect Ascaridia galli eggs in faecal samples.

The Use of MALDI-TOF MS as a Diagnostic Tool for Adult Trichuris Species.

Trichuriasis is considered a neglected tropical disease, being the second most common helminthiasis in humans. Detection of Trichuris in routine diagnosis is usually done by microscopic detection of eggs in fecal samples. Other molecular analyses are more reliable and could be used, but these analyses are not routinely available in clinical microbiology laboratories. The use of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is increasing since the last decades due to its recent evidence as a potential role for reliable identification of microorganisms and a few nematodes. But, for parasites detection, normalized protocols and the acquisition and introduction of new species to the database are required. We carried out a preliminary study confirming the usefulness of MALDI-TOF MS for the rapid and reliable identification of Trichuris suis used as control and the creation of an internal database. To create main spectra profiles (MSPs), the different parts of five whipworms (esophagus and intestine) were used, developing different tests to verify the repeatability and reproducibility of the spectra. Thus, to validate the new internal database, 20 whipworms, separating the esophagus and intestine, were used, of which 100% were accurately identified as T. suis, but could not distinguish between both parts of the worm. Log score values ranged between 1.84 and 2.36, meaning a high-quality identification. The results confirmed that MALDI-TOF MS was able to identify Trichuris species. Additionally, a MALDI-TOF MS profile of T. suis proteome was carried out to develop the first internal database of spectra for the diagnosis of trichuriasis and other Trichuris spp.

Assessment of three DNA extraction kits for the absolute quantification of strongyle nematode eggs in faecal samples

BackgroundHaemonchus contortus is one of the most pathogenic gastrointestinal nematodes of small ruminants. The current diagnostic approach for the detection of this species relies on coproscopic methods, which both have low sensitivity and are time consuming. Methods employing detection through DNA amplification, such as droplet digital polymerase chain reaction (ddPCR), offer an advantageous approach to the diagnosis of H. contortus. However, DNA extraction protocols need to be constantly updated for the optimal retrieval of diagnostically usable template. Here, we describe the evaluation of three genomic DNA extraction kits for the detection and quantification of H. contortus ITS2 amplicon DNA from faecal samples, using droplet digital PCR.ResultsDNA samples, extracted from faecal material with the Nucleospin DNA Stool kit, produced the highest amounts of ITS2 amplicon copies and had the lowest coefficient of variation across different dilutions and sample types (fresh or frozen) out of the tested kits (Nucleospin DNA Stool, E.Z.N.A.® Stool DNA Kit and QIAamp Fast DNA Stool Mini Kit). Furthermore, the protocol of this kit has the fewest number of steps and the price of DNA extraction per sample is reasonable (2.77 €).ConclusionsThe Nucleospin DNA Stool kit is an attractive option for the detection and quantification of H. contortus DNA in faecal samples of small ruminants in a diagnostic setting.

A Schistosoma mansoni tri- and tetramer microsatellite catalog for genetic population diversity and differentiation

All Schistosoma mansoni tri- and tetranucleotide repeat microsatellites published as of December 2018 were identified. All 52 were evaluated for autosomal location, strength of amplification, scorability and behavior as single-copy loci by polyacrylamide and capillary gel electrophoresis. Of these, 27 were unique, autosomal, polymorphic, easily scored and single copy as assessed on pooled adult worm DNA from two different continental origins and adult worm clones. These microsatellites were distributed across all seven autosomal chromosomes. On laboratory strains their heterozygosity ranged from 0.22 to 0.77. Individual markers had 5–13 alleles, allelic richness of 2–10 and an effective allele number of 1.3–8.14. Those infected by Schistosoma mansoni carry many genetically distinct, sexually reproducing parasites, therefore, for an individual infection the complete allele frequency profile of their progeny consists of a pool of DNA from multiple diploid eggs. Using a set of 25 microsatellites, we calculated allele frequency profiles of eggs in fecal samples from people in two Brazilian communities separated by 6 km: Jenipapo (n = 80) and Volta do Rio (n = 38). There were no a priori characteristics that could predict the performance of markers in natural infections based on their performance with laboratory strains. Increasing marker number did not change accuracy for differentiation and diversity but did improve precision. Our data suggest that using a random set of 10–20 microsatellites appears to result in values that exhibit low standard deviations for diversity and differentiation indices. All identified microsatellites as well as PCR conditions, allele size, primer sequences and references for all tri- and tetramer microsatellites markers presented in this work are available at: https://sites.google.com/case.edu/cwru-and-fiocruz-wdrc/home.

Risk Factors for Clonorchis sinensis Infection in Residents of Binyang, Guangxi: A Cross-Sectional and Logistic Analysis Study.

Background: Clonorchiasis is a serious food-borne parasitic disease caused by Clonorchis sinensis infection. C. sinensis, a major fish-borne trematode, is a known causative agent of cholangiocarcinoma. The risk factors for C. sinensis infection include individual eating behaviors and environmental factors. In this study, we evaluated the C. sinensis infection rate and the associated risk factors among residents in Binyang County, Guangxi, China.Methods: In 2016 and 2017, five villages from Binyang, Guangxi were selected by multistage cluster random sampling for a cross-sectional study. A modified Kato-Katz thick smear method was used to examine C. sinensis eggs in fecal samples in triplicate (three smears for each sample). Both uni-variate and multi-variate logistic regression analyses were carried out to identify the risk factors for C. sinensis infection.Results: A total of 1,977 fecal samples were collected from villagers in the investigated areas. The overall infection rates of C. sinensis in Binyang County was 20.49% (405/1,977). The mean age of participants was 39.42 ± 23.48 (range: 3–89 years old), and the highest infection rate (33.72%) was seen in the age group of 40-49 years old, followed by those aged 50–59 (31.83%). Multi-variate logistic regression analysis showed that higher infection rates were significantly associated with males (aOR = 6.51, 95% CI = 4.67–9.08), Zhuang (aOR = 2.41, 95% CI = 1.62–3.59), ages (aOR = 33.51, 95% CI = 10.13–110.86), frequency of raw fresh fish consumption (aOR = 14.56, 95% CI = 9.80–21.63), and close contact with cats and dogs (aOR = 1.53, 95% CI = 1.02–2.30). Occupations and education levels showed no significant association with C. sinensis infection (P > 0.05).Conclusions: High levels of C. sinensis infection were observed among residents in Binyang County, Guangxi. Intervention strategies should be strengthened among the investigated population at high risk, such as males, Zhuang and older individuals, especially those who frequently eat raw fresh fish. In addition, the individuals contacting with cats and/or dogs were observed to have significantly higher infection rate of C. sinensis than those having no contact with cats and dogs. The association between contacting with cats and/or dogs and C. sinensis infection needs to be explored and confirmed in the future study by more epidemiological investigations of human C. sinensis infection from different areas.

Lungworms (Metastrongylus spp.) and intestinal parasitic stages of two separated Swiss wild boar populations north and south of the Alps: Similar parasite spectrum with regional idiosyncrasies.

Metastrongylus sp. is a lungworm with worldwide distribution in wild boars and other suids. In Switzerland, two wild boar populations are geographically divided by the Alps. We investigated 84 wild boars, 52 from north and 32 from south of the Alps, different in sex and age (juveniles, subadults, adults), shot between September and December 2018. The lungs were macroscopically examined and dissected for the detection of Metastrongylus specimens. Additionally, faecal samples were obtained from 55 animals and analysed by sedimentation/flotation and the mini-FLOTAC® method. Overall, 12′774 Metastrongylus sp. specimens were isolated: prevalence was 77.4% and mean burden 196.5 (range: 1–2589), with no significant differences between north (80.8%, 218.0) and south (71.9%, 157.4) nor between sexes and age groups. Macroscopically, dense nodular lesions associated with Metastrongylus sp. were present in 19 out of 65 (33.9%) positive lungs. Five Metastrongylus sp. were detected: M. pudendotectus (67.9%), M. salmi (63.1%), M. confusus (56%), M. apri (44%) and M. asymmetricus (17.9%), with a significant difference (p = 0.012) between north (32.7%) and south (62.5%) for M. apri. The lungworm population was female biased. The number of Metastrongylus sp. eggs in faecal samples did not correlate with worm burdens. Furthermore, the following endoparasites were detected: Isospora suis/Eimeria sp. (74.5%), Strongyloides suis (27.3%%), Trichuris suis (20.0%), Hyostrongylus rubidus/Oesophagostomum sp. (18.2%), Globocephalus sp. (9.1%), Capillaria sp. (7.3%), Ascaris suum (3.6%), Giardia sp. (3.6%) and Balantidium coli (1.8%), with significant differences for S. suis (north 36.1% > south 10.5%) and Globocephalus sp. (only south, 26.3%). Although geographically separated, both Swiss wild boar populations share similar parasite spectra, while also showing some regional idiosyncrasies partially explained by ecological and climatic factors. Despite their clinical relevance being unknown, accurate knowledge concerning the distribution of endoparasites in the wildlife reservoir is relevant to better understand risk factors for the domestic pig population.

Prevalence and Intensity of Parasitic Worms on Free-Range Chickens and Egg-Laying Chickens in Muara Badak sub-District, Kutai Kartanegara

One of the diseases often threatens the health of free-range chickens and egg-laying chickens are worm parasites (endoparasites). Parasites can be transmitted through food, drinks and the cage environment. Sanitation is one of the preventive steps carry out by farmers to protect livestock against infection from the environment. The objective of study was to determine the prevalence and intensity of parasitic worm eggs that infect the digestive tracts of free-range chickens and egg- laying hens in Muara Badak District. Research was done by observing worm eggs in fecal samples. A total of 60 samples consisting of 30 samples of egg-laying hens in Tanjung Village and 30 samples of free-range chickens in Badak Baru Village, Muara Badak. For the sample analysis technique used 2 methods are namely the floating method and the sedimentation method. The results has showed the type of worm eggs are infected the digestive tract found in free-range chickens and egg-laying chickens had consisted of Echinostoma revolutum, Raillietina echinobothrida, Raillietina tetragona, Davainea proglotina, Amoebotaenia sphenoides, Ascaridia galli, Capillaria annulata and Trichostrongylus tenuis. The highest prevalence in free- range chickens and egg-laying chickens were found in Ascaridia galli was 53.33% and 40%. While the lowest prevalence in free-range chickens against Raillietina echinobothrida and Amoebotaenia sphenoides 3.33%. egg-laying chickens are against Echinostoma revolutum, Raillietina tetragona and Trichostrongylus tenuis 6.67%. The highest intensity in free-range chickens and against Ascaridia galli was 141 eggs/individual and in egg-laying chickens was 139 eggs/individuals. The lowest intensity in this research was found in free-range chickens against Amoebotaenia sphenoides and Raillietina echinobothrida 3 eggs/individuals and egg-laying chickens against Echinostoma revolutum 5 eggs/individuals. Keywords: Endoparasites, Prevalence, Intensity, Chicken, hen.

Identification of Adult Fasciola spp. Using Matrix-Assisted Laser/Desorption Ionization Time-of-Flight (MALDI-TOF) Mass Spectrometry

Fascioliasis is a neglected trematode infection caused by Fasciola gigantica and Fasciola hepatica. Routine diagnosis of fascioliasis relies on macroscopic identification of adult worms in liver tissue of slaughtered animals, and microscopic detection of eggs in fecal samples of animals and humans. However, the diagnostic accuracy of morphological techniques and stool microscopy is low. Molecular diagnostics (e.g., polymerase chain reaction (PCR)) are more reliable, but these techniques are not routinely available in clinical microbiology laboratories. Matrix-assisted laser/desorption ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) is a widely-used technique for identification of bacteria and fungi; yet, standardized protocols and databases for parasite detection need to be developed. The purpose of this study was to develop and validate an in-house database for Fasciola species-specific identification. To achieve this goal, the posterior parts of seven adult F. gigantica and one adult F. hepatica were processed and subjected to MALDI-TOF MS to create main spectra profiles (MSPs). Repeatability and reproducibility tests were performed to develop the database. A principal component analysis revealed significant differences between the spectra of F. gigantica and F. hepatica. Subsequently, 78 Fasciola samples were analyzed by MALDI-TOF MS using the previously developed database, out of which 98.7% (n = 74) and 100% (n = 3) were correctly identified as F. gigantica and F. hepatica, respectively. Log score values ranged between 1.73 and 2.23, thus indicating a reliable identification. We conclude that MALDI-TOF MS can provide species-specific identification of medically relevant liver flukes.

Epidemiological aspects of Ancylostoma spp. infection in naturally infected dogs from São Paulo state, Brazil.

Ancylostoma spp. are the most prevalent canine nematode parasites in Brazil. Despite their widespread parasitism in juvenile dogs, infections may occur regardless of host age. Although eosinophilia is a frequent finding in experimental infections, it is uncommon in naturally infected dogs. This study aimed to evaluate the prevalence of Ancylostoma spp. and the eosinophil blood counts (EBC) in naturally infected dogs, with or without comorbidities, admitted to the Veterinary Teaching Hospital of the São Paulo State University (UNESP), School of Veterinary Medicine and Animal Science, Botucatu campus, São Paulo state, Brazil, from 2009 to 2018. All retrospective data were gathered from veterinary medical records (VMR); diagnosis of Ancylostoma infection required the identification of eggs in fecal samples by the simple flotation test (SFT). Ancylostoma-infected animals were evaluated for other intestinal parasitic coinfections (IPC) by either the SFT or the centrifugal-flotation test. Dogs free of any gastro-intestinal parasites were prospectively included in control group (Group C). Ancylostoma-infected animals were defined: Ancylostoma spp. only intestinal parasite infection (Group A), Ancyslostoma spp. with concurrent IPC (Group B), Ancylostoma spp. only intestinal parasite infection with concurrent systemic disorders (Group D), and Ancylostoma spp. with both IPC and concurrent systemic disorders (Group E). The overall prevalence of Ancylostoma spp. was 12.1% (207/1715), that was decreased from 2014 to 2018 (9.7%) relative to the 2009-to-2013 period (13.9%). Prevalence was not significantly different between dogs <1-year-old (10.7% [51/478]) and≥1year-old (11.7% [130/1109]). IPC was observed in 45.4% (93/205) of the animals positive for Ancylostoma spp., while dogs <1year old experienced IPC more often (58.8% [30/51]) than dogs ≥1year old (38.5% [50/130]) (P=0.02). Group A (n=35) exhibited median EBC of 1.05×109/L, and an eosinophilia ratio of 34.3% that was significantly higher (P<0.05) than Group C (0.45×109/L and 4.1%, respectively). Both variables did not differ in Group B (n=20), D (n=39) or E (n=36) in comparison to Group C (P>0.05). By ROC curve analysis, only Group A generated a significant area under the curve (0.72). With EBC cutoff of 0.85×109 eosinophils/L, sensitivity and specificity were 65.7% and 70.8%, respectively. Eosinophil counts alone may be helpful in raising suspicion of an Ancylostoma spp. infection if further intestinal parasites and concurrent disorders are absent. However, local prevalence data and epidemiological findings should also be evaluated, since eosinophilia is less frequently observed with Ancylostoma spp. infections in the presence of comorbidities.

Diagnostic performance of McMaster, Wisconsin, and automated egg counting techniques for enumeration of equine strongyle eggs in fecal samples

Fecal egg counts are the cornerstone of equine parasite control programs. Previous work led to the development of an automated, image-analysis-based parasite egg counting system. The system has been further developed to include an automated reagent dispenser unit and a custom camera (CC) unit that generates higher resolution images, as well as a particle shape analysis (PSA) algorithm and machine learning (ML) algorithm. The first aim of this study was to conduct a comprehensive comparison of method precision between the original smartphone (SP) unit with the PSA algorithm, CC/PSA, CC/ML, and the traditional McMaster (MM) and Wisconsin (MW) manual techniques. Additionally, a Bayesian analysis was performed to estimate and compare sensitivity and specificity of all five methods. Feces were collected from horses, screened with triplicate Mini-FLOTAC counts, and placed into five categories: negative (no eggs seen), > 0 - ≤ 200 eggs per gram (EPG), > 200 - ≤ 500 EPG, > 500 - ≤ 1000 EPG, and > 1000 EPG. Ten replicates per horse were analyzed for each technique. Technical variability for samples > 200 EPG was significantly higher for MM than CC/PSA and CC/ML (p < 0.0001). Biological variability for samples> 0 was numerically highest for CC/PSA, but with samples > 200 EPG, MM had a significantly lower CV than MW (p = 0.001), MW had a significantly lower CV than CC/PSA (p < 0.0001), CC/ML had a significantly lower CV than both MW and SP/PSA (p < 0.0001, p = 0.0003), and CC/PSA had a significantly lower CV than CC/SP (p = 0.0115). Sensitivity was> 98 % for all five methods with no significant differences. Specificity, however, was significantly the highest for CC/PSA, followed numerically by SP/PSA, MM, CC/ML, and finally MW. Overall, the automated counting system is a promising new development in equine parasitology. Continued refinement to the counting algorithms will help improve precision and specificity, while additional research in areas such as egg loss, analyst variability at the counting step, and accuracy will help create a complete picture of its impact as a new fecal egg count method.

Capillaria boehmi (syn. Eucoleus boehmi): Challenging treatment of a rarely diagnosed nasal nematode in dogs and high prevalence in Swiss foxes

Despite morphological differences of eggs and adults, Capillaria boehmi infections have been occasionally misdiagnosed as C. aerophila infections in the past. Capillaria boehmi is found in the nasal and paranasal sinuses of wild canids and dogs, which may suffer from nasal discharge, sneezing, epistaxis and, importantly, their scent can be impaired. In this study we present three challenging cases of nasal capillariosis in dogs, report and review the variable success of anthelmintic treatments and investigate C. boehmi prevalence in Swiss red foxes, considered as potential wild life reservoir. Out of two females and one male dog (all scent hounds, aged 3–9 years and weighing 19−31 kg), two dogs were previously coproscopically misdiagnosed with Trichuris infections. Two dogs showed clinical signs such as sneezing, coughing and impaired scent. From one dog adult living C. boehmi were obtained by nasal lavage. The identity of worms and eggs of all three dogs were genetically confirmed (18S rRNA, 100 % identity in 578 base pairs). Dogs 1–3 were followed-up for overall 54, 8, and 67 months, respectively. All dogs repeatedly excreted C. boehmi eggs in faecal samples despite treatments with the following compounds, in various dosage and retreatment protocols: fenbendazole, milbemycin oxime (orally), moxidectin/imidacloprid/ (spot-on) and levamisole (intramuscularly). The different anthelmintic compounds showed variable success regarding their effect on clinical outcome and on stopping egg excretion. Reinfections due to a contaminated environment could not be fully excluded. In winter 2016 and 2017, 218 foxes from the canton of Zurich, Switzerland, were examined. Tissues of nasal and paranasal sinuses were investigated for adult Capillaria specimens and eggs. We describe for the first time C. boehmi infections in Switzerland, observing a high prevalence (190/218, 87.2 %). Overall, 107 of 126 adults (84.9 %, 95 % Confidence Interval, CI: 77.5–90.7 %) and 83 of 92 youngsters (90.2 %, CI: 82.2–95.4 %) were infected. The presence of C. boehmi did not correlate with age (P = 0.209), but correlated significantly with sex: male foxes (102 of 107, 95.3 %, CI: 89.4–98.5 %) were significantly (P = 0.001) more often infected than females (88 of 111, 79.3 %, CI: 70.5–86.4 %). Worm burden ranged from 1 to 72 adult specimens (geometric mean: 5.7). In conclusion, C. boehmi infections can be mis- and/or underdiagnosed in dogs. Appropriate anthelmintic treatments, preventing coprophagia and egg contamination of the surroundings and performing coproscopic controls after treatments are fundamental aspects. Potentially, nasal washing may represent an auxiliary alternative. However, the successful elimination of C. boehmi infections in dogs remains challenging.

Estimation of Serum Copper Concentrations in Sheep and Goats Infected with Gastrointestinal Parasites in Mosul City, Iraq

The objective of the present study was to determine serum copper concentrations in sheep and goats infected with different gastrointestinal parasites. A total of 60 sheep and 30 goats were obtained from different areas in Mosul city and examined in the Veterinary Teaching Hospital, University of Mosul. Fecal and blood samples were collected from each animal. Animals infected with gastrointestinal parasites were diagnosed by detecting parasite eggs in fecal samples. Copper concentrations and hematological values were compared between infected and non-infected animals. Different gastrointestinal parasites were diagnosed include; Chabertia ovina, Cooperia spp., Haemonchus contortus, Marshallagia marshalli, Nematodirus spp., Oesophagostomum spp., Ostertagia ostertagia, Strongylus spp. and Trichostrongylus spp. in both single and mixed infection types. Infected sheep and goats suffered from different clinical signs such as emaciation, poor wool or hair, trembling, pale mucous membrane, loss of appetite, and diarrhea. Infected animals showed significantly (p≤0.05) lower serum copper concentrations compared to non-infected animals. Moreover, hematological parameters (RBC, Hb, PCV, MCH, MCV, and MCHC) were lower compared to non-infected animals. In conclusion, gastrointestinal parasitic infection in sheep and goats can decrease serum copper concentrations and blood parameters in infected animals, thereby exacerbating clinical signs.

Prevalence, Species Distribution, and Related Factors of Fish-Borne Trematode Infection in Ninh Binh Province, Vietnam.

Background. Clonorchis sinensis/Opisthorchis viverrini and minute intestinal flukes (MIF) such as Haplorchis pumilio and H. taichui are fish-borne trematodes (FBT) that may coexist in regions where local people have a habit of eating raw fish like Vietnam. Responses to FBT should be verified according to the data on the distribution of these flukes. This study aims to explore the prevalence of different species of FBT and related factors among local people in a northern province of Vietnam. Methods. A cross-sectional study was conducted in Kim Son and Yen Khanh districts, Ninh Binh province, between March 2016 and March 2017. Four hundred people aged 15 years or older were interviewed and gave stool samples. The FBT eggs in faecal samples were enumerated by modified formalin-ether technique and identified by sequencing of the second internal transcribed spacer (ITS2) region. Result. Among the 400 persons, 19.5% were infected with FBT. On univariate analysis, eating raw fish was the main risk factor (odds ratios (OR)) of 6.769 (95% confidence interval (CI) of 2.655–17.259) followed by being of male gender (3.994 (CI95% 2.117–7.536)) and drinking alcohol (2.680 (CI95% 1.440–4.986)), respectively. There was no risk of increased infection among those living at home without hygienic latrines, those living close to rivers or having ponds, or those raising cats or dogs. By multivariate analysis, FBT infection was only related to the consumption of raw fish and gender. Seventy stool samples with a sufficient amount of faecal matter were subjected to DNA extraction, 42.85% of them yielded DNA production, and all were of Clonorchis sinensis. Conclusion. Results of the study showed the high prevalence of infection of fish-borne trematode, mostly C. sinensis among humans in Ninh Binh province. The prevention of FBT should be strengthened with programs detailed according to the distribution of FBT in different endemic areas.

Comparative Analysis of Haematological Parameters in Hookworm and Plasmodium falciparum Co-Infected Individuals in Kintampo North Municipality, Ghana

Background: Hookworm and Plasmodium falciparum are endemic in Ghana, especially in the north-east and middle belt areas. Haematological parameters have been shown to predict the presence of these infections in patients. This study investigated the effect of hookworm-P. falciparum co-infection on haematological parameters among the patients.&#x0D; Methodology: Stool and blood samples from 984 participants in a cross-sectional study conducted in the Kintampo North Municipality of Ghana were examined for hookworm and Plasmodium falciparum parasites. Malaria parasitaemia was estimated by microscopy and Plasmodium falciparum-specific 18S rRNA gene by polymerase chain reaction. Hookworm eggs in faecal samples were analyzed using Kato-Katz and formol-ether concentration methods. Hookworm speciation was done by PCR. Estimation of haematological parameters was done by automated haematology analyzer. Tukey multiple comparisons test was used to compare continuous variables among the infected groups and Spearman’s rank correlation test determined the relationships between variables.&#x0D; Results: The overall mean (SD) age of the study participants was 22.8 (17.4). Mean lymphocytes and eosinophils counts were higher while mean neutrophil and monocytes counts were lower in the co-infected individuals compared to the single infections. The mean red blood cell (RBC) count and haemoglobin (Hb) levels were higher in the co-infected individuals than in those with malaria only infection. However, white blood cell count and platelet counts were not statistically significant among the groups. There was a significant negative correlation between malaria parasite density and Hb level (r = - 0.239, p = 0.001)&#x0D; Conclusion: Hookworm and Plasmodium falciparum co-infections showed an increased in lymphocyte, eosinophil count and Haemoglobin levels, but a decrease in neutrophil and monocyte levels compared to malaria only infection. The possible mechanisms accounting for the variations in haematological levels remain to be elucidated.

Egg autofluorescence and options for detecting peanut agglutinin binding for the identification of Haemonchus contortus eggs in fecal samples

Quantifying eggs from Haemonchus and other trichostrongyle genera in sheep and goat fecal samples is important for evaluating control and treatment strategies for this family of nematodes with divergent pathologies, capabilities for anthelmintic resistance and environmental susceptibilities. Unfortunately, egg morphology among most of the genera do not differ enough to support the accurate identification of these genera with standard microscopic techniques. Several studies have identified specific lectins which bind selectively to sugars located on the egg surfaces for individual genera among the trichostrongyles. To detect lectins binding to these eggs, they must be directly or indirectly bound to fluorophores, and observed with an epi-fluorescence microscope. The binding of multiple lectins to isolated eggs from a fecal sample can be simultaneously detected if fluorophores are used whose excitation and emission spectra do not overlap, and this would enable the development of a fluorescence-based diagnostic test that identifies multiple trichostrongyle genera within each sample. The present study compared the usefulness of different, commercially available detection systems for use in detecting lectin binding to trichostrongyle eggs. Comparisons were made using the detection of PNA binding to H. contortus eggs with the goal of finding three systems with color spectra that do not overlap. These evaluations included both fluorophores directly conjugated to PNA in a one-step incubation protocol and a two-step incubation protocol involving biotinylated PNA and streptavidin conjugated to different fluorophores. Autofluorescence can affect the efficiency of any fluorescence-based detection system, and significant autofluorescence was observed among the unstained H. contortus eggs with the DAPI-type fluorescence filter, but it was significantly lower with the FITC-type filter and was virtually absent with the rhodamine-type filter. This study demonstrated that all the PNA detection methods tested with H. contortus eggs generated fluorescence intensities (FIs) that were significantly above the autofluorescence generated by the eggs among the three different fluorescence filters. Fluorescence intensities from PNA directly conjugated to either the FITC or rhodamine fluorophores were not different, but the lower autofluoresence in the rhodamine-type filter will enable this fluorophore to be detected more efficiently. Use of biotinylated PNA combined with streptavidin-conjugated to synthetic fluorophores (Alexa Fluor 405, 488 and 546) significantly increased FIs over that of the directly conjugated PNA, but there were no significant differences in FIs among these three biotin-avidin conjugation fluorophores. This biotin-avidin system required two incubation steps. Doubling the concentration of PNA also provided increased FI, at least for the biotin-avidin system. Adding an additional amplification step to the biotin-avidin system involving biotinylated anti-streptavidin followed by the streptavidin-Alexa Fluor complex also provided additional fluorescence.

Development of a lateral flow recombinase polymerase assay for the diagnosis of Schistosoma mansoni infections

Infection with Schistosoma mansoni causes intestinal schistosomiasis, a major health problem across Africa. The accurate diagnosis of intestinal schistosomiasis is vital to inform surveillance/control programs. Diagnosis mainly relies on microscopic detection of eggs in faecal samples but many factors affect sensitivity. Molecular diagnostics are sensitive and specific but application is limited as necessary infrastructure, financial resources and skilled personnel are often lacking in endemic settings. Recombinase Polymerase Amplification (RPA) is an isothermal DNA amplification/detection technology that is practical in nearly any setting. Here we developed a RPA lateral flow (LF) assay targeting the 28S rDNA region of S. mansoni. The 28S LF-RPA assay's lower limit of detection was 10pg DNA with the lower test parameters permitting sufficient amplification being 6 min and 25°C. Optimal assay parameters were 40–45°C and 10 min with an analytical sensitivity of 102 copies of DNA. Additionally the PCRD3 lateral flow detection cassettes proved more robust and sensitive compared to the Milenia HybriDetect strips. This 28S LF-RPA assay produces quick reproducible results that are easy to interpret, require little infrastructure and is a promising PON test for the field molecular diagnosis of intestinal schistosomiasis.

Comparison of Four Diagnostic Methods for Detection and Relative Quantification of Haemonchus contortus Eggs in Feces Samples.

We compared four methods for identification of Haemonchus contortus eggs. With increased trade in animals within and between countries and continents, it has become important to correctly identify H. contortus eggs in fecal samples. To validate the outcome of diagnostic tests, sheep feces (n = 38) were collected from naturally infected flocks in Sweden. Subsamples were analyzed with (a) McMaster egg counting; (b) differential counting of eggs after staining with peanut agglutinin (PNA); (c) detection of DNA following amplification by real-time quantitative polymerase chain reaction (qPCR); and (d) loop-mediated isothermal amplification (LAMP). Differences between similar tests (microscopic and molecular) and SD (±SD) were analyzed with Bland–Altman plots and Spearman rank correlation. Strongylid egg counts ranged from 200 to 12,100 eggs per gram (epg) (mean epg ± SD = 1,278 ± 2,049). Microscopy showed presence of H. contortus eggs in 27 (73%) unstained samples and in 28 (76%) samples stained with PNA, whereas 29 samples (78%) tested positive in LAMP and 34 (91%) in qPCR analysis. The cycle threshold (Ct) values with LAMP ranged between 13 and 38 (mean ± SD = 21 ± 7), and those in qPCR between 25 and 49 (mean ± SD = 33 ± 6). In the LAMP and qPCR analyses, seven (19%) and three (8%) samples, respectively, had a cycle threshold (Ct) >35, whereas no reactions were observed in eight (22%) and three (8%) samples, respectively. There was good agreement between the diagnostic tests based on microscopic examination and DNA detection, although the molecular tests were more sensitive. The bias between the microscopy methods (−4.2 ± 11) was smaller than for the molecular tests (−9.8 ± 10). The observed ranking in terms of test sensitivity was: McMaster counting by conventional microscopy < PNA < LAMP < qPCR. In conclusion, H. contortus can be identified by McMaster counting, without major mistakes regarding false positive results. However, molecular methods provide the capacity to diagnose H. contortus eggs with increased accuracy. This is essential when animals are investigated in quarantine or in studies evaluating anthelmintic treatment efficacy. These methods could also be applied to fecal samples from wildlife to investigate nematode transmission between wildlife and livestock.

PREVALENCE OF HELMINTH EGGS IN CAT FECES CONTAMINATING PUBLIC AREAS IN SURABAYA

Helminthiasis can be transmitted from animals to humans (zoonosis). Helminthiasis can cause cutaneus larva migrants, visceral larva migrant, and occular larva migrants. Cats are the most easily animals can found in public areas. cats have a habit of defecating in areas, such as dusty soil, gardens, sand pits, trash cans, and even children’s playgrounds. Proximity of human life with a stray cats is one of the potential that can helminthiasis transmited to humans. The purpose of this study was to assess the prevalence of helminth eggs (species and number) observed in cat feces contaminating public areas in Surabaya. Cross-sectional study have been observations cats existense and examination laboratory of 180 cat fecal samples were collected from canteens, markets, villages, schools, and parks across 5 areas in Surabaya. Helminth eggs present in fecal samples were identified using direct smear, sedimentation, and flotation methodes, and quantified as fecal egg count (eggs per gram of feces) with McMasster method. The test results positive for helminthiasis if found one or more types of helminth eggs in fecal samples. Helminth eggs were present in 68 (37.8%) of the 180 cat fecal samples contaminating public areas in Surabaya. Results of chi-squared analysis confirmed the prevalence of helminth eggs in cat fecal samples contaminating canteen, markets, villages, schools, and parks in Surabaya (p &gt; 0.05). The species causing environmental contamination included Ancylostoma sp. eggs, Toxocara cati eggs, and Toxascaris leonina eggs. The level of environmental contamination, as assessed using ANOVA, was 200 eggs per gram of feces.