Relative egg extraction efficiencies of manual and automated fecal egg count methods in equines.

The World Association for the Advancement of Veterinary Parasitology recently released new recommendations for the design of fecal egg count (FEC) reduction tests for livestock. These provide suggestions as to the number of animals to be sampled and the minimum number of eggs that must be counted to produce statistically meaningful results. One of the considerations for study design is the multiplication factor of the FEC method to be used; methods with lower multiplication factors require fewer animals to be sampled because they are presumed to count more eggs per test. However, multiplication factor is not the sole determinant of the number of eggs counted by any given method, since different techniques use very different sample extraction methodologies that could affect the number of eggs detected beyond just the amount of feces examined. In this light, we compared three commonly used manual FEC methods (mini-FLOTAC, McMaster and Wisconsin) and two automated methods (Imagyst and Parasight All-in-One) with respect to how many equine strongylid and ascarid eggs they counted in the same samples. McMaster and mini-FLOTAC (multiplication factors of 25x and 5x, respectively) produced the most accurate results of the methods tested but mini-FLOTAC counted approximately 5-times more eggs than McMaster. However, Wisconsin and Parasight (multiplication factor = 1x) counted 3-times more ova than mini-FLOTAC, which was less than the 5-fold difference in their multiplication factors. As a result, these tests perform with multiplication factors more akin to 1.6x relative to mini-FLOTAC. Imagyst, due to its unique sample preparation methodology, does not have a traditional multiplication factor but performed similarly to McMaster with respect to egg recovery.

Epidemiological study on factors influencing the occurrence of helminth eggs in horses in Germany based on sent-in diagnostic samples.

Gastrointestinal nematodes are ubiquitous parasites of grazing equines with Parascaris spp., and strongyles being the most relevant ones regarding the prevalence and potential disease severity. Despite their importance, epidemiological data regarding the presence and egg-shedding intensities of these parasites are scarce. Data from 1067 horse samples collected on German horse farms initially to compare diagnostic methods were used for epidemiological analyses. Due to its higher sensitivity, presence/absence data were based on a combined sedimentation/flotation technique while faecal egg counts were based on Mini-FLOTAC. For strongyles, 46.5% of the samples were positive and the median egg-shedding intensity was 40 (range 5-2590). In multivariate analyses, prevalence and egg-shedding intensity were significantly influenced by season, age group and sample type. The drug used for the last treatment and the number of foals on the yard only affected prevalence while the number of horses on the yard and sex were only significant for egg-shedding intensity. For Parascaris spp., a prevalence of 4.6% and a median egg-shedding intensity of 0 (range 5-905) were observed. In multivariate analyses, the age group, the time since the last anthelmintic treatment, presence and number of foals had significant effects on ascarid prevalence whereas egg-shedding intensity was significantly influenced by age group and season only. Parascaris occurred only on yards with foals, but with an increasing number of foals, Parascaris egg-shedding intensity decreased. Prevalence and egg-shedding intensity were influenced by different but partially overlapping variables for Parascaris and strongyles.

High prevalence of pathogenic Eimeria spp. and the main risk factors associated with infection in goats from a semiarid region of Northeastern Brazil.

This study aimed to analyze the epidemiology of infections caused by Eimeria spp. in goats bred in an extensive production system in Northeast Brazil. To this end, an epidemiological survey was performed, wherein 350 fecal samples were collected from goats ranging from 3 months to 8 years old across 30 farms. The number of oocysts per gram of feces (OPG) was counted using the Mini-FLOTAC method. Species were identified based on their morphological and morphometric characteristics. Frequencies between animal categories were assessed using the chi-square test and known risk factors after multivariate analysis. Eimeria oocysts were found in 99.4% (346/348) of samples and 100% (30/30) of the farms. The OPG was influenced by the age of the animals; therefore, young goats (up to 6 months old) eliminated more oocysts (2302.6 + 5454.8) than adult goats (881.7 + 1551.5). Three pathogenic species were identified, with two demonstrating a high prevalence: Eimeria arloingi (88.83%) and Eimeria christenseni (44.7%), in addition to Eimeria caprina (3.72%). In the study, the habit of ground feeding and the use of cisterns as water sources were identified as risk factors, with odds ratios of 4.04 and 9.82, respectively. However, cleaning facilities at shorter intervals was a protective factor (OR = 0.19). We concluded that there was a high prevalence of Eimeria infection in goats bred in extensive production systems, even those in semiarid regions. This high prevalence can be attributed to the congregation of animals in shaded areas and the substantial accumulation of fecal matter at night.

Current status of intestinal parasitosis among patients attending teaching hospitals in Zagazig district, Northeastern Egypt.

Almost 80% of health problems in the developing world are due to malnutrition and infectious diseases, which are mainly parasitic. Updated records on the prevalence of parasitic infections and the potential risk factors are essential to enhancing control strategies. Therefore, this study was conducted to evaluate the current situation of intestinal parasitism among patients attending teaching hospitals in Zagazig district, Northeastern Egypt. The study involved five hundred cases. They were all subjected to faecal examination using direct smear measure and two commercial faecal concentrators: Mini-Parasep® solvent-free and Mini-FLOTAC procedures. Mini-FLOTAC was performed with two solutions (FS2: saturated sodium chloride and FS7: zinc sulphate). The overall prevalence of intestinal parasitic infections was 56%. Different species were identified, like Giardia lamblia (12.6%), Entamoeba histolytica/dispar (10%), Ascaris lumbricoides (8.8%) and Hymenolepis nana (8.6%). Data analyses revealed a significant association of intestinal parasitism with different socio-demographic features of the participants. Our results showed a better diagnostic performance of Mini-Parasep® in the overall recovery of intestinal parasites. It was more accurate than Mini-FLOTAC in diagnosing both helminths and protozoan infections. Mini-FLOTAC (FS2) exhibited a higher sensitivity than FS7 for helminth recovery (74.6% vs 53.4%), while FS7 was more sensitive for protozoan infections (50.6% vs 43.8%). Intestinal parasitosis remains a challenging health problem in Zagazig city, wherever reliable diagnostic approaches are limited. Thus, our study has proposed the value of the commercial concentrators (Mini-Parasep® and Mini-FLOTAC) as alternative techniques for diagnosing a large variety of parasite species in resource-constrained settings.

The Kubic FLOTAC microscope (KFM): a new compact digital microscope for helminth egg counts.

The Kubic FLOTAC microscope (KFM) is a compact, low-cost, versatile and portable digital microscope designed to analyse fecal specimens prepared with Mini-FLOTAC or FLOTAC, in both field and laboratory settings. In this paper, we present the characteristics of the KFM along with its first validation for fecal egg count (FEC) of gastrointestinal nematodes (GINs) in cattle. For this latter purpose, a study was performed on 30 fecal samples from cattle experimentally infected by GINs to compare the performance of Mini-FLOTAC either using a traditional optical microscope (OM) or the KFM. The results of the comparison showed a substantial agreement (concordance correlation coefficient = 0.999), with a very low discrepancy (−0.425 ± 7.370) between the two microscopes. Moreover, the KFM captured images comparable with the view provided by the traditional OM. Therefore, the combination of sensitive, accurate, precise and standardized FEC techniques, as the Mini-FLOTAC, with a reliable automated system, will permit the real-time observation and quantification of parasitic structures, thanks also to artificial intelligence software, that is under development. For these reasons, the KFM is a promising tool for an accurate and efficient FEC to improve parasite diagnosis and to assist new generations of operators in veterinary and public health.

Ascaris lumbricoides eggs or artefacts? A diagnostic conundrum.

Due to the presence of artefacts in stool samples, the copromicroscopic diagnosis of Ascaris lumbricoides is not always straightforward, particularly in the case of fertilized decorticated eggs. A total of 286 stool samples from 115 schoolchildren in India and 171 adult immigrants in Italy were screened for the presence of A. lumbricoides eggs by both Kato-Katz thick smear and Mini-FLOTAC. If the outer layer of A. lumbricoides eggs was absent, two aliquots of each stool sample were preserved: one for coproculture to identify larvae after development and one to compose a pool of stool for molecular analysis. A total of 64 stool samples (22.4%) were positive for A. lumbricoides using the Kato-Katz thick smear; 36 (56.3%) of these showed mammillated A. lumbricoides eggs, 25 (39.1%) showed elements resembling fertilized decorticated eggs, while three samples (4.7%) showed both mammillated and decorticated eggs. By Mini-FLOTAC, 39 stool samples (13.6%) were positive, while decorticated A. lumbricoides-like eggs were identified as artefacts. These results were confirmed by negative coprocultures and quantitative polymerase chain reaction. Mini-FLOTAC can be used for a reliable diagnosis of A. lumbricoides, thanks to the flotation and translation features which allow a clearer view, resulting in the correct identification of A. lumbricoides eggs.

Comparison among FLOTAC, Kato-Katz and formalin ether concentration techniques for diagnosis of intestinal parasitic infections in school children in an Egyptian rural setting.

The study aimed to compare the diagnostic performance of the Kato-Katz, formalin ether concentration method (FECM) and FLOTAC using Sheather's sugar solution (FS1), saturated sodium chloride (FS2) and zinc sulfate (FS7) for the diagnosis of intestinal parasites among school children, focusing on Schistosoma mansoni. Ninety fecal samples were examined using the above mentioned techniques. The overall infection rate was 87.7%. Concerning protozoa, FLOTAC (FS1 and FS2) and FECM detected nearly equal infection rates (43.3% and 44.4%, respectively) with very good agreement. Kato-Katz diagnosed the highest helminthic infection rate (57.8%) followed by FLOTAC FS7 (44.4%) and FECM showed the lowest helminthic infection rate (27.7%). As for S. mansoni, Kato-Katz showed an infection rate of 38.8% vs FLOTAC (22.2%) and FECM (11.1%). The three techniques detected the same infection rate (11.1%) with egg counts more than 72 eggs/gram of feces. The FLOTAC sensitivity and accuracy for the diagnosis of protozoa were 97% and 99%, respectively. Regarding helminths diagnosis, FLOTAC technique showed higher sensitivity (77%) and accuracy (87%) compared to FECM (48% sensitivity and 70% accuracy). Therefore, FLOTAC can be used synchronously or in replacement to other diagnostic techniques. This can strategically impact future control programmes of intestinal parasitic infections in limited resources settings.

Comparative survey of gastrointestinal parasites in sympatric Iberian ibex (Capra pyrenaica) and domestic goats using molecular host specific identification.

An isolated population of several hundred Iberian ibex (Capra pyrenaica) inhabits the Gerês-Xurés Transboundary Biosphere Reserve (GXTBR) in north-western Iberian Peninsula, in partial sympatry with tens of thousands domestic goats (Capra hircus). This study aimed to assess the prevalence and shedding intensity of gastrointestinal parasites from sympatric herds of domestic goat and Iberian ibex in autumn 2018. A total of 93 pooled faecal samples were collected from GXTBR (39 from domestic goats and 54 from Iberian ibex) and the host species was identified using molecular techniques, whenever defecation was not visualised in the field. Parasitological analysis was achieved by joint Willis flotation/sedimentation, McMaster and Mini-FLOTAC techniques. Seventy-two samples (25 domestic goats and 47 Iberian ibex) were retained for analysis after discarding the least fresh and those with uncertain specific identification. Generalized linear mixed models compared prevalence and shedding intensity between caprine species. Domestic goats showed a non-significant tendency to be more parasitized than Iberian ibex, as assessed by overall prevalence (100.0%, CI95 86.7-100.0 vs. 74.5%, CI95 69.5-84.8), and shedding intensity. This study reveals a similar community of gastrointestinal parasites in an abundant livestock species and an isolated population of wild caprine, living in partial sympatry. It is the first study on the health status of this endangered Iberian ibex population, in close contact with livestock, highlighting the need for further sanitary surveillance.

Preliminary evaluation of a novel, fully automated, Telenostic device for rapid field-diagnosis of cattle parasites.

New ideas for diagnostics in clinical parasitology are needed to overcome some of the difficulties experienced in the widespread adoption of detection methods for gastrointestinal parasites in livestock. Here we provide an initial evaluation of the performance of a newly developed automated device (Telenostic) to identify and quantify parasitic elements in fecal samples. This study compared the Telenostic device with the McMaster and Mini-FLOTAC for counting of strongyle eggs in a fecal sample. Three bovine fecal samples were examined, in triplicate, on each of the three fecal egg-counting devices. In addition, both manual (laboratory technician) and automated analysis (image analysis algorithm) were performed on the Telenostic device to calculate fecal egg counts (FEC). Overall, there were consistent egg counts reported across the three devices and calculation methods. The Telenostic device compared very favourably to the Mini-FLOTAC and McMaster. Only in sample C, a significant difference (P < 0.05) was observed between the egg counts obtained by Mini-FLOTAC and by the other methods. From this limited dataset it can be concluded that the Telenostic-automated test is comparable to currently used benchmark FEC methods, while improving the workflow, test turn-around time and not requiring trained laboratory personnel to operate or interpret the results.

Comparing the Mini-FLOTAC and centrifugal faecal flotation for the detection of coccidia (Eimeria spp.) in kiwi (Apteryx mantelli).

Coccidia (Eimeria spp.) in brown kiwi (Apteryx mantelli) cause significant morbidity and mortality in captive rearing facilities. Monitoring the abundance of this parasite in individual birds is crucial for successful management of kiwi. This research compares the abilities of centrifugal faecal flotations (CFF) and a modified Mini-FLOTAC protocol to detect oocysts. We hypothesised that the Mini-FLOTAC would detect higher oocyst counts. Kiwi dropping samples (n = 10) were homogenized in MgSO4 (SG 1.28) and oocyst counts made with CFFs and Mini-FLOTAC counting chambers, with three replicates for each method. For CFF, 0.5 g of droppings were examined using standard methods. Mini-FLOTAC counts were made using a modified sample preparation compared with the manufacturer's protocol but still used a 1:20 dilution of droppings. Oocysts were quantified using light microscopy at ×100-300 magnification. A linear mixed-effects model by REML showed that oocyst per gram estimates via the Mini-FLOTAC method were 3.2 times higher (95% CI 2.4-4.5, p < 0.01) than the CFF results. This increased detection likely represents a more accurate estimation of parasite shedding and should be considered for use in research or applications requiring more accuracy, cost-effectiveness, or accessibility than the CFF provides.

Gastrointestinal investigation of parasites and Enterobacteriaceae in loggerhead sea turtles from Italian coasts.

BACKGROUND: Caretta caretta is the most abundant sea turtle species in the Mediterranean, and studies on this species have vastly expanded during recent years, including those investigating gut bacterial and parasitic communities. Members of these communities have been reported with variable prevalence and pathogenicity, mainly depending on their host and environment (e.g. lifespan, distribution, habitat, diet, health status and stressors). Indeed, many species commonly inhabiting the sea turtle gastrointestinal tract exhibit an opportunistic behaviour. This study aimed to provide baseline data on enterobacterial and parasitic composition, through bacteriological culture-based methods and the FLOTAC parasitological technique, in cloacal and faecal samples of 30 live Caretta caretta, examined upon their arrival at the Marine Turtle Research Centre (Portici, Italy). RESULTS: Enterobacteriaceae were isolated in 18/23 cloacal samples (78.3%), with Citrobacter and Morganella as the most common genera, followed by Proteus, Enterobacter, Providencia, and Hafnia. Parasitic elements were detected in 11/30 faecal samples (36.7%), with Enodiotrema, Rhytidodes, and Eimeria as most common genera, followed by Pachypsolus and Cymatocarpus. Additionally, Angiodyctium is reported for the first time in this host. The majority (47.8%) of sea turtles hosted exclusively Enterobacteriaceae, whereas 30.4% hosted both parasites and Enterobacteriaceae; the remaining 21.8% hosted neither of the agents. CONCLUSIONS: Bacteria and parasites evaluated in the present study are common in Mediterranean loggerhead sea turtles, with slight differences between the western and eastern basin. Although naturally present in the gastrointestinal system of free-living sea turtles, their relationship with these hosts might range from mutualism to parasitism. Indeed, members of the gut community might express their pathogenic potential in immune-compromised animals, such as those in rehabilitation facilities. Therefore, it is advisable to include in the standard work-up of rescued sea turtles a screening procedure for such opportunistic agents, in order to better evaluate the animal's health status and achieve timely intervention with appropriate treatment, thus improving rehabilitation. Furthermore, data collected from free-living sea turtles represent a starting point for investigating wild populations. However, further studies are needed to clarify the differences between sea turtle's normal gut microbiome and pathobiome.

The recovery of added nematode eggs from horse and sheep faeces by three methods.

BACKGROUND: Nematode infections in horses are widespread across the world. Increasing levels of anthelmintic resistance, reported worldwide in equine parasites, have led to the creation of programs for the control of nematodes based on faecal egg counts (FEC). To improve nematode egg counting in equine faecal samples and establish whether the matrix of equine faeces or the eggs affect the counts, the analytical sensitivity, accuracy and precision of Mini-FLOTAC (combined with Fill-FLOTAC), McMaster and Cornell-Wisconsin techniques were compared. Known numbers of eggs extracted from equine or ovine faeces were added to egg free ovine and equine faeces to give counts of 10, 50, 200 and 500 eggs per gram (EPG) of faeces. RESULTS: The Cornell-Wisconsin significantly underestimated egg counts and McMaster showed a low analytical sensitivity, revealing 100% of sensitivity only for concentrations greater than 200 EPG. EPG values detected by Mini-FLOTAC did not differ significantly from expected counts at any level of egg density. CONCLUSIONS: Mini-FLOTAC combined to Fill-FLOTAC which provides an accurate method of weighing without need for a balance and filtering out debris, could be used for FEC on the farm as well as in the laboratory.

Bacteria and parasites in Podarcis sicula and P. sicula klemmerii.

BACKGROUND: New epidemiological data on bacterial and parasitic infections in 24 Italian wall lizards, namely Podarcis sicula (mainland population) and P. sicula klemmerii (insular population) in southern Italy were provided. To achieve this goal, samples were collected from individuals belonging to the two populations and analysed by microbiological and parasitological methods. RESULTS: A wide range of bacteria (e.g. Pantoea spp., Citrobacter spp., Morganella spp., Pseudomonas, Enterobacter spp., Staphylococcus spp. and Escherichia coli) and parasites (e.g. Ophionyssus natricis, coccidia, Dicrocoelidae) were detected in both P. sicula and P. sicula klemmerii individuals. Insular population presented similar bacterial and parasitic diversity to its mainland counterpart. Ampicillin was the antimicrobial with the highest resistance rate. CONCLUSION: This study highlighted various bacteria and parasites, some of them potentially zoonotic. Further studies are needed to better understand the epidemiology and transmission routes of these pathogens along with their impact on the welfare and behaviour of Italian wall lizards.

Innovative tools for the diagnosis of Echinococcus granulosus in definitive hosts.

The aim of this study was to develop and validate an innovative protocol for the diagnosis of Echinococcus granulosus and other Taeniidae in dogs. For this purpose, three experiments were performed, using faecal samples from naturally infected dogs. Firstly, the FLOTAC technique was calibrated with five flotation solutions: saturated sodium chloride (specific gravity, s.g. = 1.20), zinc sulphate (s.g. = 1.35), zinc chloride (s.g. = 1.45), Breza (s.g. = 1.30) and modified Breza (s.g. = 1.40). Then, FLOTAC was compared with four techniques of flotation in centrifuge using: zinc sulphate (s.g. = 1.20), Breza (s.g. = 1.30), modified Breza (s.g. = 1.40), and zinc chloride (s.g. = 1.45). Finally, four different protocols of DNA extraction were compared for Taeniidae identification: QIAamp Tissue Kit and QIAamp Stool from eggs; QIAamp Stool and Wizard Magnetic Purification System for Food from faeces. FLOTAC with zinc sulphate was the most efficient method to detect Taeniidae eggs, showing highest mean of eggs per gram (EPG) of faeces. The QIAamp Stool, using eggs concentrated by FLOTAC, was the best method for DNA extraction. The combination of these protocols provided the highest number of positive samples with PCR, i.e., 47/50 (94.0%). The three negative samples showed a low faecal egg count (2 EPG) below the detection limit (4 EPG) of the protocol. From sequencing of the 47 positive samples: 6 samples were identified as E. granulosus sensu stricto (s.s.), 28 as Taenia hydatigena and 6 as T. pisiformis. A co-infection between different genera of Taeniidae was found in 7 samples.

Formalin-ethyl acetate concentration, FLOTAC Pellet and anal swab techniques for the diagnosis of intestinal parasites.

The aim of this study was to compare formalin-ethyl acetate concentration and FLOTAC Pellet techniques for the diagnosis of intestinal parasites in human stool samples. The anal swab method was used for the specific detection of Enterobius vermicularis. The study was performed in children and youth from Puerto Madryn (South Argentina). A total of 174 individuals were examined using the formalin-ethyl acetate concentration technique (FECT), the FLOTAC Pellet technique with saturated sodium chloride and zinc sulphate flotation solutions and anal swabs. The performance of copromicroscopic techniques was evaluated according to sensitivity, negative predictive value and Kappa index. Overall, 39.1% of the individuals were parasitised. The most prevalent species was Blastocystis sp. (19%) followed by E. vermicularis (17.8%), Giardia lamblia (6.3%), Entamoeba coli (5.7%), Hymenolepis nana and Endolimax nana (1.1%). The FECT was the most sensitive technique for Blastocystis sp., G. lamblia and E. coli infections, whereas FLOTAC Pellet techniques were the most sensitive for H. nana diagnosis. Anal swabs detected the highest percentage of E. vermicularis infection. This was the first time that the FLOTAC Pellet technique was used to detect intestinal parasites in humans. The FECT continues to be a reliable method for detecting protozoa and the FLOTAC Pellet technique gains importance in the diagnosis of helminths. Anal swab test remains the method of choice for the detection of E. vermicularis. However, when comparing techniques, key factors as preservation methods, preservation times and flotation solutions should be taken into account.

Angiostrongylus vasorum infection in dogs from a cardiopulmonary dirofilariosis endemic area of Northwestern Italy: a case study and a retrospective data analysis.

BACKGROUND: In Italy, Angiostrongylus vasorum, an emergent parasite, is being diagnosed in dogs from areas considered free of infection so far. As clinical signs are multiple and common to other diseases, its diagnosis can be challenging. In particular, in areas where angiostrongylosis and dirofilariosis overlap, a misleading diagnosis of cardiopulmonary dirofilariosis might occur even on the basis of possible misleading outcomes from diagnostic kits. CASE PRESENTATION: Two Cavalier King Charles spaniel dogs from an Italian breeding in the Northwest were referred to a private veterinary hospital with respiratory signs. A cardiopulmonary dirofilariosis was diagnosed and the dogs treated with ivermectin, but one of them died. At necropsy, pulmonary oedema, enlargement of tracheo-bronchial lymphnodes and of cardiac right side were detected. Within the right ventricle lumen, adults of A. vasorum were found. All dogs from the same kennel were subjected to faecal examination by FLOTAC and Baermann's techniques to detect A. vasorum first stage larvae; blood analysis by Knott's for Dirofilaria immitis microfilariae, and antigenic tests for both A. vasorum (Angio Detect™) and D.immitis (DiroCHEK® Heartworm, Witness®Dirofilaria). The surviving dog with respiratory signs resulted positive for A. vasorum both at serum antigens and larval detection. Its Witness® test was low positive similarly to other four dogs from the same kennel, but false positive results due to cross reactions with A. vasorum were also considered. No dogs were found infected by A. vasorum. Eventually, the investigation was deepened by browsing the pathological database of Veterinary Pathology Laboratories at Veterinary School of Milan University through 1998-2016, where 11 cases of angiostrongylosis were described. Two out of 11 dogs had a mixed infection with Crenosoma vulpis. CONCLUSION: The study demonstrates the need for accurate surveys to acquire proper epidemiological data on A. vasorum infection in Northwestern Italy and for appropriate diagnostic methods. Veterinary clinicians should be warned about the occurrence of this canine parasite and the connected risk of a misleading diagnosis, particularly in areas endemic for cardiopulmonary dirofilariosis.

Survey of Baylisascaris spp. in captive striped skunks (Mephitis mephitis) in some European areas.

Skunks are popular carnivore species kept both in zoological institutions and in households where they are hand raised as exotic pets. These small carnivores are considered the main definitive hosts of the roundworm Baylisascaris columnaris. The purpose of this survey was to investigate the occurrence of Baylisascaris spp. in striped skunks kept as pets or in private zoo collections in some European areas. Copromicroscopic data from two laboratories, one in Italy and one in Germany, were used. A total of 60 animals were selected. Samples came from Germany (n = 30), Italy (n = 23), United Kingdom (n = 5), Austria (n = 1), and the Netherlands (n = 1). Twenty-eight animals were certainly kept as pets in private households in Italy and the UK. Fifteen out of 60 animals (25%) were positive for Baylisascaris spp. Molecular identification of adult parasites was performed in ten of those animals, revealing B. columnaris in all cases. To the authors' knowledge, this is the first survey of Baylisascaris spp. in captive skunks in Europe.

Use of the FLOTAC technique to diagnosing parasites of the urinary tract of dogs.

Among the nematodes that infect the urinary tract of dogs, the Dioctophyma renale and Capillaria plica are those more frequently reported. For a long time, sedimentation was the only method used to detect eggs of these parasites in urine. The aim of this study was to analyze urine samples (n = 54) of dogs, obtained by bladder catheterization or cystocentesis, by using a modified FLOTAC technique. Animals were divided into two groups, i.e., with (n = 20) and without (n = 34) suspicion of urinary disease. The overall positivity herein observed was 3.8 % (2/54), being all animals (10 %; 2/20) from the group with suspicion of urinary disease. In the first positive sample, a single egg of D. renale was detected, whereas in the second sample two trematode-like eggs were observed. This is the first short survey employed to detect eggs of parasites that inhabit the urinary tract of dogs using a modified FLOTAC technique; in addition, for the first time, eggs of D. renale have been detected using this tool.

New insights into diagnosis of Platynosomum fastosum (Trematoda: Dicrocoeliidae) in cats.

Platynosomum fastosum is a hepatic trematode which causes the so-called lizard poisoning in cats. This parasite is reported in tropical and sub-tropical areas infecting the liver, gallbladder, and bile ducts of cats. Despite its clinical importance, the diagnosis of P. fastosum has been poorly investigated so far. In this study, three cases of infection by P. fastosum in cats are reported. The FLOTAC technique was utilized for the first time in the detection of eggs of this parasite. Three cats (two males and one female), being one of them with a history of diarrhea and vomiting, were diagnosed positive for the presence of P. fastosum eggs through the FLOTAC technique. In conclusion, this study provides important and new insights into the diagnosis of P. fastosum in cats, expanding the geographical knowledge of this parasite in Brazil.

Immigrants living in an urban milieu with sanitation in Southern Italy: persistence and transmission of intestinal parasites.

In the current era of globalization, the massive movement of populations to developed countries causes a greater attention to neglected tropical diseases in places where such diseases are considered unusual. The present study was planned to assess the persistence of intestinal parasitosis in immigrants stably living in the urban central area of Naples (Southern Italy) and the spread of infection within households with a lifestyle similar to that of the country of origin. A total of 2150 stool samples were analysed with the FLOTAC dual technique, and 415 subjects (19.3 %) tested positive for pathogenic intestinal parasites. One hundred ninety-six subjects were randomly selected and monitored again after 1 year in order to evaluate the persistence of intestinal parasites in immigrants having access to proper sanitation. No pathogenic parasites were found in these 196 samples. A total of 482 cohabitants of 151 positive subjects were recruited to evaluate the interfamilial spread of the identified parasites. Only in 18 households were there subjects infected with the same parasite. Monitoring of parasites in stool samples of immigrants showed a decrease of almost all pathogenic species over the years. From the analysis of households, it is not possible to assert that there is a familial transmission. Our study provides evidence that the prevalence of parasitic infections in immigrants is likely related to the poor sanitary habits of the country of origin and that acquisition of new sanitary regulations, together with the administration of pharmacological treatment, limits the transmission in the households and in the local population of their destination.