Evaluation of Parasight All-in-One system for the automated enumeration of helminth ova in canine and feline feces.

BACKGROUND:  Digital imaging combined with deep-learning-based computational image analysis is a growing area in medical diagnostics, including parasitology, where a number of automated analytical devices have been developed and are available for use in clinical practice. METHODS: The performance of Parasight All-in-One (AIO), a second-generation device, was evaluated by comparing it to a well-accepted research method (mini-FLOTAC) and to another commercially available test (Imagyst). Fifty-nine canine and feline infected fecal specimens were quantitatively analyzed by all three methods. Since some samples were positive for more than one parasite, the dataset consisted of 48 specimens positive for Ancylostoma spp., 13 for Toxocara spp. and 23 for Trichuris spp. RESULTS: The magnitude of Parasight AIO counts correlated well with those of mini-FLOTAC but not with those of Imagyst. Parasight AIO counted approximately 3.5-fold more ova of Ancylostoma spp. and Trichuris spp. and 4.6-fold more ova of Toxocara spp. than the mini-FLOTAC, and counted 27.9-, 17.1- and 10.2-fold more of these same ova than Imagyst, respectively. These differences translated into differences between the test sensitivities at low egg count levels (< 50 eggs/g), with Parasight AIO > mini-FLOTAC > Imagyst. At higher egg counts Parasight AIO and mini-FLOTAC performed with comparable precision (which was significantly higher that than Imagyst), whereas at lower counts (> 30 eggs/g) Parasight was more precise than both mini-FLOTAC and Imagyst, while the latter two methods did not significantly differ from each other. CONCLUSIONS: In general, Parasight AIO analyses were both more precise and sensitive than mini-FLOTAC and Imagyst and quantitatively correlated well with mini-FLOTAC. While Parasight AIO produced lower raw counts in eggs-per-gram than mini-FLOTAC, these could be corrected using the data generated from these correlations.

Prevalence and risk factors of gastrointestinal helminths infection in Brazilian horses: A retrospective study of a 12-year (2008-2019) diagnostic data.

Understanding gastrointestinal parasite distribution is crucial for effective control programs in horses. This study reports the prevalence of helminth infections in horses and selected risk factors (i.e., breed, age, climate, season) by analyzing 19,276 fecal samples from the Laboratory of Veterinary Clinical Parasitology, in Curitiba, Southern Brazil. The analyses were carried out from 2008 to 2019, coming from 153 stud farms located in 60 municipalities of nine Brazilian states. The parasite prevalence was 73.3%, with 72.1% present in the adult population and 80.6% in young horses. Strongyles were present in 100% horse farms. Strongyles had a prevalence of 72.1% with a mean FEC of 453.53 (+/- 717.6). Parascaris spp. had a prevalence of 5.8% and a FEC of 17.11 (+/- 149.2). The tropical wet/monsoon climate (Am) showed the lowest FEC for strongyles and Parascaris spp. when compared to the other climates. In the logistic regression analysis, young horses exhibited 4.6 times higher odds ratio (OR) (3.9-5.5) of Parascaris spp. and 1.2 (1.1-1.4) times higher OR of strongyles egg shedding when compared to adults (P < 0.001). Summer presented a higher risk for Parascaris spp. and Strongyles eggs when compared to the other seasons (P < 0.001). Mangalarga Marchador, Criollo, and Crossbred breeds were identified with higher OR of Parascaris spp. egg shedding than Thoroughbred. The extensive prevalence of strongyles across ages, seasons, breeds, and climates alerts for the risk of clinical manifestations in equines raised on pastures designing optimal health management and parasite control strategies worldwide.

Next-generation sequencing technologies for helminth diagnostics and surveillance in ruminants: shifting diagnostic barriers.

Helminth infections in grazing ruminants are a major issue for livestock farming globally, but are unavoidable in outdoor grazing systems and must be effectively managed to avoid deleterious effects to animal health, and productivity. Next-generation sequencing (NGS) technologies are transforming our understanding of the genetic basis of anthelmintic resistance (AR) and epidemiological studies of ruminant gastrointestinal parasites. They also have the potential to not only help develop and validate molecular diagnostic tests but to be directly used in routine diagnostics integrating species-specific identification and AR into a single test. Here, we review how these developments have opened the pathway for the development of multi-AR and multispecies identification in a single test, with widespread implications for sustainable livestock farming for the future.

Gastrointestinal parasites in captive and free-living wild birds in Goiania Zoo / Parasitas gastrointestinais em aves silvestres em cativeiro e de vida livre no Zoológico de Goiânia

Helminths are endoparasites that infect a variety of bird species. Endoparasite infections can cause severe diseases, including kill captive avian hosts and represents a problem to maintenance and conservation. The eggs and larval forms of these parasites are usually eliminated in the host feces. The main interest of this study is to report the occurrence of eggs and oocysts in feces from captive wild birds in Goiânia Zoo and free-living birds in its surroundings. The fecal samples were subjected to parasitological examination to identify the presence of helminths and to classify their eggs based on morphological characteristics. Eggs of nematode parasites (Positive/N) were identified as Ascaridia spp. in Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) and Rhea americana (2/2); Ascarididae in Pavo cristatus nigripensis (1/1); Capillaria spp. in Ara chloropterus (1/1) and Penelope jacucaca (1/1); Capillaria plagiaticia in Anodorhynchus hyacinthinus (1/2) and Ara spp. (2/2); Capillaria venusta in Rhamphastus tucanus (1/2); and unidentified nematode eggs from Amazona amazonica (2/2). Eimeria spp. oocyst was founded in feces from A. hyacinthinus (1/2). Considering free-living birds, Diphyllobothrium spp. eggs were founded in Ardea alba (2/2) and Nycticorax nycticorax (2/3). Co-infection of Eustrongylides spp. was founded in A. alba (1/2). This is the first occurrence of: Ascaridia spp. parasitizing B. chiriri; and C. venusta parasitizing R. tucanus. In conclusion, the helminth eggs found in the abovementioned host bird species are consistent with those reported in the literature, and treatment and control protocols were based on their identification.

Helmintos são endoparasitas que infectam uma variedade de espécies de aves. As infecções por endoparasitas podem causar doenças graves, podendo matar aves em cativeiro e representam um problema de manutenção e conservação. Os ovos e as formas larvais desses parasitas são geralmente eliminados nas fezes do hospedeiro. O principal objetivo deste estudo é relatar a ocorrência de ovos e oocistos em fezes de aves silvestres em cativeiro no Zoológico de Goiânia e em aves de vida livre em seu entorno. As amostras fecais foram submetidas a exame parasitológico para identificação da presença de helmintos e classificação dos ovos com base nas características morfológicas. Ovos de nematóides (Positivo/N) foram identificados como Ascaridia spp. em Brotogeris chiriri (2/2), Dromaius novaehollandiae (1/1) e Rhea americana (2/2); Ascarididae em Pavo cristatus nigripensis (1/1); Capillaria spp. em Ara chloropterus (1/1) e Penelope jacucaca (1/1); Capillaria plagiaticia em Anodorhynchus hyacinthinus (1/2) e Ara spp. (2/2); Capillaria venusta em Rhamphastus tucanus (1/2); e ovos de nematóide não identificado de Amazona amazonica (2/2). Oocistos de Eimeria spp. foram encontrados nas fezes de A. hyacinthinus (1/2). Considerando pássaros de vida livre, ovos de Diphyllobothrium spp. foram encontrados em Ardea alba (2/2) e Nycticorax nycticorax (2/3). Coinfecção de Eustrongylides spp. foi encontradada em A. alba (1/2). Esta é a primeira ocorrência de: Ascaridia spp. parasitando B. chiriri; e C. venusta parasitando R. tucanus. Em conclusão, os ovos de helmintos encontrados nas espécies de aves hospedeiras acima mencionadas são consistentes com os relatados na literatura, e os protocolos de tratamento e controle foram baseados em sua identificação.

Using worm egg count data to detect and counter trends in equine helminth abundance.

Jan van Dijk, RCVS specialist in veterinary parasitology, describes how data can be used to both increase understanding of trends in equine helminth abundance and drive better treatment of individual horses.

Erratic migration of Ascaridia hermaphrodita (Nematoda: Ascaridida) in a blue-fronted Amazon parrot (Amazona aestiva) from illegal wildlife trade: case report / [Migração errática de Ascaridia hermaphrodita (Nematoda: Ascaridida) em papagaio-verdadeiro (Amazona aestiva) oriundo do comércio ilegal: relato de caso]

Ascaridiasis by Ascaridia hermaphrodita is a common parasitic disease in blue-fronted Amazon parrots (Amazona aestiva) and the contamination occurs by ingestion of eggs. Illegal trade of wild birds can increase the frequency of visceral erratic migration by this parasite. Hence, this report describes the erratic migration of A. hermaphrodita and associated lesions in a blue-fronted Amazon parrot. This is a retrospective study, which originated from several apprehension of illegal birds in the state of Paraíba, Brazil. Necropsy revealed intense intestinal ascarid infestation. Hepatic lesions were also observed associated with the parasite migration. Stereoscopy allowed the identification of the morphology of the parasites. Summarizing, this condition in wild birds is not fully understood and may be associated with illegal trade.(AU)

Ascaridíase por Ascaridia hermaphrodita é uma doença parasitária comum em papagaios-verdadeiros. Sua contaminação ocorre pela ingestão de ovos. O tráfico de aves silvestres pode facilitar a migração errática visceral desse parasita. Este relato descreve a migração errática de A. hermaphrodita e lesões associadas em papagaio-verdadeiro. Trata-se de um relato retrospectivo, analisado a partir de uma série de apreensões de aves no estado da Paraíba, Brasil. A necropsia revelou intensa infestação ascarídea intestinal. Também foram observadas lesões hepáticas em uma ave, associadas à migração do parasita. A estereoscopia permitiu distinguir a morfologia dos parasitas. Resumidamente, essa condição em aves silvestres é pouco conhecida e pode ser considerada, inclusive, associada à comercialização ilegal.(AU)

Precision and spatial variation of cyathostomin mucosal larval counts.

Cyathostomins are pervasive parasites of equids across the world. Larval stages encyst in the mucosa of the cecum, ventral and dorsal colon and can induce an inflammatory response leading to larval cyathostominosis, a life-threatening generalized typhlocolitis. Mucosal digestion is the only gold standard procedure for identifying and quantifying all larval stages. There is a lack of standardization of this technique and several aspects are ambiguous, such as precision of the method and the possibility of spatial variation of mucosal larval counts. The aim of this study was to estimate precision for enumeration of early third stage larvae (EL3) and late third stage/fourth stage (LL3/L4) larvae and investigate spatial variation of encysted counts within large intestinal organs. Six naturally infected and untreated horses aged 2-5 years were euthanized as part of an anthelmintic efficacy study, and the cecum (Cec), ventral colon (VC) and dorsal colon (DC) were collected. Each organ was rinsed, weighed, and visually separated into 3 equally sized sections. Two 5% tissue samples were collected from each section, a total of six replicates per organ. The mucosae were digested, and 2% examined under the microscope for presence of EL3 and LL3/L4 stage larvae. Overall, 59 % of the harvested larvae were EL3s, and 41 % were LL3/L4s. The ventral colons represented 45 % of the total organ weight, and contributed 37 and 41 % of the EL3s and LL3/L4s harvested, respectively. The Cec, representing only 27 % of the weight contributed 23 % of EL3s and 47 % of LL3/L4s. The DC represented 28 % of the total organ weight, and 28 % and 12 % of the total EL3s and LL3/L4s, respectively. Coefficients of variation varied from 33 to 183 % for EL3 counts and 38-245% for LL3/L4 counts. There were no statistically significant associations between EL3 counts and either organ or location. For LL3/L4 counts there were no statistically significant differences between the three locations within organs (p = 0.1166), but the DC had significantly lower counts than the other two organs (p < 0.0001). Increasing the number of mucosal replicates from each organ improved estimation, but required a considerably increased workload. In conclusion, mucosal larval cyathostomin counts are highly variable, complicating their use for treatment efficacy estimation.

Evaluation of the VETSCAN IMAGYST: an in-clinic canine and feline fecal parasite detection system integrated with a deep learning algorithm.

BACKGROUND: Fecal examination is an important component of routine companion animal wellness exams. Sensitivity and specificity of fecal examinations, however, are influenced by sample preparation methodologies and the level of training and experience of personnel who read fecal slides. The VETSCAN IMAGYST system consists of three components: a sample preparation device, a commercially available scanner, and an analysis software. The VETSCAN IMAGYST automated scanner and cloud-based, deep learning algorithm, locates, classifies, and identifies parasite eggs found on fecal microscopic slides. The main study objectives were (i) to qualitatively evaluate the capabilities of the VETSCAN IMAGYST screening system and (ii) to assess and compare the performance of the VETSCAN IMAGYST fecal preparation methods to conventional fecal flotation techniques. METHODS: To assess the capabilities of VETSCAN IMAGYST screening components, fecal slides were prepared by the VETSCAN IMAGYST centrifugal and passive flotation techniques with 100 pre-screened fecal samples collected from dogs and cats and examined by both the algorithm and parasitologists. To determine the diagnostic sensitivity and specificity of the VETSCAN IMAGYST sample preparation techniques, fecal flotation slides were prepared by four different techniques (VETSCAN IMAGYST centrifugal and passive flotations, conventional centrifugal flotation, and passive flotation using OVASSAY® Plus) and examined by parasitologists. Additionally, required sample preparation and scanning times were estimated on a subset of samples to evaluate VETSCAN IMAGYST ease-of-use. RESULTS: The algorithm performance of the VETSCAN IMAGYST closely matched that of the parasitologists, with Pearson's correlation coefficient (r) ranging from 0.83-0.99 across four taxa of parasites, Ancylostoma, Toxocara, Trichuris and Taeniidae. Both VETSCAN IMAGYST centrifugal and passive flotation methods correlated well with conventional preparation methods on all targeted parasites (diagnostic sensitivity of 75.8-100%, specificity of 91.8-100%, qualitative agreement between methods of 93.8-94.5%). Sample preparation, slide scan and image analysis were completed within 10-14 min by VETSCAN IMAGYST centrifugal and passive flotations, respectively. CONCLUSIONS: The VETSCAN IMAGYST scanning system with the VETSCAN IMAGYST sample preparation methods demonstrated a qualitative match in comparison to the results of parasitologists' examinations with conventional fecal flotation techniques. The VETSCAN IMAGYST is an easy-to-use, next generation qualitative and possibly quantitative diagnostic platform that brings expert clinical results into the hands of veterinary clinics.

Accuracy and precision of McMaster and Mini-FLOTAC egg counting techniques using egg-spiked faeces of chickens and two different flotation fluids.

Faecal egg counting techniques (ECTs) are useful tools for assessing anthelmintic efficacy and selecting hosts resistant to parasite infection. McMaster (MM) is one of the most commonly used ECTs, but it suffers from low sensitivity and precision. Mini-FLOTAC (MF) has been proposed to replace MM, but so far has not been evaluated for gastro-intestinal nematode infections in chickens. This study compared sensitivity, precision, and accuracy of MM and MF with two trials using egg-spiked faecal samples ranging from 50-1250 eggs per gram of faeces (EPG). In addition, effects of two flotation fluids with different specific gravities (SG), namely salt (SG = 1.20) and sucrose solutions (SG = 1.32), on accuracy and time-spent for both ECTs were evaluated. Overall sensitivity based on the composite reads across all EPG-levels was 97.1 % for MM and 100 % for MF. MF was, however, more sensitive (P = 0.003) or tended to (P = 0.087) be more sensitive than MM at only the lowest EPG-level (i.e. 50 EPG) using one of the duplicate reads, whereas there was no significant difference at any EPG-level using composite reads. Overall average precision of MF (79.5 %) was higher (P < 0.001) than that of MM (63.4 %) across all EPG-levels. Precision of MM increased from 22 to 87 % with increasing EPG-levels from 50-1250 EPG. Corresponding precision estimates for MF ranged from 76 to 91 %. Overall recovery rate of MM (74.6 %) was significantly higher (P < 0.001) than that of MF (60.1 %). There was no significant difference in recovery rate of spiked-eggs among different EPG-levels (P = 0.833). Recovery rate of MM ranged from 64 % to 79 % across different EPG-levels, while it ranged from 54 % to 64 % with MF without an interaction between ECT and EPG-level (P = 0.701). It took more time (P < 0.001) to process (prepare and read) samples with MF than with MM using the same flotation fluid. The sugar solution tended to (P = 0.100) increase egg-recovery with both ECTs, while increasing (P < 0.001) time-spent for processing the samples. Our data collectively suggest that MM is less sensitive than MF only at around minimum detection level of MM when using unrepeated reads. We conclude that McMaster is faster, relatively more accurate but less precise than Mini-FLOTAC. The sugar solution with higher SG increases accuracy of both techniques at the expense of increased labour time.

Endoparasites of household and shelter cats in the city of Rio de Janeiro, Brazil.

Endoparasitic infections are associated with morbidity in cats. This study aimed to investigate the occurrence of endoparasites among cats of different life stages in the city of Rio de Janeiro, Brazil. The samples were analyzed individually by macroscopic exploration for proglottids and centrifugal-flotation. Stool samples were obtained from household cats (n = 57) and shelter cats (n = 336). Endoparasites were detected in 50.64% of the samples. Among household and shelter cats, 21.05% and 55.66% were infected with endoparasites, respectively. In household cats, the most prevalent endoparasites were Ancylostoma spp. (in 25.0%) and Strongyloides spp. (in 25.0%), followed by Toxocara spp. (in 16.67%), Dipylidium caninum (in 16.67%), Cystoisospora spp. (in 8.33%), and Uncinaria spp. + Ancylostoma spp. (in 8.33%). In shelter cats, the most prevalent endoparasite was Ancylostoma spp. (in 29.41%), followed by Cystoisospora spp. (in 26.20%) and Toxocara spp. (in 16.58%), as well as Cystoisospora spp. + Toxocara spp. (in 8.02%); Ancylostoma spp. + Toxocara spp. (in 11.76%); Cystoisospora spp. + Ancylostoma spp. (in 3.74%); Cystoisospora spp. + Toxocara spp. + Ancylostoma spp. (in 3.21%); and Dipylidium caninum + Ancylostoma spp. (in 0.53%). Endoparasitic infections in cats underscore the need for preventive veterinary care and routine coproparasitologic tests.

Alien parasitic infections in the endangered Ryukyu long-furred rat (Diplothrix legata) on Amami-Oshima Island, Japan.

There is limited knowledge about parasites of the endangered Ryukyu long-furred rat, Diplothrix legata (Murinae, Rodentia) endemic to Okinawa, Tokunoshima, and Amami-Oshima Islands in Japan. In the present study, postmortem histopathological examination of an individual found on Amami-Oshima Island revealed a mixed helminth infection of Calodium hepaticum, Hydatigera taeniaeformis, and Angiostrongylus cantonensis. These helminths are considered non-native to Amami-Oshima Island and are maintained by invasive mammals, such as non-native rats and outdoor cats. This observation presents a new host record for C. hepaticum and H. taeniaeformis and the first record of A. cantonensis in Ryukyu long-furred rat on Amami-Oshima Island.

Endoparasites of household and shelter cats in the city of Rio de Janeiro, Brazil / Endoparasitos de gatos domiciliados e de abrigos da cidade do Rio de Janeiro, Brasil

Abstract Endoparasitic infections are associated with morbidity in cats. This study aimed to investigate the occurrence of endoparasites among cats of different life stages in the city of Rio de Janeiro, Brazil. The samples were analyzed individually by macroscopic exploration for proglottids and centrifugal-flotation. Stool samples were obtained from household cats (n = 57) and shelter cats (n = 336). Endoparasites were detected in 50.64% of the samples. Among household and shelter cats, 21.05% and 55.66% were infected with endoparasites, respectively. In household cats, the most prevalent endoparasites were Ancylostoma spp. (in 25.0%) and Strongyloides spp. (in 25.0%), followed by Toxocara spp. (in 16.67%), Dipylidium caninum (in 16.67%), Cystoisospora spp. (in 8.33%), and Uncinaria spp. + Ancylostoma spp. (in 8.33%). In shelter cats, the most prevalent endoparasite was Ancylostoma spp. (in 29.41%), followed by Cystoisospora spp. (in 26.20%) and Toxocara spp. (in 16.58%), as well as Cystoisospora spp. + Toxocara spp. (in 8.02%); Ancylostoma spp. + Toxocara spp. (in 11.76%); Cystoisospora spp. + Ancylostoma spp. (in 3.74%); Cystoisospora spp. + Toxocara spp. + Ancylostoma spp. (in 3.21%); and Dipylidium caninum + Ancylostoma spp. (in 0.53%). Endoparasitic infections in cats underscore the need for preventive veterinary care and routine coproparasitologic tests.

Resumo Endoparasitoses estão associadas à morbidade em gatos. Este estudo objetivou investigar a ocorrência de endoparasitos em gatos de diversas faixas etárias, na cidade do Rio de Janeiro. As amostras passaram por exploração macroscópica à procura de proglotes de cestódeos e analisadas individualmente por centrífugo-flutuação. Assim, amostras fecais foram obtidas de gatos domiciliados (n = 57) e de abrigos (n = 336). Endoparasitos foram detectados em 50,64% das amostras fecais. Nos gatos domiciliados e de abrigos, 21,05% e 55,66% estavam infectados por endoparasitos, respectivamente. Ancylostoma spp. (25%) e Strongyloides spp. (25%) foram os helmintos mais prevalentes encontrados nas amostras de fezes dos gatos domiciliados, seguido por Toxocara spp. (16,67%), Dypilidium caninum (16,67%), Cystoisospora spp. (8,33%) e a associação de Uncinaria spp. e Ancylostoma spp. (8,33%). Entre os gatos dos abrigos, Ancylostoma spp. estavam presentes em 29,41% das amostras, seguidos por Cystoisospora spp. (26,20%), Toxocara spp. (16,58%) e as associações de parasitos Cystoisospora spp. + Toxocara spp. (8,02%), Ancylostoma spp. + Toxocara spp. (11,76%), Cystoisospora spp. + Ancylostoma spp (3,74%), Cystoisospora spp. + Toxocaraspp. + Ancylostoma spp. (3,21%) e Dipylidium caninum + Ancylostoma spp. (0,53%). Cystoisospora spp. foi o único protozoário encontrado. A presença de endoparasitos reforça a necessidade de cuidados veterinários preventivos e testes coproparasitológicos de rotina para evitar sua disseminação.

Prevalence and Genotyping of Echinococcus Species from Livestock in Kajiado County, Kenya.

Cystic Echinococcosis (CE) is a widespread neglected zoonotic disease and is caused by the larval stage of the dog tapeworm Echinococcus granulosus sensu lato. CE is more frequent in livestock-rearing areas and where people live a nomadic or seminomadic lifestyle such as in Kajiado County, Kenya. There is limited data on CE disease situation in the county of Maasailand; the present study, therefore, reports on the prevalence of CE in cattle, sheep, and goats and their relative importance in CE transmission in Kajiado County. In total, 1,486 livestock (388 cattle, 625 sheep, and 473 goats) slaughtered in two abattoirs were examined for the presence of hydatid cysts in various organs. Cyst isolates were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the NADH dehydrogenase subunit 1 gene (nad1). The overall prevalence of CE was 14.8% (220/1486), while prevalence per livestock species was 15.2% (72/473) in goats, 14.9% (93/625) in sheep, and 14.2% (55/388) in cattle. Out of the 421 cysts isolated, 389 cysts were successfully characterized to be either E. granulosus sensu stricto (s. s.), 356/389 (91.5%), E. canadensis (G6/7), 26/389 (6.7%), or E. ortleppi, 7/389 (1.8%). This record confirms predominance of E. granulosus s. s. in Maasailand and other parts of Kenya, while the importance of E. ortleppi and E. canadensis (G6/7) to the general CE burden in Maasailand might be higher than previously thought. More so, a higher infection pressure for humans by E. granulosus s. s. based on its abundance could be speculated. The study sheds significant light on CE situation in livestock in the nomadic/seminomadic society of the Maasai in Kajiado County and provides good bases to investigate human CE in the area.

MALDI-TOF mass spectrometry as a diagnostic tool in human and veterinary helminthology: a systematic review.

BACKGROUND: Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has become a widely used technique for the rapid and accurate identification of bacteria, mycobacteria and certain fungal pathogens in the clinical microbiology laboratory. Thus far, only few attempts have been made to apply the technique in clinical parasitology, particularly regarding helminth identification. METHODS: We systematically reviewed the scientific literature on studies pertaining to MALDI-TOF MS as a diagnostic technique for helminths (cestodes, nematodes and trematodes) of medical and veterinary importance. Readily available electronic databases (i.e. PubMed/MEDLINE, ScienceDirect, Cochrane Library, Web of Science and Google Scholar) were searched from inception to 10 October 2018, without restriction on year of publication or language. The titles and abstracts of studies were screened for eligibility by two independent reviewers. Relevant articles were read in full and included in the systematic review. RESULTS: A total of 84 peer-reviewed articles were considered for the final analysis. Most papers reported on the application of MALDI-TOF for the study of Caenorhabditis elegans, and the technique was primarily used for identification of specific proteins rather than entire pathogens. Since 2015, a small number of studies documented the successful use of MALDI-TOF MS for species-specific identification of nematodes of human and veterinary importance, such as Trichinella spp. and Dirofilaria spp. However, the quality of available data and the number of examined helminth samples was low. CONCLUSIONS: Data on the use of MALDI-TOF MS for the diagnosis of helminths are scarce, but recent evidence suggests a potential role for a reliable identification of nematodes. Future research should explore the diagnostic accuracy of MALDI-TOF MS for identification of (i) adult helminths, larvae and eggs shed in faecal samples; and (ii) helminth-related proteins that are detectable in serum or body fluids of infected individuals.

Parasite survey on a captive wolf population using classical techniques and ELISA coproantigen detection, USA.

As laws change around the United States, wildlife that were once kept as companion animals are now often confiscated by local authorities. They are then euthanized unless a home is found for them at a sanctuary. Wolf sanctuaries are, therefore, becoming increasingly important for their conservation and management. However, little data is available on best practices for the health management of captive wolves, including data on parasitic diseases. Our objective was to assess the prevalence of parasites of captive wolves combining classical coprological techniques and immunoassays based on the detection of coproantigen of selected canid parasites. Fecal samples of 39 animals were collected upon observation of individual animals defecating. All samples were processed using the Fecal Dx® tests, a suite of coproantigen ELISAs for detection of ascarid, hookworm, whipworm, and Giardia (IDEXX Laboratories Inc.). Out of the 39 samples, 38 were processed using the double-centrifugation sugar flotation (DCSF) and 34 using a modification of the Baermann technique. Twenty-eight samples (71.8%) were positive for hookworm, and none positive for the other parasites tested using coproantigen ELISA. Ancylostoma sp. (26, 68.4%), Eucoleus boehmi (13, 34.2%), and Trichuris sp. (2; 5.3%), and Sarcocystis sp. (13, 34.2%) were detected using DCSF. No metastrongyloid lungworm larvae were found. The Cohen's kappa index (0.97) showed excellent agreement between the hookworm coproantigen ELISA and the DCSF using feces preserved in ethanol for a short period of time. This study provides a baseline on the parasites of captive wolves, and shows that recent innovative diagnostics in veterinary parasitology, developed and optimized for dogs, may be used for assessing the health of wolves.

Use of Mini-FLOTAC and Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals / Utilização do Mini-FLOTAC e Fill-FLOTAC no diagnóstico rápido de infecções parasitárias de mamíferos em zoológicos

Abstract Animals reared in restricted environments are highly susceptible to gastrointestinal infection by helminths and protozoa and therefore zoos are characterized as being parasite-rich environments. Successful implementation of control programs of these parasites in zoo environment depends upon precise and rapid diagnosing of gastrointestinal infections. The aim of this study was to demonstrate the role of the Mini-FLOTAC technique in combination with Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals. Fecal samples were collected from 70 animals in four different zoos located in central and southern Italy. All the samples were analyzed using Mini-FLOTAC in combination with Fill-FLOTAC. Out of the 70 pooled samples examined, 80% (24/30) were positive for at least one parasite. Among the gastrointestinal nematodes, Strongyles were the most frequent (40%), followed by Trichuris spp. (23.3%), Parascaris spp. (13.3%) and Capillaria spp. (3.3%). Among the protozoa, Blastocystis spp., Giardia spp. and Eimeria spp. were detected in 6.6%, 3.3% and 3.3%, respectively. These results show that Mini-FLOTAC in combination with Fill-FLOTAC can be used, not only for rapidly diagnosing parasitic infections in zoo mammals, but also for monitoring control programs in which large numbers of fecal samples need to be examined rapidly and reliably.

Resumo Animais criados em ambiente restritos são altamente suscetíveis a infecção gastrointestinal por helmintos e protozoários, constituindo os zoológicos em ambientes com alta contaminação por parasitos. O sucesso da implementação de programas de controle contra estes parasitos em zoológicos depende do rápido diagnóstico das infecções por parasitas gastrointestinais. O objetivo deste estudo foi demonstrar o papel da técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC no diagnóstico rápido das infecções parasitárias em mamíferos em zoológicos. Amostras de fezes foram coletadas de 70 animais de quatro diferentes zoológicos no centro e sudoeste da Itália. Todas as amostras foram analisadas pela técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC. Do total de 70 pools de fezes examinadas, 80% (24/30) foram positivas para pelo menos um parasito. Entre os nematoides gastrointestinais a maior frequência foi observada para estrongilídeos (40%), seguida por Trichuris spp. (23,3%), Parascaris spp. (13,3%) e Capillaria spp. (3,3%). Entre os protozoários Blastocystis spp., Giardia spp. e Eimeria spp. foram detectados em 6,6%, 3,3% e 3,3%, respectivamente. Estes resultados demonstram que a técnica do Mini-FLOTAC em combinação com o Fill-FLOTAC pode ser utilizada não somente para o diagnóstico rápido das infecções parasitárias em mamíferos em zoológicos, mas também no monitoramento de programas de controle onde grande número de amostras fecais devem ser examinadas de forma rápida e confiável.

Use of Mini-FLOTAC and Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals.

Animals reared in restricted environments are highly susceptible to gastrointestinal infection by helminths and protozoa and therefore zoos are characterized as being parasite-rich environments. Successful implementation of control programs of these parasites in zoo environment depends upon precise and rapid diagnosing of gastrointestinal infections. The aim of this study was to demonstrate the role of the Mini-FLOTAC technique in combination with Fill-FLOTAC for rapidly diagnosing parasitic infections in zoo mammals. Fecal samples were collected from 70 animals in four different zoos located in central and southern Italy. All the samples were analyzed using Mini-FLOTAC in combination with Fill-FLOTAC. Out of the 70 pooled samples examined, 80% (24/30) were positive for at least one parasite. Among the gastrointestinal nematodes, Strongyles were the most frequent (40%), followed by Trichuris spp. (23.3%), Parascaris spp. (13.3%) and Capillaria spp. (3.3%). Among the protozoa, Blastocystis spp., Giardia spp. and Eimeria spp. were detected in 6.6%, 3.3% and 3.3%, respectively. These results show that Mini-FLOTAC in combination with Fill-FLOTAC can be used, not only for rapidly diagnosing parasitic infections in zoo mammals, but also for monitoring control programs in which large numbers of fecal samples need to be examined rapidly and reliably.

Helminth infections and hybridization between Haemonchus contortus and Haemonchus placei in sheep from Santana do Livramento, Brazil / Infecções helmínticas e hibridização entre Haemonchus contortus e Haemonchus placei em ovinos criados em Santana do Livramento, Brasil

Abstract The occurrence and intensity of helminth infections were evaluated in sheep from pastures shared with cattle. In 2015 and 2016, young male sheep acquired in Santana do Livramento, Rio Grande do Sul, Brazil, were finished in integrated crop-livestock system. We selected the 12 sheep that showed the highest number of nematode eggs per gram of faeces to search for worms in the gastrointestinal tract. Haemonchus contortus and Trichostrongylus colubriformis were the major parasites. H. contortus presented mean intensities of 1,159 and 257 worms in 2015 and 2016, respectively. T. colubriformis displayed mean intensities of 4,149 and 2,427 worms in 2015 and 2016, respectively. Of the 127 male specimens of Haemonchus spp. analysed by Polymerase Chain Reaction (PCR), 125 were H. contortus, one Haemonchus placei and one hybrid. Other species detected were Cooperia punctata , Cooperia pectinata, Cooperia spatulata, Cooperia curticei, Ostertagia ostertagi, Teladorsagia circumcincta , Trichostrongylus axei, Nematodirus spathiger , and Trichuris ovis. Twenty lambs presented cysts of Taenia hydatigena in the liver and mesentery. One lamb presented Coenurus cerebralis, the larval stage of Taenia multiceps, in the brain. In conclusion, sheep from pasture shared with cattle presented a high diversity of nematode species. H. contortus and H. placei co-infection occur with consequent hybridization.

Resumo A ocorrência de infecções helmínticas foi avaliada em ovinos que compartilhavam pastagem com bovinos. Em 2015 e em 2016, cordeiros machos foram adquiridos em Santana do Livramento, Rio Grande do Sul, para serem terminados em sistema de lavoura - pecuária (ILP). Em cada ano, 12 cordeiros que tinham maior contagem de ovos nas fezes foram abatidos para recuperação dos vermes. Haemonchus contortus e Trichostrongylus colubriformis foram as principais espécies registradas. H. contortus apresentou intensidade média de 1159 e 257 vermes em 2015 e 2016, respectivamente. T. colubriformis apresentou intensidade média de 4149 e 2427 parasitas em 2015 e 2016, respectivamente. De 127 machos de Haemonchus spp. analisados por "Polymerase Chain Reaction" (PCR), 125 foram identificados como H. contortus, um como Haemonchus placei e um como híbrido. Cooperia punctata, Cooperia pectinata, Cooperia spatulata, Cooperia curticei, Ostertagia ostertagi, Teladorsagia circumcincta , Trichostrongylus axei, Nematodirus spathiger e Trichuris ovis foram as outras espécies de nematódeos registradas. Vinte cordeiros apresentaram cistos de Taenia hydatigena no mesentério e no fígado. Um cordeiro apresentou no cérebro Coenurus cerebralis, o estágio larval de Taenia multiceps . Em conclusão, ovinos criados com bovinos apresentam grande diversidade de nematódeos. A co-infecção de H. contortus e H. placei favorece a produção de híbridos.

Helminth infections and hybridization between Haemonchus contortus and Haemonchus placei in sheep from Santana do Livramento, Brazil.

The occurrence and intensity of helminth infections were evaluated in sheep from pastures shared with cattle. In 2015 and 2016, young male sheep acquired in Santana do Livramento, Rio Grande do Sul, Brazil, were finished in integrated crop-livestock system. We selected the 12 sheep that showed the highest number of nematode eggs per gram of faeces to search for worms in the gastrointestinal tract. Haemonchus contortus and Trichostrongylus colubriformis were the major parasites. H. contortus presented mean intensities of 1,159 and 257 worms in 2015 and 2016, respectively. T. colubriformis displayed mean intensities of 4,149 and 2,427 worms in 2015 and 2016, respectively. Of the 127 male specimens of Haemonchus spp. analysed by Polymerase Chain Reaction (PCR), 125 were H. contortus, one Haemonchus placei and one hybrid. Other species detected were Cooperia punctata , Cooperia pectinata, Cooperia spatulata, Cooperia curticei, Ostertagia ostertagi, Teladorsagia circumcincta , Trichostrongylus axei, Nematodirus spathiger , and Trichuris ovis. Twenty lambs presented cysts of Taenia hydatigena in the liver and mesentery. One lamb presented Coenurus cerebralis, the larval stage of Taenia multiceps, in the brain. In conclusion, sheep from pasture shared with cattle presented a high diversity of nematode species. H. contortus and H. placei co-infection occur with consequent hybridization.

New Perspectives in Equine Intestinal Parasitic Disease: Insights in Monitoring Helminth Infections.

Regular anthelmintic treatment has contributed to anthelmintic resistance in horse helminths. This mass anthelmintic treatment was originally developed owing to a lack of larvicidal drugs against Strongylus vulgaris. The high prevalence of anthelmintic resistance and shortening of strongyle egg reappearance period after avermectins/moxidectins requires epidemiologically appropriate and sustainable measures. Selective anthelmintic treatment is a much-needed deworming approach: More than 50% of adult horses manifest no strongyle egg excretion. In this article, selective anthelmintic treatment procedure is described, with the specific focus on the advantages of an evidence-based, medically appropriate, and sustainable treatment system that slows the development of anthelmintic resistance.