RESUMEN
BACKGROUND: The zoonotic hookworms Ancylostoma caninum and Uncinaria stenocephala are widespread soil-transmitted helminths in dogs in Europe. Given the veterinary and public health importance of hookworms in dogs and the recent changes in the molecular epidemiology of some species, there is a need to continuously monitor the epidemiological and molecular prevalence of these parasites also at the "local" level. The present study aimed to update the epidemiological scenario of hookworm infections in both owned and stray dogs in southern Italy and to discriminate between different hookworm species (A. caninum and U. stenocephala) through molecular analyses. For this purpose, a retrospective analysis was performed over 10 years (2011-2021), including a total of 7008 owned dogs and 5642 stray dogs referred to our laboratory for copromicroscopic examinations. Moreover, 72 faecal samples, from dogs naturally infected by hookworms, were used to discriminate between A. caninum and U. stenocephala using two PCR protocols. Prior to molecular analyses, a subsample of 40/72 positive faecal samples was used for morphometric investigations on hookworm eggs. RESULTS: The results of the ten-year retrospective analysis (2011-2021) showed an overall prevalence of hookworm infection of 9.16%, specifically 5.1% in owned dogs and 14.2% in stray dogs. Logistic regression showed a significant association between positivity to hookworms and the variable "puppies" both in stray (13.84%; OR = 2.4) and owned (7.07%; OR = 2.2) dogs. The results of molecular analyses showed that positivity was confirmed only in 21/72 samples, specifically, 6 samples using protocol A and 19 with protocol B. Sequencing revealed 15 samples positive to U. stenocephala and 6 to A. caninum. CONCLUSIONS: The findings of this study showed a high prevalence of hookworm infections in dogs in southern Italy, updating the epidemiological scenario of the last decade. Moreover, the results of the study revealed the first identification of hookworm species in dogs in Italy by molecular studies, highlighting that U. stenocephala is more prevalent than A. caninum.
Asunto(s)
Enfermedades de los Perros , Infecciones por Uncinaria , Animales , Perros , Ancylostomatoidea/genética , Estudios Retrospectivos , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/parasitología , Infecciones por Uncinaria/epidemiología , Infecciones por Uncinaria/veterinaria , Infecciones por Uncinaria/parasitología , Italia/epidemiología , Heces/parasitología , Ancylostoma/genéticaRESUMEN
The aim of this study was to characterize the Tunisian Fasciola spp. flukes by morphometric and molecular analyses. Flukes were collected from livers of sheep slaughtered in Sejnane slaughterhouses (Bizerte gouvernorate, Northwest Tunisia) between January and March 2021.Five morphometric parameters were determined for all the liver flukes, as follows: (i) total body length (BL), (ii) distance between ventral sucker and the tail (VS-T), (iii) distance between oral sucker and ventral sucker (OS-VS), (iv) abdomen diameter (AD), (v) tail diameter (TD) and the body length to width ratio (BL/BW). Molecular identification of the fluke specimens was carried out by polymerase chain reaction, restriction fragment polymorphism (PCR-RFLP) of a 680 bp sequence of the internal transcribes spacer 1 (ITS1) gene and by amplification, sequencing, and phylogenetic analysis of a 500 bp sequence of the ITS2 gene. Morphometric measurements showed that the mean of the total body length of the adult flukes was 21.1 ± 2.7 mm with minimum and maximum lengths of 13 and 31 mm, respectively. The PCR-RFLP analysis revealed a single profile consisting of three bands of approximately 370, 100, and 60 bp. Fasciola sequences described in the present study (GenBank numbers: OQ457027 and OQ457028) showed 99.58-100% identity to Fasciola hepatica. In conclusion, the results of this study show that molecular and phylogenetic analyses confirm the presence of a single species of F. hepatica in the Sejnane region Northwest of Tunisia. However, further studies are needed to identify the occurrence of Fasciola species in other Tunisian regions.
Asunto(s)
Enfermedades de los Bovinos , Fasciola hepatica , Fasciola , Fascioliasis , Ovinos/genética , Animales , Bovinos , Fasciola/genética , Filogenia , Túnez/epidemiología , Fascioliasis/epidemiología , Fascioliasis/veterinaria , Fasciola hepatica/genética , Enfermedades de los Bovinos/epidemiología , ADN de Helmintos/genéticaRESUMEN
Toxoplasmosis is a zoonotic disease caused by the protozoan parasite Toxoplasma gondii. Infection in humans has usually been related to the consumption of raw, undercooked or cured meat. The aim of this study was to develop a droplet digital polymerase chain reaction (ddPCR)-based assay for the detection and quantification of T. gondii in meat samples. To optimize the ddPCR, T.gondii reference DNA aliquots at five known concentrations: 8000 cg/µl, 800 cg/µl, 80 cg/µl, 8 cg/µl were used. Moreover, results obtained by ddPCR and quantitative PCR (qPCR) were compared using 80 known samples (40 positive and 40 negative), as well as 171 unknown diaphragm tissue samples collected at slaughterhouses. The ddPCR showed a sensitivity of 97.5% and a specificity of 100%, with a detection limit of 8 genomic copy/µl of T. gondii. A nearly perfect agreement (κ = 0.85) was found between results obtained by ddPCR and qPCR for both positive and negative known samples analysed. On the 171 diaphragm tissue samples from field, 7.6% resulted positive by ddPCR and only 1.2% by qPCR. Therefore, this innovative method could be very useful for the detection of T. gondii in meat samples, aiming to prevent human infections.
Asunto(s)
Toxoplasma , Toxoplasmosis Animal , Toxoplasmosis , Animales , ADN Protozoario/genética , Humanos , Carne/parasitología , Reacción en Cadena en Tiempo Real de la Polimerasa , Toxoplasma/genética , Toxoplasmosis/diagnóstico , Toxoplasmosis Animal/diagnóstico , Toxoplasmosis Animal/parasitologíaRESUMEN
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.
Asunto(s)
Enfermedades de los Bovinos/diagnóstico , Heces/parasitología , Microscopía/instrumentación , Microscopía/métodos , Infecciones por Nematodos/veterinaria , Recuento de Huevos de Parásitos/instrumentación , Animales , Bovinos , Enfermedades de los Bovinos/parasitología , Imagenología Tridimensional/veterinaria , Infecciones por Nematodos/diagnóstico , Infecciones por Nematodos/parasitología , Estadísticas no ParamétricasRESUMEN
BACKGROUND: Ticks and tick-borne diseases are increasingly recognized as a cause of disease in dogs worldwide. The epidemiology of ticks and tick-transmitted protozoa and bacteria has changed due to the spread of ticks to urban and peri-urban areas and the movement of infected animals, posing new risks for animals and humans. This countrywide study reports information on distribution and prevalence of pathogens in ticks collected from privately-owned dogs in Italy. We analyzed 2681 Ixodidae ticks, collected from 1454 pet dogs from Italy. Specific PCR protocols were used to detect i) Piroplasms of the genera Babesia and Theileria, ii) Gram-negative cocci of the family Anaplasmataceae and iii) Borrelia burgdorferi sensu lato. Sequencing of positive amplicons allowed for species identification. RESULTS: Babesia/Theileria spp. DNA was detected in 435 homogeneous tick-pools (Minimum Infection Rate (MIR) = 27.6%; 95% confidence interval (CI) = 25.4-29.8%) with higher prevalence in Ixodes ricinus and Rhipicephalus sanguneus group. The zoonotic B. venatorum was the most prevalent species (MIR = 7.5%; 95% CI = 6.3-9.0%). Anaplasma and Ehrlichia species were detected in 165 tick-pools (MIR = 10.5%; 95% CI = 9.3-11.8%) and specifically, A. phagocytophilum was identified with MIR = 5.1% (95% CI = 4.1-6.3%). Borrelia burgdorferi s.l. and B. afzelii were detected with MIR = 0.4% (95% CI = 0.2-0.8%) and MIR = 0.3% (95% CI 0.1-0.7%) respectively. CONCLUSIONS: Zoonotic pathogens B. venatorum and A. phagocytophilum were the most frequently detected in ticks collected from privately-owned dogs which might be used as markers of pathogens presence and distribution.
Asunto(s)
Enfermedades de los Perros/epidemiología , Ixodidae/microbiología , Ixodidae/parasitología , Enfermedades por Picaduras de Garrapatas/epidemiología , Anaplasma/aislamiento & purificación , Animales , Vectores Artrópodos , Babesia/aislamiento & purificación , Borrelia , ADN Bacteriano , ADN Protozoario , Enfermedades de los Perros/microbiología , Enfermedades de los Perros/parasitología , Perros , Ehrlichia/aislamiento & purificación , Femenino , Italia/epidemiología , Masculino , Reacción en Cadena de la Polimerasa , Theileria/aislamiento & purificación , Infestaciones por Garrapatas/veterinaria , Enfermedades por Picaduras de Garrapatas/microbiología , Enfermedades por Picaduras de Garrapatas/parasitologíaRESUMEN
Sarcocystosis is a protozoal disease affecting a wide range of animals. The aims of this study were to characterize the following in sheep: (1) the muscle pathology in Sarcocystis infection, (2) the inflammatory infiltrate and its relationship to severity of infection, and (3) immune markers expressed by parasitized muscle fibers and parasitic cysts. Skeletal muscle samples from 78 sheep slaughtered in southern Italy were snap frozen and analyzed by histopathology, immunohistochemistry, and immunofluorescence. Polymerase chain reaction (PCR) and sequencing were used for Sarcocystis species identification. All 40 muscle samples tested were PCR-positive for Sarcocystis tenella. Histologically, cysts were identified in 76/78 cases (97%), associated with an endomysial infiltrate of lymphocytes and plasma cells. The T cells were predominantly CD8+, with fewer CD4+ or CD79α+ cells. Eosinophils were absent. Notably, sarcolemmal immunopositivity for major histocompatibility complex (MHC) I and II was found in 76/78 cases (97%) and 75/78 cases (96%), respectively, both in samples with and in those without evident inflammatory infiltrate. The number of cysts was positively correlated with inflammation. In addition, MHC I was detected in 55/78 cyst walls (72%), and occasionally co-localized with the membrane-associated protein dystrophin. The findings suggest that muscle fibers respond to the presence of cysts by expression of MHC I and II. The possible role of MHC I and II in the inflammatory response and on the cyst wall is also discussed.
Asunto(s)
Inflamación/veterinaria , Miositis/veterinaria , Sarcocystis/clasificación , Sarcocistosis/veterinaria , Enfermedades de las Ovejas/patología , Animales , Técnica del Anticuerpo Fluorescente/veterinaria , Inmunohistoquímica/veterinaria , Inflamación/parasitología , Inflamación/patología , Complejo Mayor de Histocompatibilidad/inmunología , Músculo Esquelético/parasitología , Músculo Esquelético/patología , Miositis/parasitología , Miositis/patología , Sarcocystis/genética , Sarcocystis/aislamiento & purificación , Sarcocistosis/parasitología , Sarcocistosis/patología , Ovinos , Enfermedades de las Ovejas/parasitología , Linfocitos T/parasitología , Linfocitos T/patologíaRESUMEN
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.
Asunto(s)
Heces/parasitología , Enfermedades de los Caballos/parasitología , Infecciones por Nematodos/veterinaria , Recuento de Huevos de Parásitos/veterinaria , Enfermedades de las Ovejas/parasitología , Animales , Enfermedades de los Caballos/diagnóstico , Caballos , Nematodos , Infecciones por Nematodos/diagnóstico , Recuento de Huevos de Parásitos/métodos , Reproducibilidad de los Resultados , Ovinos , Enfermedades de las Ovejas/diagnóstico , Manejo de Especímenes/métodos , Manejo de Especímenes/veterinariaRESUMEN
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.
Asunto(s)
Enfermedades de los Perros/diagnóstico , Equinococosis/veterinaria , Echinococcus granulosus/aislamiento & purificación , Animales , Coinfección , Enfermedades de los Perros/parasitología , Perros , Equinococosis/diagnóstico , Equinococosis/parasitología , Echinococcus granulosus/genética , Heces/parasitología , Recuento de Huevos de Parásitos/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Juego de Reactivos para Diagnóstico , Taenia/genética , Taenia/aislamiento & purificaciónRESUMEN
BACKGROUND: The cardiopulmonary nematodes Dirofilaria immitis and Angiostrongylus vasorum are increasingly reported in dogs and are responsible for two diseases with overlapping endemic areas, especially in Europe: dirofilariosis and angiostrongylosis. The reasons for their apparent emergence are unknown, but several factors (e.g. increased disease awareness, better diagnostic tools, climatic changes, seasonal population dynamics and movements of animals) may play a role in the recent rise in reports of infection in the various countries of Europe. The aim of this study was to investigate the seroprevalence of D. immitis (by DiroCHECK® ELISA) and the fecal presence of first stage larvae (L1) of A. vasorum (by FLOTAC) in dogs from 68 kennels of the Campania region (southern Italy). The fecal samples were collected from pooled samples using the box as epidemiological unit. To the authors's knowledge, this is the first cross-sectional survey conducted at regional-scale in Italy and in Europe on the contemporaneous detection of D. immitis antigens and A. vasorum L1 in kennels. RESULTS: Antigens of D. immitis were detected in 24/537 (4.4%; 95% Confidence Interval = 3.0-6.7) dogs in 6 out of the 68 kennels (8.8%; 95% CI = 3.6-18.9). The 24 positive samples for D. immitis antigen were tested also with AngioDetect® and only 1 sample was seropositive for A. vasorum with a prevalence of 4.2%. A. vasorum L1 were detected in dogs from 9 out of the 68 kennels (13.2%; 95% CI = 21.8-44.9). Pooled fecal samples from 25 boxes out of the 1360 analyzed resulted positive to A. vasorum L1 (1.8%; 95% CI = 1.2-2.7). CONCLUSIONS: The present study indicates that cardiopulmonary nematodes are present in Campania region in symptomatic dogs as well as in asymptomatic ones. Therefore, regular parasitological surveillance, appropriate treatment strategies and high quality standard of hygiene are required to guarantee the health and welfare of kennel dogs.
Asunto(s)
Angiostrongylus/aislamiento & purificación , Dirofilaria immitis/aislamiento & purificación , Dirofilariasis/parasitología , Enfermedades de los Perros/parasitología , Infecciones por Strongylida/parasitología , Animales , Antígenos Helmínticos , Coinfección , Dirofilariasis/complicaciones , Dirofilariasis/epidemiología , Perros , Italia/epidemiología , Infecciones por Strongylida/complicaciones , Infecciones por Strongylida/epidemiologíaRESUMEN
Oocysts of Toxoplasma gondii represent one of the most common environmental contaminants causing the zoonotic infection toxoplasmosis. The aim of the present study was to compare the Mini-FLOTAC device with traditional cell counting plates (Kova Slide) for the detection of T. gondii oocysts from feline feces. Two types of experiments were performed: (i) purified oocysts were counted in different dilutions and (ii) specific pathogen free T. gondii-negative cat feces was inoculated with numbers of purified oocysts and counting was performed directly from feces. Our analysis showed a thousand times higher sensitivity of Mini-FLOTAC (5 × 10(2) oocysts) compared to Kova Slide (5 × 10(5) oocysts). Also, when compared by McNemar's test, counting of the purified oocysts showed a higher sensitivity of Mini-FLOTAC compared to Kova Slide, for a dilution of 10(3) oocysts/ml (chi(2) = 6.1; P < 0.05). A better sensitivity was also found with Mini-FLOTAC in dilutions of 10(5) and 10(4) oocysts/ml, when counted from feces (chi(2) = 4.2 and 8.1, respectively, P < 0.05). Our results show that Mini-FLOTAC is more sensitive than traditional methods of T. gondii oocysts detection and quantification is more accurate. Furthermore, Mini-FLOTAC simplicity and cost effectiveness allow it to be used with light microscopes in any laboratory or field conditions. We therefore recommend its use for regular screening. Further studies are needed to validate Mini-FLOTAC for the detection of oocysts in soil and water samples in field conditions.
Asunto(s)
Enfermedades de los Gatos/parasitología , Heces/parasitología , Recuento de Huevos de Parásitos/métodos , Toxoplasma/crecimiento & desarrollo , Toxoplasmosis Animal/parasitología , Animales , Enfermedades de los Gatos/diagnóstico , Gatos , Ratones , Oocistos/citología , Oocistos/crecimiento & desarrollo , Recuento de Huevos de Parásitos/instrumentación , Sensibilidad y Especificidad , Organismos Libres de Patógenos Específicos , Toxoplasma/citología , Toxoplasmosis Animal/diagnósticoRESUMEN
The present study aims to assess the performance of different molecular targets using various matrices of samples for the detection of Uncinaria stenocephala (US) in hookworm infected dogs. To this end, the DNA extraction was performed on the following matrices of samples: (i) larvae of US obtained from experimentally infected dogs with US with different larvae counts per microliter (µl); (ii) pure US eggs suspension in distilled water with different egg counts per µl; (iii) spiked dog fecal samples with different US eggs per gram (EPG) of feces; (iv) feces from dogs naturally infected with hookworm eggs; (v) fecal suspension with hookworm eggs recovered from the FLOTAC apparatus. All the samples were tested with four different PCR protocols targeting specific regions for the detection of both hookworms US and AC as follows: Protocol A (ITS1, 5.8â¯S, ITS2) and Protocol B (18â¯S) for the detection of both species, Protocol C (ITS1) for the detection of AC and Protocol D (ITS1) for the detection of US. The best results were obtained with DNA extracted from US larvae matrix obtained from experimentally infected dogs, showing a detection limit of 3.5 larvae/ml for the protocols A, B and D. A moderate correlation was found between the FLOTAC technique and PCR protocols B and D with respect to fecal samples from dogs naturally infected with hookworms. Indeed, PCR protocols B (18â¯S) and D (ITS1) gave the best results for feces and fecal suspension from naturally infected dogs. However, all the PCR protocols used showed lower sensitivity than FLOTAC technique. Perhaps, isolating US eggs in advance could help to obtain better quality and quantity of DNA, avoiding some notable factors such as inhibitors present in faecal samples. However, a further study is needed to evaluate and standardise a protocol for the recovery of parasitic elements, that could be applied prior to DNA extraction. Therefore, this could lead to a better amplification of US eggs DNA. In conclusion, our results showed that the type of sample (sample-matrix) used for the DNA extraction samples is crucial, as this affects the diagnostic sensitivity of the technique.
Asunto(s)
Ancylostomatoidea , Enfermedades de los Perros , Heces , Infecciones por Uncinaria , Reacción en Cadena de la Polimerasa , Animales , Perros , Enfermedades de los Perros/parasitología , Enfermedades de los Perros/diagnóstico , Heces/parasitología , Ancylostomatoidea/aislamiento & purificación , Ancylostomatoidea/genética , Infecciones por Uncinaria/veterinaria , Infecciones por Uncinaria/diagnóstico , Infecciones por Uncinaria/parasitología , Reacción en Cadena de la Polimerasa/veterinaria , Reacción en Cadena de la Polimerasa/métodos , ADN de Helmintos/aislamiento & purificación , ADN de Helmintos/análisis , Recuento de Huevos de Parásitos/veterinaria , Recuento de Huevos de Parásitos/métodos , Larva , Sensibilidad y EspecificidadRESUMEN
BACKGROUND: Cystic echinococcosis (CE), caused by the larval stage of Echinococcus granulosus sensu lato, is a zoonotic parasitic disease of economic and public health importance worldwide, especially in the Mediterranean area. Canids are the main definitive hosts of the adult cestode contaminating the environment with parasite eggs released with feces. In rural and peri-urban areas, the risk of transmission to livestock as well as humans is high because of the free-roaming behavior of owned/not owned dogs. Collecting data on animal movements and behavior using GPS dataloggers could be a milestone to contain the spread of this parasitosis. Thus, this study aims to develop a comprehensive control strategy, focused on deworming a dog population in a pilot area of southern Italy (Campania region) highly endemic for CE. METHODS: Accordingly, five sheep farms, tested to be positive for CE, were selected. In each sheep farm, all shepherd dogs present were treated every 2 months with praziquantel. Furthermore, 15 GPS dataloggers were applied to sheep and dogs, and their movements were tracked for 1 month; the distances that they traveled and their respective home ranges were determined using minimum convex polygon (MCP) analysis with a convex hull geometry as output. RESULTS: The results showed that the mean daily walking distances traveled by sheep and dogs did not significantly differ. Over 90% of the point locations collected by GPS fell within 1500 mt of the farm, and the longest distances were traveled between 10:00 and 17:00. In all the sheep farms monitored, the area traversed by the animals during their daily activities showed an extension of < 250 hectares. Based on the home range of the animals, the area with the highest risk of access from canids (minimum safe convex polygon) was estimated around the centroid of each farm, and a potential scheme for the delivery of praziquantel-laced baits for the treatment of not owned dogs gravitating around the grazing area was designed. CONCLUSIONS: This study documents the usefulness of geospatial technology in supporting parasite control strategies to reduce disease transmission.
Asunto(s)
Enfermedades de los Perros , Equinococosis , Echinococcus granulosus , Humanos , Adulto , Animales , Perros , Ovinos , Praziquantel/uso terapéutico , Enfermedades de los Perros/tratamiento farmacológico , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/prevención & control , Equinococosis/tratamiento farmacológico , Equinococosis/epidemiología , Equinococosis/prevención & control , ZoonosisRESUMEN
Cystic echinococcosis (CE), caused by the tapeworm Echinococcus granulosus, is a zoonotic parasitic disease that still represents a serious threat to human and animal health worldwide. The Mediterranean basin is recognized as one of the major hotspots of CE due to several factors, including the presence of diverse intermediate host species as well as socio-economic and cultural conditions of local communities. This study aims to take a closer look at epidemiological data on CE in the Mediterranean area and assess the knowledge attitudes and practices of shepherds towards this disease in four countries (Algeria, Greece, Italy and Tunisia), highly endemic for CE, with the final goal of identifying highly endemic risk areas and practices in use which might potentially allow the persistence of E. granulosus infection in these areas. To update the epidemiological scenario of CE in Mediterranean areas, a comprehensive review of peer-reviewed literature on CE prevalence data published during the 2017-2023 period was carried out and, through a geographical information system (GIS), a map displaying the current CE distribution in the Mediterranean area was generated. In addition, a questionnaire survey was conducted through in-depth interviews of the farmers to collect information on their management system as well as knowledge attitudes and practices towards CE. From the farmer-participatory survey some risky practices emerged including the non-regular deworming of dogs or the use of ineffective drugs or dosing, as well as the provision of uncooked animal viscera to dogs. Finally, lower levels of knowledge and awareness of the disease was observed among farmers from North Africa compared with those of European countries. In conclusion, the results obtained highlight that CE is still a very serious problem in Mediterranean areas and increased efforts are needed to promote awareness among farmers and to turn research results into policy in order to reduce the spread of this disease, according to the One Health perspective.
Asunto(s)
Equinococosis , Echinococcus granulosus , Conocimientos, Actitudes y Práctica en Salud , Ganado , Animales , Equinococosis/epidemiología , Equinococosis/veterinaria , Equinococosis/prevención & control , Ganado/parasitología , Perros , Región Mediterránea/epidemiología , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/parasitología , Enfermedades de los Perros/prevención & control , Grecia/epidemiología , Zoonosis/epidemiología , Zoonosis/parasitología , Zoonosis/prevención & control , Humanos , Italia/epidemiología , Prevalencia , Bovinos , Túnez/epidemiología , Argelia/epidemiología , Encuestas y Cuestionarios , Ovinos , Agricultores/estadística & datos numéricosRESUMEN
BACKGROUND: Dogs are considered the main domestic animals that may be a reservoir for Leishmania infantum, the agent of zoonotic visceral leishmaniasis (ZVL) in several countries of the world. The dog may host other Leishmania species, but its epidemiological role in the maintenance and spreading of these parasites is not completely elucidated. Zoonotic cutaneous leishmaniasis (ZCL), caused by Leishmania major, affects thousands of people every year and is particularly diffused in many countries of North Africa and Middle East Asia. In ZCL endemic countries, few reports of L. major-positive dogs have been reported, probably because most human cases occur in poor rural areas where the social role of the dog and its medical management is not well considered. The aim of the present study is to better understand the possible involvement of domestic dogs in the epidemiology of ZCL. METHODS: Our research focused on a well-established endemic focus of ZCL, in the area of Echrarda, Kairouan Governorate, central Tunisia. A total of 51 dogs with no or mild clinical signs of vector borne diseases were selected in small villages where human cases of ZCL are yearly present. All dogs were sampled for the Leishmania spp. diagnosis, by using the following procedures: blood sample for serology and buffy coat quantitative polymerase chain reaction (qPCR), popliteal fine needle aspiration, and cutaneous biopsy punch for lymph node and skin qPCR. RESULTS: The results demonstrated a high percentage (21.6%) of dogs positive at least at one or more test; the most sensitive technique was the lymph node qPCR that detected 8/11 positive dogs. Nine, out of the eleven positive dogs, resulted as infected by Leishmania infantum; ITS1-PCR-sequencing allowed Leishmania major identification in the remaining two cases, both from the popliteal lymph node samples, which can suggest a possible visceral spread of a cutaneous Leishmania species in the dog. Interestingly, one of the two L. major-positive dogs was living in the same house where 6-year-old children showed cutaneous lesions referred to as ZCL. CONCLUSIONS: To our knowledge, this is the first report of L. major-positive dogs in Tunisia, the epidemiological role of which remains under investigation.
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Enfermedades de los Perros , Leishmania major , Leishmaniasis Cutánea , Zoonosis , Perros , Animales , Leishmania major/aislamiento & purificación , Leishmania major/genética , Túnez/epidemiología , Leishmaniasis Cutánea/epidemiología , Leishmaniasis Cutánea/veterinaria , Leishmaniasis Cutánea/parasitología , Leishmaniasis Cutánea/diagnóstico , Enfermedades de los Perros/epidemiología , Enfermedades de los Perros/parasitología , Zoonosis/epidemiología , Zoonosis/parasitología , Zoonosis/transmisión , Humanos , Enfermedades Endémicas/veterinaria , Femenino , Masculino , Reservorios de Enfermedades/parasitología , Reservorios de Enfermedades/veterinariaRESUMEN
A cross-sectional serological survey was conducted to evaluate the prevalence of besnoitiosis in cattle farms located in a region of southern Italy. A geographical information system (GIS) was used in order to uniformly sample the bovine farms (n=88) throughout the entire region. Blood samples were collected from 528 autochthonous cattle and sera were tested for antibodies to Besnoitia besnoiti using an enzyme-linked immunosorbent assay test. The farm prevalence was 83.0% (73/88), and the individual animal prevalence was 44.1% (233/528). The availability of geo-referenced point or areal data on bovine besnoitiosis and the construction of prevalence maps by GIS are suggested for dissemination of information to veterinarians on this emerging infection in cattle.
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Anticuerpos Antiprotozoarios/sangre , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/parasitología , Coccidiosis/veterinaria , Sarcocystidae/inmunología , Animales , Bovinos , Coccidiosis/epidemiología , Coccidiosis/parasitología , Estudios Transversales , Ensayo de Inmunoadsorción Enzimática , Italia/epidemiología , Estudios SeroepidemiológicosRESUMEN
[This corrects the article DOI: 10.3389/fvets.2023.1112036.].
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BACKGROUND: Fasciolosis (Fasciola hepatica) and paramphistomosis (Calicophoron daubneyi) are two important infections of livestock. Calicophoron daubneyi is the predominant Paramphistomidae species in Europe, and its prevalence has increased in the last 10-15 years. In Italy, evidence suggests that the prevalence of F. hepatica in ruminants is low in the southern part, but C. daubneyi has been recently reported at high prevalence in the same area. Given the importance of reliable tools for liver and rumen fluke diagnosis in ruminants, this study evaluated the diagnostic performance of the Mini-FLOTAC (MF), Flukefinder(R) (FF) and sedimentation (SED) techniques to detect and quantify F. hepatica and C. daubneyi eggs using spiked and naturally infected cattle faecal samples. METHODS: Briefly, negative bovine faecal samples were artificially spiked with either F. hepatica or C. daubneyi eggs to achieve different egg count levels: 10, 50 and 100 eggs per gram (EPG) of faeces. Moreover, ten naturally infected cattle farms from southern Italy with either F. hepatica and/or C. daubneyi were selected. For each farm, the samples were analysed individually only with MF technique and as pools using MF, FF and SED techniques. Bayesian latent class analysis (LCA) was used to estimate sensitivity and accuracy of the predicted intensity of infection as well as the infection rate in the naturally infected farms. RESULTS: The outcome of this study showed that the highest number of eggs (F. hepatica and C. daubneyi) recovered was obtained with MF, followed by FF and SED in spiked infected samples at 50 and 100 EPG, while at lower infection levels of 10 EPG, FF gave the best results. Moreover, the sensitivity for all the techniques included in the study was estimated at > 90% at infection levels > 20 EPG for both F. hepatica and C. daubneyi eggs. However, MF was the most accurate of the three techniques evaluated to estimate fluke infection intensity. Nevertheless, all three techniques can potentially estimate infection rate at farm level accurately. CONCLUSIONS: Optimization and standardization of techniques are needed to improve the FEC of fluke eggs.
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Enfermedades de los Bovinos , Fasciola hepatica , Fascioliasis , Paramphistomatidae , Trematodos , Infecciones por Trematodos , Bovinos , Animales , Teorema de Bayes , Enfermedades de los Bovinos/diagnóstico , Enfermedades de los Bovinos/epidemiología , Fascioliasis/diagnóstico , Fascioliasis/veterinaria , Fascioliasis/epidemiología , Infecciones por Trematodos/diagnóstico , Infecciones por Trematodos/veterinaria , Infecciones por Trematodos/epidemiología , Rumiantes , HecesRESUMEN
In southern Italy, the number of autochthonous cases of Dirofilaria immitis in dogs has increased considerably. This also occurs in the Campania region, particularly in coastal areas, where infections with D. immitis and Dirofilaria repens have been reported more frequently. Therefore the aim of the present study was to better investigate the occurrence of Dirofilaria spp. in a local dog shelter in Castel Volturno (Campania region, southern Italy). Briefly, a total of 260 blood samples were analysed for identification of microfilariae (mff) and detection of Dirofilaria immitis antigen. Dogs were classified according to their age (1-3 years; 4-6 years; 7-11 years; > 11 years) and length of stay in the shelter at the time of sampling (dogs that entered in the shelter in the last 4 months; dogs housed in the shelter for more than 4 months up to 2 years; dogs housed for more than 2 years). The modified Knott's test revealed that 195 dogs (75.0%) were positive for circulating mff of Dirofilaria spp. Specifically, 104/260 (40.0%) dogs were positive for D. immitis and 91/260 (35.0%) were positive for D. repens. In addition, 72/260 (27.7%) dogs had both D. immitis and D. repens mff. Antigen testing revealed that 78/260 (30.0%) dogs were positive for D. immitis. However, 26/104 (25.0%) of the dogs with D. immitis mff were antigen-negative. The overall k concordance between the modified Knott's test and the antigenic test was ≤0.2 (poor) (p = 0.000). The results of the logistic regression model showed a significant association between Dirofilaria exposure and the period of time the dogs had spent in the shelter at the time of sampling. Dogs housed in the shelter for 4 months (group 1) and between 4 months and 2 years (group 2) had higher Dirofilaria positivity than dogs in group 3 (housed for more than 2 years) (80.4% vs. 79.6% vs. 62.4%, respectively). Moreover, male dogs and older dogs (between 7 and 11 years of age) were more likely to be infected with Dirofilaria spp.
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Toxoplasmosis, caused by the protozoan Toxoplasma gondii, is one of the main food-, water- and soil-borne zoonotic disease worldwide. Over the past 20 years many papers were published on the transmission of T. gondii by marine animals, including mollusks, which can concentrate the oocysts and release them. Sporulated oocysts may remain viable and infective for 18 months in seawater. Therefore, raw or undercooked bivalve mollusks pose a risk to humans. This study aimed to apply and validate for the first time a very sensitive digital droplet polymerase chain reaction (ddPCR) protocol to detect and quantify T. gondii DNA in mussels. Four concentration levels: 8000 genomic copies (gc)/µL, 800 gc/µL, 80 gc/µL, 8 gc/µL of a T. gondii reference DNA were tested. DNA was extracted from 80 pools of mussels (Mytilus galloprovincialis). Forty pools were contaminated with T. gondii reference DNA and used as positive controls, while 40 pools were used as negative controls. DdPCR reaction was prepared using a protocol, previously developed by the authors, for detection of T. gondii in meat. Amplification was obtained up 8 gc/µL. All infected replicates resulted positive, as well as no droplets were detected in negative controls. The droplets produced in the reaction ranged from 8,828 to 14,075 (average 12,627 droplets). The sensitivity and specificity of ddPCR were 100% (95%CI = 94.3-99.9). In addition, 100 pools of mussels collected in the Gulf of Naples were used to validate the protocol. Of these 16% were positive (95% CI = 9.7-25.0) for T. gondii. Samples were also tested by real-time PCR and no positive samples were found. Data obtained from ddPCR showed good identification of negative and positive samples with higher specificity and efficiency than real-time PCR. This tool could be very useful for a rapid sensitive detection of low DNA concentrations of T. gondii in mussels, reducing the risk of toxoplasmosis in humans.
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Nowadays, the exclusive use of commercial anthelmintics for the treatment of gastrointestinal nematode infections in ruminants is less sustainable due to anthelmintic resistance, as well as the problem of drug residues in animal products and the environment. Therefore, an integrated therapeutic approach is needed, including the search for alternatives to synthetic anthelmintic drugs. The aim of this study was to evaluate the possibility of using the essential oil of peppermint (Mentha x piperita L.) in the control of gastrointestinal nematodes in sheep. For this purpose, the in vitro and in vivo anthelmintic efficacy of this oil and the toxic effects on the hosts were examined. In the in vitro egg hatch test, ovicidal activity varied from 21.0-90.3% depending on the concentration of essential oil used (0.0125, 0.025, 0.049, 0.195, 0.781, 3.125, 12.5, and 50 mg/mL). To some extent, anthelmintic efficacy was confirmed in the in vivo fecal egg count reduction test at a mean dose of 150 mg/kg, with an average reduction of nematode eggs of 26.9 and 46.0% at Days 7 and 14 after treatment, respectively. Furthermore, no toxic effects of applied oil were observed on sheep behavior, kidney, or liver function. The main compounds identified by gas chromatography-mass spectrometry analyzes were menthol (32.6%), menthone (22.0%), menthyl-acetate (10.0%), and isomenthone (9.39%). Due to their complex chemical compositions, numerous bioactive ingredients, and natural origin, herbal formulations represent a potentially valuable alternative for the control of gastrointestinal nematodes in sheep. In this context, the results of the present study showed that peppermint essential oil is one of the promising candidates. Further studies should be performed to collect more data on the safety profile of M. piperita EO in treated animals to find the most appropriate formulation for use in field conditions and to test it against resistant gastrointestinal nematode populations.