146 results on '"Latrofa MS"'
Search Results
2. Molecular detection of Leishmania spp. in dogs and a cat from Doha, Qatar
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Lima, C, Colella, V, Latrofa, MS, Cardoso, L, Otranto, D, Alho, AM, Lima, C, Colella, V, Latrofa, MS, Cardoso, L, Otranto, D, and Alho, AM
- Abstract
Background Canine and feline leishmanioses are increasingly reported worldwide and represent a threat to both animal and human health. Despite their relevance, data about leishmanioses in companion animals in the east-central part of the Arabian Peninsula are unavailable. Therefore, we investigated the occurrence of Leishmania spp. in dogs and cats from Qatar. Methods From March 2016 to May 2018, 199 pets (120 dogs and 79 cats) living in Doha or its outskirts were included in this study. From each animal a blood sample was collected and tested for Leishmania spp. by quantitative real-time PCR. Results Out of the 199 animals, four (2.0%) were positive for Leishmania spp., including three dogs (2.5%) and one cat (1.3%). Conclusions All positive animals were born in Qatar and had not travelled overseas, suggesting that infection was locally acquired. Considering the occurrence of Leishmania spp. and its potential impact on the health of animals and humans, it is crucial to increase scientific knowledge in order to plan screening and regular prophylaxis against sand fly vectors to reduce the risk of infection.
- Published
- 2019
3. A nationwide survey of Leishmania infantum infection in cats and associated risk factors in Italy
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Harley, D, Iatta, R, Furlanello, T, Colella, V, Tarallo, VD, Latrofa, MS, Brianti, E, Trerotoli, P, Decaro, N, Lorusso, E, Schunack, B, Miro, G, Dantas-Torres, F, Otranto, D, Harley, D, Iatta, R, Furlanello, T, Colella, V, Tarallo, VD, Latrofa, MS, Brianti, E, Trerotoli, P, Decaro, N, Lorusso, E, Schunack, B, Miro, G, Dantas-Torres, F, and Otranto, D
- Abstract
Though scantly investigated, Leishmania infantum infection and clinical cases of leishmaniasis in cats have been recently reported in several countries of the Mediterranean basin, with large variability in prevalence data. A major limitation in the comparability of the data available is attributed to the differences in diagnostic techniques employed and cat populations sampled. The aim of this study was to assess the prevalence of L. infantum infection in owned cats across Italy by serological and molecular tests and the identification of potential risk factors. Blood samples from 2,659 cats from northern (n = 1,543), central (n = 471) and southern (n = 645) Italy were tested for antibodies against L. infantum, by an immunofluorescence antibody test and for the parasites’ DNA, by real-time PCR. Samples were additionally screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) proviral DNAs. An overall cumulative L. infantum prevalence of 3.9% was recorded by serology (3.3%) and/or qPCR (0.8%), with a higher rate (10.5%) in southern Italy. The risk of L. infantum infection in cats was significantly associated to the geographical areas (South vs North and Centre; p<0.0001), age class (from 19 months to 6 years old vs ≤18 months old, p = 0.0003), neutering status (not neutered vs neutered, p = 0.0028) and FIV infection (p = 0.0051).Though the role of cats in the epidemiology of L. infantum is still debated, our findings indicate that cats are exposed to and/or infected by this protozoan, mainly in endemic regions of Italy. Hence, a standardization of procedures for a prompt diagnosis of L. infantum infection in cats and for screening cat population is crucial for a better understanding of the epidemiology of feline leishmaniasis, and of the potential role of cats in the transmission cycle of zoonotic visceral leishmaniasis.
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- 2019
4. Prevention of feline leishmaniosis with an imidacloprid 10%/flumethrin 4.5% polymer matrix collar
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Brianti, E, Falsone, L, Napoli, E, Gaglio, G, Giannetto, S, Pennisi, MG, Priolo, V, Latrofa, MS, Tarallo, VD, Basano, FS, Nazzari, R, Deuster, K, Pollmeier, M, Gulotta, L, Colella, V, Dantas-Torres, F, Capelli, G, Otranto, D, Brianti, E, Falsone, L, Napoli, E, Gaglio, G, Giannetto, S, Pennisi, MG, Priolo, V, Latrofa, MS, Tarallo, VD, Basano, FS, Nazzari, R, Deuster, K, Pollmeier, M, Gulotta, L, Colella, V, Dantas-Torres, F, Capelli, G, and Otranto, D
- Abstract
BACKGROUND: Leishmaniosis caused by Leishmania infantum is one of the most important vector-borne diseases affecting animals and humans worldwide. Dogs are considered main reservoirs of the zoonotic forms, though in the last years the role of cats as reservoirs has been increasingly investigated. Feline leishmaniosis (FeL) occurs in endemic areas and no specific preventive measures have been investigated so far. In this study the efficacy of a 10% imidacloprid/4.5% flumethrin polymer matrix collar, licensed for tick and flea prevention, has been assessed against FeL in a longitudinal study on 204 privately owned cats from the Aeolian islands (Sicily), an area highly endemic for the disease. From March to May 2015 [Study Day 0 (SD 0)], cats negative for FeL were collared (G1, n = 104) or left untreated (G2, n = 100). Diagnosis consisted of serology and qPCR on blood and conjunctival swabs, which were collected at baseline (SD 0) and at the end of the study (SD 360). Interim clinical examinations were performed on SD 210 (when collars were replaced in G1) and SD 270. RESULTS: Of the 159 cats which completed the study, 5 in G1 and 20 in G2 were positive for L. infantum infection, in at least one of the diagnostic tests leading to a yearly crude incidence of 6.3% and 25.0% in G1 and G2, respectively (P = 0.0026). This translates into an efficacy of the collar of 75.0% in preventing feline Leishmania infection. The collar was generally well tolerated with no systemic adverse reactions and few local skin reactions were observed in the application area in four out of 104 treated cats (3.8%). CONCLUSIONS: The 10% imidacloprid/4.5% flumethrin collar significantly reduced the risk of L. infantum infection in cats. To our knowledge, this is the first study in which a preventative strategy against feline Leishmania infection is assessed under natural conditions. These findings close a gap in veterinary medicine, in that they confirm this collar as a tool in reducing the risk of
- Published
- 2017
5. Molecular detection of vector-borne pathogens in dogs and cats from Qatar
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Alho, AM, Lima, C, Latrofa, MS, Colella, V, Ravagnan, S, Capelli, G, de Carvalho, LM, Cardoso, L, Otranto, D, Alho, AM, Lima, C, Latrofa, MS, Colella, V, Ravagnan, S, Capelli, G, de Carvalho, LM, Cardoso, L, and Otranto, D
- Abstract
BACKGROUND: Vector-borne diseases (VBDs) have been increasingly reported in dogs and cats worldwide. However, no data are currently available regarding canine and feline VBDs in Qatar and limited information is available from other Persian Gulf countries. METHODS: Blood samples from 98 client-owned animals (i.e. 64 dogs and 34 cats) living in Doha (Qatar) were collected and the presence of genomic DNA of Anaplasma spp., Babesia spp., Dirofilaria spp., Ehrlichia spp., Hepatozoon spp., Mycoplasma spp. and Rickettsia spp. was assessed by polymerase chain reaction (PCR), real time-PCR (rt-PCR) and sequence analysis. RESULTS: Of the 64 dogs, 12 (18.8%) were infected with at least one pathogen (i.e. 7.8% with Mycoplasma spp., 4.7% with Babesia vogeli, 3.1% with Ehrlichia canis, and 1.6% with Anaplasma platys, Babesia gibsoni and Hepatozoon canis, each). One of the 12 dogs was co-infected with B. vogeli and E. canis. Of the 34 cats, seven (20.6%) animals were infected with at least one pathogen (i.e. 5.9% were positive for Mycoplasma spp., and 2.9% for Babesia felis, B. vogeli, E. canis, "Candidatus Mycoplasma haemominutum" and Mycoplasma haemofelis, each). No dogs or cats were positive for Dirofilaria spp. or Rickettsia spp. CONCLUSIONS: Although the sample sizes of dogs and cats herein analysed was moderately small, data from this study report the occurrence of A. platys, B. vogeli, B. gibsoni, E. canis, H. canis and Mycoplasma spp. in domestic dogs and of B. felis, B. vogeli, "Candidatus M. haemominutum", E. canis and M. haemofelis in domestic cats from Qatar. Further investigations along with prophylactic measures are strongly recommended in order to reduce the risk of dogs and cats acquiring VBDs in Qatar.
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- 2017
6. Dirofilaria immitis in pinnipeds and a new host record
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Alho, AM, Marcelino, I, Colella, V, Flanagan, C, Silva, N, Correia, JJ, Latrofa, MS, Otranto, D, de Carvalho, LM, Alho, AM, Marcelino, I, Colella, V, Flanagan, C, Silva, N, Correia, JJ, Latrofa, MS, Otranto, D, and de Carvalho, LM
- Abstract
BACKGROUND: Dirofilaria immitis is a mosquito-borne pathogen that is spreading worldwide, and the associated infection (i.e. dirofilariosis) is becoming a threat to animals and humans living in endemic areas. Little is known about the occurrence and risk of infection of D. immitis in pinnipeds. Here we report dirofilariosis by D. immitis in several pinniped species kept in captivity in Portugal. METHODS: Animals were housed in an oceanographic park located in Algarve, southern Portugal, a geographical area endemic for canine dirofilariosis. To assess the occurrence of D. immitis, blood was collected from the park's resident pinniped population, which consisted of 16 animals (5 common seals Phoca vitulina, 2 grey seals Halichoerus grypus, 3 California sea lions Zalophus californianus and 6 South African fur seals Arctocephalus pusillus pusillus). Dirofilaria immitis nematodes were detected by real-time PCR and by the presence of circulating antigens. In addition, modified Knott's technique was performed to detect circulating microfilariae. Necropsies and histopathological examination of two animals which died during the study were also conducted. RESULTS: Out of the 16 pinnipeds housed at the park, seven (43.8%) were positive for D. immitis by real-time PCR (3 P. vitulina, 2 Z. californianus and 2 A. p. pusillus), two of which (P. vitulina) were also positive for the nematode's antigen. Additionally, D. immitis microfilariae were detected in one A. p. pusillus. Furthermore, several D. immitis specimens were retrieved from the right ventricle and pulmonary arteries at the necropsy of one P. vitulina and one A. p. pusillus. CONCLUSIONS: This study provides new epidemiological data on D. immitis infection in pinnipeds diagnosed through clinical, molecular and pathological findings. Additionally, the South African fur seal is herein reported as a new host for this zoonotic filarioid. The situation herein described could also occur in other parks located in areas where ca
- Published
- 2017
7. Helminth infections and gut microbiota - a feline perspective
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Duarte, AM, Jenkins, TP, Latrofa, MS, Giannelli, A, Papadopoulos, E, de Carvalho, LM, Nolan, MJ, Otranto, D, Cantacessi, C, Duarte, AM, Jenkins, TP, Latrofa, MS, Giannelli, A, Papadopoulos, E, de Carvalho, LM, Nolan, MJ, Otranto, D, and Cantacessi, C
- Abstract
BACKGROUND: Investigations of the relationships between the gut microbiota and gastrointestinal parasitic nematodes are attracting growing interest by the scientific community, driven by the need to better understand the contribution of parasite-associated changes in the composition of the gut flora to both host malnutrition and immune modulation. These studies have however been carried out mainly in humans and experimental animals, while knowledge of the make-up of the gut commensal flora in presence or absence of infection by parasitic nematodes in domestic animals is limited. In this study, we investigate the qualitative and quantitative impact that infections by a widespread parasite of cats (i.e. Toxocara cati) exert on the gut microbiota of feline hosts. METHODS: The faecal microbiota of cats with patent infection by T. cati (= Tc+), as well as that of negative controls (= Tc-) was examined via high-throughput sequencing of the V3-V4 hypervariable region of the bacterial 16S rRNA gene, followed by bioinformatics and biostatistical analyses of sequence data. RESULTS: A total of 2,325,366 useable high-quality sequences were generated from the faecal samples analysed in this study and subjected to further bioinformatics analyses, which led to the identification of 128 OTUs and nine bacterial phyla, respectively. The phylum Firmicutes was predominant in all samples analysed (mean of 53.0%), followed by the phyla Proteobacteria (13.8%), Actinobacteria (13.7%) and Bacteroidetes (10.1%). Among others, bacteria of the order Lactobacillales, the family Enterococcaceae and genera Enterococcus and Dorea showed a trend towards increased abundance in Tc+ compared with Tc- samples, while no significant differences in OTU richness and diversity were recorded between Tc+ and Tc- samples (P = 0.485 and P = 0.581, respectively). However, Canonical Correlation and Redundancy Analyses were able to separate samples by infection status (P = 0.030 and P = 0.015, respectively), whic
- Published
- 2016
8. Vertical transmission of Anaplasma platys and Leishmania infantum in dogs during the first half of gestation
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Latrofa, MS, Dantas-Torres, F, de Caprariis, D, Cantacessi, C, Capelli, G, Lia, RP, Breitschwerdt, EB, Otranto, D, Latrofa, MS, Dantas-Torres, F, de Caprariis, D, Cantacessi, C, Capelli, G, Lia, RP, Breitschwerdt, EB, and Otranto, D
- Abstract
BACKGROUND: Leishmania infantum is a canine zoonotic vector-borne protozoan pathogen transmitted by phlebotomine sand flies, whereas Anaplasma platys is a bacterium most likely transmitted by ticks. While vertical transmission of L. infantum from pregnant bitches to their offspring has been documented, thus far no studies have explored the possibility of vertical transmission of A. platys in dogs. This study investigated the occurrence of vertical transmission of L. infantum and A. platys in sheltered dogs during the first half of gestation, in an area of southern Italy characterised by a high incidence of infection by both pathogens. METHODS: The study population included 20 bitches (n = 10 pregnant, at 25-35 days of pregnancy; n = 10 non-pregnant), all subjected to ovariohysterectomy, which were examined for the presence of L. infantum and A. platys via cytological screening of bone marrow and whole blood samples. Infection by L. infantum and A. platys was also tested by immunofluorescence antibody test (IFAT) and quantitative real-time PCR (qPCR) targeting both pathogens. Selected tissue samples (n = 210) collected during surgical procedures from bitches and foetuses (n = 20) were assessed for the presence of L. infantum and A. platys by qPCR targeting a fragment of the kinetoplast minicircle DNA (kDNA) and the 16S rRNA gene, respectively. RESULTS: Leishmania infantum DNA was not amplified from either uteri or ovaries from pregnant bitches or foetal tissue samples, whereas a subset of ovarian (n = 2) and uterine (n = 4) tissue samples from non-pregnant bitches were infected, with parasite loads of up to 3.09 × 10 and 7.51 parasite/PCR reaction, respectively. Conversely, uterine (n = 10) and ovarian (n = 8) tissues from both pregnant and non-pregnant bitches, together with a subset (n = 5) of foetal tissue samples were qPCR positive for A. platys. Leishmania infantum and A. platys nucleic acids were amplified from two uteri from non-pregnant bitches, with parasite
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- 2016
9. Angiostrongylus vasorum in the eye: new case reports and a review of the literature
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Colella, V, Lia, RP, Premont, J, Gilmore, P, Cervone, M, Latrofa, MS, D'Anna, N, Williams, D, Otranto, D, Colella, V, Lia, RP, Premont, J, Gilmore, P, Cervone, M, Latrofa, MS, D'Anna, N, Williams, D, and Otranto, D
- Abstract
BACKGROUND: Nematodes of the genus Angiostrongylus are important causes of potentially life-threatening diseases in several animal species and humans. Angiostrongylus vasorum affects the right ventricle of the heart and the pulmonary arteries in dogs, red foxes and other carnivores. The diagnosis of canine angiostrongylosis may be challenging due to the wide spectrum of clinical signs. Ocular manifestations have been seldom reported but have serious implications for patients. METHODS: The clinical history of three cases of infection with A. vasorum in dogs diagnosed in UK, France and Italy, was obtained from clinical records provided by the veterinary surgeons along with information on the diagnostic procedures and treatment. Nematodes collected from the eyes of infected dogs were morphologically identified to the species level and molecularly analysed by the amplification of the nuclear 18S rRNA gene. RESULTS: On admission, the dogs were presented with various degrees of ocular discomfort and hyphema because of the presence of a motile object in the eye. The three patients had ocular surgery during which nematodes were removed and subsequently morphologically and molecularly identified as two adult males and one female of A. vasorum. CONCLUSIONS: Three new cases of canine ocular angiostrongylosis are reported along with a review of other published clinical cases to improve the diagnosis and provide clinical recommendation for this parasitic condition. In addition, the significance of migratory patterns of larvae inside the host body is discussed. Veterinary healthcare workers should include canine angiostrongylosis in the differential diagnosis of ocular diseases.
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- 2016
10. Molecular dection of Eucoleus aerophilus in naturally infected dogs and cats
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DI CESARE, Angela, Castagna, G, Otranto, D, Meloni, S, Latrofa, Ms, Paoletti, Barbara, and Traversa, Donato
- Published
- 2012
11. Occurence of Giardia duodenalis sub-assemblage in horses
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Paoletti, Barbara, Otranto, D, Milillo, P, Latrofa, Ms, Giangaspero, A, Meloni, S, DI CESARE, Angela, and Traversa, Donato
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- 2012
12. Morphological and molecular data on the dermalmicrofilariae of a species of Cercopithifilaria from a dog in Sicily
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Otranto, D, Brianti, Emanuele, Dantas Torres, F, Weigl, S, Latrofa, Ms, Gaglio, Gabriella, Cauquil, L, Giannetto, Salvatore, and Bain, O.
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- 2011
13. Prevention of Canine Leishmaniosis in a Hyper-Endemic Area Using a Combination of 10% Imidacloprid/4.5% Flumethrin
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Ralph, SA, Otranto, D, Dantas-Torres, F, de Caprariis, D, Di Paola, G, Tarallo, VD, Latrofa, MS, Lia, RP, Annoscia, G, Breitshwerdt, EB, Cantacessi, C, Capelli, G, Stanneck, D, Ralph, SA, Otranto, D, Dantas-Torres, F, de Caprariis, D, Di Paola, G, Tarallo, VD, Latrofa, MS, Lia, RP, Annoscia, G, Breitshwerdt, EB, Cantacessi, C, Capelli, G, and Stanneck, D
- Abstract
BACKGROUND: Dogs are the main reservoir hosts of Leishmania infantum, the agent of human zoonotic visceral leishmaniosis. This study investigated the efficacy of a polymer matrix collar containing a combination of 10% imidacloprid and 4.5% flumethrin as a novel prophylactic measure to prevent L. infantum infections in young dogs from a hyper-endemic area of southern Italy, with a view towards enhancing current control strategies against both human and canine leishmaniosis. METHODOLOGY/PRINCIPAL FINDINGS: The study was carried out on 124 young dogs, of which 63 were collared (Group A) while 61 were left untreated (Group B), from March-April 2011 until March 2012. Blood and skin samples were collected at baseline (April 2011) and at the first, second, third and fourth follow-up time points (July, September 2011 and November 2011, and March 2012, respectively). Bone marrow and conjunctiva were sampled at baseline and at the fourth follow-up. Serological, cytological and molecular tests were performed to detect the presence of L. infantum in the different tissues collected. At the end of the trial, no dog from Group A proved positive for L. infantum at any follow-up, whereas 22 dogs from Group B were infected (incidence density rate = 45.1%); therefore, the combination of 10% imidacloprid and 4.5% flumethrin was 100% efficacious for the prevention of L. infantum infection in young dogs prior to their first exposure to the parasite in a hyper-endemic area for CanL. CONCLUSIONS: The use of collars containing 10% imidacloprid and 4.5% flumethrin conferred long-term protection against infection by L. infantum to dogs located in a hyper-endemic area, thus representing a reliable and sustainable strategy to decrease the frequency and spread of this disease among the canine population which will ultimately result in the reduction of associated risks to human health.
- Published
- 2013
14. Efficacy of an imidacloprid/flumethrin collar against fleas, ticks and tick-borne pathogens in dogs
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Dantas-Torres, F, Capelli, G, Giannelli, A, Ramos, RAN, Lia, RP, Cantacessi, C, de Caprariis, D, De Tommasi, AS, Latrofa, MS, Lacasella, V, Tarallo, VD, Di Paola, G, Qurollo, B, Breitschwerdt, E, Stanneck, D, Otranto, D, Dantas-Torres, F, Capelli, G, Giannelli, A, Ramos, RAN, Lia, RP, Cantacessi, C, de Caprariis, D, De Tommasi, AS, Latrofa, MS, Lacasella, V, Tarallo, VD, Di Paola, G, Qurollo, B, Breitschwerdt, E, Stanneck, D, and Otranto, D
- Abstract
BACKGROUND: Tick-borne diseases comprise a group of maladies that are of substantial medical and veterinary significance. A range of tick-borne pathogens, including diverse species of bacteria and protozoa, can infect both dogs and humans. Hence, the control of tick infestations is pivotal to decrease or prevent tick-borne pathogen transmission. Therefore, different commercial products with insecticidal, repellent or both properties have been developed for use on dogs. Recently, a collar containing a combination of imidacloprid 10% and flumethrin 4.5% has proven effective to prevent tick and flea infestations in dogs under field conditions and the infection by some vector-borne pathogens they transmit under laboratory-controlled conditions. METHODS: From March 2011 to April 2012, a field study was conducted in a private shelter in southern Italy to assess the efficacy of the imidacloprid/flumethrin collar against tick and flea infestations and to determine if this strategy would decrease tick-borne pathogen transmission in young dogs. A total of 122 animals were enrolled in the study and randomly assigned to group A (n = 64; collared) or group B (n = 58; untreated controls). Dogs were examined monthly for ticks and fleas and systematically tested for selected tick-borne pathogens. RESULTS: Compared to controls, the collar provided overall efficacies of 99.7% and 100% against tick and flea infestation, respectively. The overall efficacy for the prevention of tick-borne pathogens (i.e., Anaplasma platys and Babesia vogeli) was 91.6%. CONCLUSIONS: This study demonstrates that the imidacloprid/flumethrin collar is efficacious against flea and tick infestation as well as tick-borne pathogen transmission to dogs under field conditions.
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- 2013
15. Lungworms and gastrointestinal parasites of domestic cats: a European perspective
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Giannelli, A, Capelli, G, Joachim, A, Hinney, B, Losson, B, Kirkova, Z, René-Martellet, M, Papadopoulos, E, Farkas, R, Napoli, E, Brianti, E, Tamponi, C, Varcasia, A, Margarida Alho, A, Madeira De Carvalho, L, Cardoso, L, Maia, C, Mircean, V, Mihalca, AD, Miró, G, Schnyder, M, Cantacessi, C, Colella, V, Cavalera, MA, Latrofa, MS, Annoscia, G, Knaus, M, Halos, L, Beugnet, F, and Otranto, D
- Subjects
treatment ,diagnosis ,cats ,lungworms ,epidemiology ,Broadline® ,gastrointestinal parasites ,3. Good health - Abstract
With the exception of Aelurostrongylus abstrusus, feline lungworms have been poorly studied. Information on their distribution is patchy and mostly limited to case reports. In this study, the occurrence of feline lungworms and co-infecting gastrointestinal parasites has been investigated in 12 European countries (i.e. Austria, Belgium, Bulgaria, France, Greece, Hungary, Italy, Portugal, Romania, Spain, Switzerland and the United Kingdom). An average of 10 domestic cats, with regular outdoor access, was sampled each month for 12 months, and freshly passed faeces were collected. Stools were processed using a McMaster assay and a quantitative Baermann-Wetzel method. Animals positive for lungworms and/or gastrointestinal parasites were treated with a formulation containing fipronil, (S)-methoprene, eprinomectin, and praziquantel (Broadline®, Merial), and re-sampled 28 days post-treatment. The association between lungworm infection and risk factors was analysed using statistical medians/means and the efficacy of the treatment against each lungworm species was assessed. Of 1990 cats sampled, 613 (30.8%) were positive for at least one parasite, while 210 (10.6%) were infected by lungworms. The prevalence of lungworm infection varied between the sampled sites, with the highest recorded in Bulgaria (35.8%) and the lowest in Switzerland (0.8%). None of the cats from Austria or the United Kingdom were infected by lungworms. Aelurostrongylus abstrusus was the species most frequently detected (78.1%), followed by Troglostrongylus brevior (19.5%), Eucoleus aerophilus (14.8%) and Oslerus rostratus (3.8%). The overall efficacy of the treatment was 99% for A. abstrusus and 100% for T. brevior, O. rostratus and E. aerophilus. Data presented provide a comprehensive account of the diagnosis, epidemiology and treatment of feline lungworms in Europe, as well as of the occurrence of co-infections by gastrointestinal parasites.
16. Helminth infections and gut microbiota - a feline perspective
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Duarte, AM, Jenkins, TP, Latrofa, MS, Giannelli, A, Papadopoulos, E, De Carvalho, LM, Nolan, MJ, Otranto, D, and Cantacessi, C
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2. Zero hunger ,Lactobacilli ,Cat ,Microbial richness and diversity ,Gut microbiota ,16S rRNA ,Toxocara cati ,3. Good health - Abstract
$\textbf{BACKGROUND}$: Investigations of the relationships between the gut microbiota and gastrointestinal parasitic nematodes are attracting growing interest by the scientific community, driven by the need to better understand the contribution of parasite-associated changes in the composition of the gut flora to both host malnutrition and immune modulation. These studies have however been carried out mainly in humans and experimental animals, while knowledge of the make-up of the gut commensal flora in presence or absence of infection by parasitic nematodes in domestic animals is limited. In this study, we investigate the qualitative and quantitative impact that infections by a widespread parasite of cats (i.e. $\textit{Toxocara cati}$) exert on the gut microbiota of feline hosts. $\textbf{METHODS}$: The faecal microbiota of cats with patent infection by $\textit{T. cati}$ (= $\textit{Tc+}$), as well as that of negative controls (= $\textit{Tc-}$) was examined via high-throughput sequencing of the V3-V4 hypervariable region of the bacterial 16S rRNA gene, followed by bioinformatics and biostatistical analyses of sequence data. $\textbf{RESULTS}$: A total of 2,325,366 useable high-quality sequences were generated from the faecal samples analysed in this study and subjected to further bioinformatics analyses, which led to the identification of 128 OTUs and nine bacterial phyla, respectively. The phylum Firmicutes was predominant in all samples analysed (mean of 53.0%), followed by the phyla Proteobacteria (13.8%), Actinobacteria (13.7%) and Bacteroidetes (10.1%). Among others, bacteria of the order Lactobacillales, the family $\textit{Enterococcaceae}$ and genera $\textit{Enterococcus}$ and $\textit{Dorea}$ showed a trend towards increased abundance in $\textit{Tc+}$ compared with $\textit{Tc-}$ samples, while no significant differences in OTU richness and diversity were recorded between $\textit{Tc+}$ and $\textit{Tc-}$ samples ($\textit{P}$=0.485 and $\textit{P}$=0.581, respectively). However, Canonical Correlation and Redundancy Analyses were able to separate samples by infection status ($\textit{P}$=0.030 and $\textit{P}$=0.015, respectively), which suggests a correlation between the latter and the composition of the feline faecal microbiota. $\textbf{CONCLUSIONS}$: In spite of the relatively small number of samples analysed, subtle differences in the composition of the gut microbiota of $\textit{Tc+}$ vs $\textit{Tc-}$ cats could be identified, some of which in accordance with current data from humans and laboratory animal hosts. Nevertheless, the findings from this study contribute valuable knowledge to the yet little explored area of parasite-microbiota interactions in domestic animals.
17. Molecular detection of Colpodella sp. and other tick-borne pathogens in ticks of ruminants, Italy.
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Jimale KA, Bezerra-Santos MA, Mendoza-Roldan JA, Latrofa MS, Baneth G, and Otranto D
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- Animals, Italy epidemiology, Cattle, DNA, Protozoan genetics, Tick-Borne Diseases epidemiology, Tick-Borne Diseases microbiology, Tick-Borne Diseases parasitology, Tick-Borne Diseases veterinary, Rickettsia isolation & purification, Rickettsia genetics, Rickettsia classification, Babesia isolation & purification, Babesia genetics, Babesia classification, Rhipicephalus microbiology, Rhipicephalus parasitology, Theileria genetics, Theileria isolation & purification, Theileria classification, Sequence Analysis, DNA, Cattle Diseases parasitology, Cattle Diseases epidemiology, Cattle Diseases microbiology, Eucoccidiida genetics, Eucoccidiida isolation & purification, Eucoccidiida classification, Molecular Sequence Data, Ticks microbiology, Ticks parasitology, Phylogeny, Goats parasitology
- Abstract
Colpodella species are close relatives of Apicomplexan protozoa. Although most species of this genus are free-living organisms that feed on other protists and algae, reports indicate their occurence in ticks and human patients, including an individual with a history of tick bite manifesting neurological symptoms. During an investigation of tick-borne pathogens (TBPs) in blood samples of cattle, goats, and in ticks collected on them, Colpodella sp. DNA was detected in a Rhipicephalus bursa tick collected from cattle, while of Theileria sergenti/buffeli/orientalis, Babesia bigemina, Sarcocystis cruzi, Babesia spp., and Rickettsia spp. were molecularly detected in cattle, goats, and ticks in southern Italy. Data herein reported highlight the unprecedented presence of Colpodella sp. in ticks in Italy, raising concern due to the potential pathogenic role of this less known protozoan. This finding advocates for performing routine epidemiological surveys to monitor potential emerging vector-borne pathogens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)
- Published
- 2024
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18. Q3 lab-on-chip real-time PCR for the diagnosis of Leishmania infantum infection in dogs.
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Latrofa MS, Cereda M, Louzada-Flores VN, Dantas-Torres F, and Otranto D
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- Dogs, Animals, Lab-On-A-Chip Devices, Molecular Diagnostic Techniques methods, DNA, Protozoan genetics, Leishmania infantum genetics, Leishmania infantum isolation & purification, Real-Time Polymerase Chain Reaction methods, Dog Diseases diagnosis, Dog Diseases parasitology, Leishmaniasis, Visceral diagnosis, Leishmaniasis, Visceral veterinary, Leishmaniasis, Visceral parasitology, Sensitivity and Specificity
- Abstract
Leishmaniasis is a vector-borne disease caused by many Leishmania spp. which infect humans and other mammalian hosts. Leishmania infantum is the main agent of canine leishmaniasis (CanL) whose diagnosis is usually confirmed by serological and molecular tests. This study aimed to evaluate the clinical and analytical sensitivities of a lab-on-chip (LOC) real-time PCR applied on the portable Q3-Plus V2 platform (Q3 qPCR) in the detection of L. infantum . The Q3 qPCR performance was assessed by comparing the quantification cycle (Cq) values with those obtained using the qPCR run on a CFX96 Real-Time System (CFX96 qPCR). A total of 173 DNA samples (extracted from bone marrow, lymph node, blood, buffy coat, conjunctival swab, and skin) as well as 93 non-extracted samples (NES) (bone marrow, lymph node, blood, and buffy coat) collected from dogs were tested with both systems. Serial dilutions of each representative DNA and NES sample were used to assess the analytical sensitivity of the Q3 qPCR system. Overlapping Cq values were obtained with the Q3 qPCR and CFX96 qPCR, both using DNA extracted from L. infantum promastigotes (limit of detection, <1 promastigote per milliliter) and from biological samples as well as with NES. However, the Q3 qPCR system showed a higher sensitivity in detecting L. infantum in NES as compared with the CFX96 qPCR. Our data indicate that the Q3 qPCR system could be a reliable tool for detecting L. infantum DNA in biological samples, bypassing the DNA extraction step, which represents an advance in the point-of-care diagnostic of CanL., Competing Interests: The authors declare no conflict of interest.
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- 2024
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19. Development of an indirect ELISA for the serodiagnosis of canine infection by Onchocerca lupi.
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Latrofa MS, Louzada-Flores VN, Maia C, Kelly MA, Verocai GG, Cantacessi C, and Otranto D
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- Animals, Dogs, Cats, Onchocerca, Enzyme-Linked Immunosorbent Assay veterinary, Serologic Tests veterinary, Peptides, Cat Diseases, Dog Diseases diagnosis, Dog Diseases parasitology
- Abstract
Onchocerca lupi is a zoonotic filarioid parasite of dogs and cats with widespread distribution. A specific non-invasive diagnostic assay for the detection of O. lupi infections remains unavailable. This study aimed to assess the accuracy, specificity, and sensitivity of an ELISA test designed using nine peptides from two O. lupi proteins. Sera (n = 54) collected from O. lupi infected dogs from endemic areas (Portugal and USA), alongside sera from dogs positive for Dirofilaria immitis, D. repens, Cercopithifilaria bainae, and Acanthocheilonema reconditum (n = 53) from a non-endemic area for O. lupi, as well as from helminth-free dogs (n = 60), were tested. The checkerboard titration method was applied for the optimization of peptide concentrations and conjugate anti-dog dilutions. Sensitivity, specificity, and optimal cut-off values were calculated using ROC curve analysis. All peptides reacted against sera of O. lupi, with no correlation between optic density (OD) values and microfilariae (mfs) loads. Sensitivity and specificity values ranging from 85.45 to 100%, and 88.89% to 100%, respectively, were recorded for all peptides examined, with 100% specificity and sensitivity observed for peptides 40_3, 40_5, 130_3, 120_3 and 40_1, 130_5, respectively. The maximum cut-off value was observed for peptides 40_5 (0.765) and 40_3 (0.708). Testing of sera from dogs positive for other filarioids resulted in lower OD values (up to 1.565) for peptides 40_3 and 40_5 when compared with O. lupi (up to 2.929). The availability of this assay will be of value in epidemiological studies of canine O. lupi infection in both endemic and non-endemic areas, and in assessing the risk for zoonotic transmission., (© 2024. The Author(s).)
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- 2024
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20. Intracellular persistence of Leishmania tarentolae in primary canine macrophage cells.
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Louzada-Flores VN, Latrofa MS, Lucente MS, Dambrós BP, Mendoza-Roldan JA, Varotto-Boccazzi I, Cattaneo GM, Späth GF, Buonavoglia A, and Otranto D
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- Dogs, Animals, Macrophages parasitology, Monocytes, Leishmania infantum, Dog Diseases parasitology, Leishmaniasis, Visceral veterinary, Leishmaniasis, Visceral parasitology
- Abstract
Leishmania tarentolae is a non-pathogenic species first isolated from geckoes in the Mediterranean basin. The finding that dogs test positive against both Leishmania infantum and L. tarentolae raises questions regarding the ability of the latter species to persist and adapt to new hosts. This study aimed to evaluate in vitro the capability of L. tarentolae to colonize, survive and persist in canine primary monocyte-derived mononuclear cells. Monocytes were isolated from dog whole blood samples and placed in 24-well plates for differentiation into macrophages and for incubation with L. tarentolae field-isolated strains (RI-325 and SF-178) and laboratory (LEM-124) strain; the parasite burden was assessed at different time points post-infection. The L. infantum laboratory strain (MON-1) was used as control. Infection parameters were evaluated by microscopy, counting the number of amastigotes/200 infected cells, and by duplex real-time PCR from supernatants and detached cells. Similar to L. infantum, L. tarentolae strains developed into round-shaped amastigote-like forms, with higher infection rates detected at 4 h followed by an overall decrease until 48 h. RI-325 presented also a higher infection rate at 72 h. Data showed that L. tarentolae strains infect and persist inside in vitro primary canine mononuclear cells, opening new perspectives for further laboratory studies., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)
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- 2023
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21. Co-infection patterns of vector-borne zoonotic pathogens in owned free-ranging dogs in central Chile.
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Cevidanes A, Di Cataldo S, Muñoz-San Martín C, Latrofa MS, Hernández C, Cattan PE, Otranto D, and Millán J
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- Dogs, Animals, Humans, Chile epidemiology, Coinfection epidemiology, Coinfection veterinary, Dog Diseases epidemiology
- Abstract
We investigated the co-occurrence of the nine of the most relevant canine vector-borne pathogens (CVBP) using conventional and real-time PCR and evaluated risk factors and potential non-apparent haematological alterations associated with co-infection in 111 rural, owned, free-ranging dogs in the Metropolitan Region of Chile.At least one pathogen was detected in 75% of the dogs. DNA of Anaplasma platys (Ap; 36%), Candidatus Mycoplasma haematoparvum (CMhp; 31%), Mycoplasma haemocanis (Mhc; 28%), Trypanosoma cruzi (17%), Leishmania spp. (4.5%), and Acanthocheilonema reconditum (1%) was detected. All dogs were negative for Ehrlichia spp., Rickettsia spp., Bartonella spp., Piroplasmida, and Hepatozoon spp. Thirty-eight dogs (34%) were coinfected. CMhp was involved in 71%, Mhc in 58%, and Ap in 50% of the co-infections. The most common co-infection pattern was CMhp-Mhc (37% of the cases). The prevalence of Ap was higher in juvenile than in adult dogs, whereas the opposite was found for CMhp and Mhc. Adult dogs were four times more likely of being co-infected than juveniles. Co-infected animals showed higher white blood cell count, segmented neutrophil count, and GGT levels than non-co-infected dogs. Clinically healthy but infected dogs may act as reservoirs of CVBP, and their free-ranging behavior would facilitate the spread of these pathogens to other dogs as well as human beings or wild carnivores., (© 2022. The Author(s).)
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- 2023
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22. In vitro maintenance of the endosymbiont Wolbachia of Dirofilaria immitis.
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Manoj RRS, Latrofa MS, Louni M, Laidoudi Y, Fenollar F, Otranto D, and Mediannikov O
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- Animals, Dogs, Microfilariae, Dirofilaria immitis, Dirofilariasis, Wolbachia, Dog Diseases microbiology
- Abstract
Wolbachia has an obligatory mutualistic relationship with many onchocercid nematodes of the subfamilies Dirofilariinae and Onchocercinae. Till date, no attempts have been made for the in vitro cultivation of this intracellular bacterium from the filarioid host. Hence, the current study attempted cell co-culture method using embryonic Drosophila S2 and the LD cell lines to cultivate Wolbachia from Dirofilaria immitis microfilariae (mfs) harvested from infected dogs. Microfilariae (mfs = 1500) were inoculated in shell vials supplemented with Schneider medium using both cell lines. The establishment and multiplication of the bacterium were observed during the initial inoculation, at day 0 and before every medium change (from days 14 to 115). An aliquot (50 µl) from each time point was tested by quantitative real-time PCR (qPCR). Comparing the average of Ct values, obtained by the tested parameters (i.e., LD/S2 cell lines and mfs with/without treatment), the S2 cell line without mechanical disruption of mfs provided the highest Wolbachia cell count by qPCR. Despite the maintenance of Wolbachia within both S2 and LD-based cell co-culture models for up to 115 days, a definitive conclusion is still far. Further trials using fluorescent microscopy and viable staining will help to demonstrate the cell line infection and viability of Wolbachia. Use of considerable amount of untreated mfs to inoculate the Drosophilia S2 cell lines, as well as the supplementation of the culture media with growth stimulants or pre-treated cells to increase their susceptibility for the infection and development of a filarioid-based cell line system are recommended for the future trials., (© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
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- 2023
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23. Interaction between Wolbachia pipientis and Leishmania infantum in heartworm infected dogs.
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Latrofa MS, Varotto-Boccazzi I, Louzada-Flores VN, Iatta R, Mendoza-Roldan JA, Roura X, Zatelli A, Epis S, Bandi C, and Otranto D
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- Female, Animals, Dogs, Cytokines, Dirofilaria immitis, Wolbachia, Leishmania infantum
- Abstract
Background: Wolbachia is a Gram-negative endosymbiont associated with several species of arthropods and filarioid nematodes, including Dirofilaria immitis. This endosymbiont may elicit a Th1 response, which is a component of the immunity against Leishmania infantum., Methods: To investigate the interactions between Wolbachia of D. immitis and L. infantum in naturally infected dogs and cytokine circulation, dogs without clinical signs (n = 187) were selected. Dogs were tested for microfilariae (mfs) by Knott, for female antigens of D. immitis by SNAP, and for anti-L. infantum antibodies by IFAT and assigned to four groups. Dogs of group 1 (G1) and 2 (G2) were positive for D. immitis and positive or negative to L. infantum, respectively. Dogs of group 3 (G3) and 4 (G4) were negative to D. immitis and positive or negative to L. infantum, respectively. Wolbachia and L. infantum DNA was quantified by real-time PCR (qPCR) in dog blood samples. A subset of dogs (n = 65) was examined to assess pro- and anti-inflammatory cytokine production using an ELISA test., Results: Of 93 dogs positive to D. immitis with circulating mfs, 85% were positive to Wolbachia, with the highest amount of DNA detected in G1 and the lowest in dogs with low mfs load in G1 and G2. Among dogs positive to L. infantum, 66% from G1 showed low antibody titer, while 48.9% from G3 had the highest antibody titer. Of 37 dogs positive to Wolbachia from G1, 26 (70.3%) had low antibody titers to L. infantum (1:160). Among cytokines, TNFα showed the highest mean concentration in G1 (246.5 pg/ml), IFNγ being the one most represented (64.3%). IL-10 (1809.5 pg/ml) and IL-6 (123.5 pg/ml) showed the highest mean concentration in dogs from G1. A lower percentage of dogs producing IL-4 was observed in all groups examined, with the highest mean concentration (2794 pg/ml) recorded in G2., Conclusion: Results show the association of D. immitis and Wolbachia with the lower antibody titers of L. infantum in co-infected dogs, suggesting the hypothesis that the endosymbiont may affect the development of the patent leishmaniosis. However, due to the limitations associated with the heterogeneity of naturally infected dogs in field conditions, results should be validated by investigation on experimental models., (© 2023. The Author(s).)
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- 2023
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24. Leishmania spp. in Squamata reptiles from the Mediterranean basin.
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Mendoza-Roldan JA, Latrofa MS, Tarallo VD, Manoj RR, Bezerra-Santos MA, Annoscia G, Iatta R, and Otranto D
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- Animals, DNA, Kinetoplast genetics, Dogs, Snakes genetics, Dog Diseases epidemiology, Leishmania infantum genetics, Leishmaniasis epidemiology, Leishmaniasis veterinary, Lizards genetics
- Abstract
Leishmania tarentolae is a nonpathogenic trypanosomatid species isolated from geckoes in the Mediterranean basin. In Italy, L. tarentolae and Leishmania infantum occur in sympatry in areas where canine leishmaniosis (CanL) is endemic. Here, we investigated the prevalence and distribution of L. tarentolae in reptilian hosts and the presence of L. infantum in the same animals in southern Italy. From April 2020 to April 2021, lizards and snakes were captured in urban, peri-urban sites, natural parks, and dog shelters. Blood was individually sampled and assessed for the presence of Leishmania spp. by cytology and/or molecular biology. Captured lizards were euthanized and their tissues/organs were tested by duplex real-time-PCR for the detection of L. infantum and L. tarentolae and the positive ones were amplified by conventional PCR to confirm the species identification by sequencing. Samples were also screened by real-time PCR for L. infantum kDNA minicircle. Of the 259 collected Squamata reptiles, 34 (13.1%) specimens scored positive for Leishmania spp., with a prevalence of 10% for L. tarentolae (n = 26) and of 3.1% for L. infantum (n = 8). Positive lizards were mostly Podarcis siculus (n = 26) from dog shelters, with six specimens positive for L. infantum. In addition, L. tarentolae was detected in one Podarcis filfolensis lizard, and in five T. mauritanica geckos (with one gecko and P. filfolensis lizard showing amastigote-like forms in thrombocytes), whereas L. infantum was detected in one T. mauritanica, and one Hemidactylus turcicus. Data herein reported demostrate that saurian reptiles living in CanL endemic areas of the Mediterranean basin may be exposed to both L. tarentolae and L. infantum indicating a sympatric circulation of these species in different vertebrate hosts. Moreover, this study represents the first molecular evidence of L. infantum infection in European reptiles., (© 2022 Wiley-VCH GmbH.)
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- 2022
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25. Leishmania (Sauroleishmania) tarentolae isolation and sympatric occurrence with Leishmania (Leishmania) infantum in geckoes, dogs and sand flies.
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Mendoza-Roldan JA, Zatelli A, Latrofa MS, Iatta R, Bezerra-Santos MA, Annoscia G, Gernone F, Votýpka J, Modrý D, Tichá L, Volf P, and Otranto D
- Subjects
- Animals, Dogs, Female, Humans, Polymerase Chain Reaction, Leishmania infantum genetics, Lizards, Phlebotomus parasitology, Psychodidae parasitology
- Abstract
The trypanosomatid protist Leishmania tarentolae is a saurian-associated parasite vectored by the Sergentomyia minuta sand fly. This study aimed to confirm the circulation of L. infantum and L. tarentolae in sand flies, reptiles and dogs and to isolate new strains of these protists. Reptilian and sheltered dog blood samples were collected, and sand flies were captured. Samples were tested for Leishmania spp. using duplex real-time PCR (dqPCR) and real-time PCR (qPCR); the origin of blood meal was identified in engorged sand flies by conventional PCR. The reptilian blood and intestinal content of sand fly females were cultured. Dog sera were tested by IFAT using both Leishmania species. Four Tarentola mauritanica geckoes were molecularly positive for L. infantum or L. tarentolae, with no co-infections; moreover, amastigote-like forms of L. infantum were observed in the bone marrow. 24/294 sand flies scored positive for Leishmania spp. by dqPCR, 21 S. minuta and two Phlebotomus perniciosus were positive for L. tarentolae, while only a single Ph. perniciosus was positive for L. infantum. Blood meal analysis confirmed reptile and dog in S. minuta, dog and human in Ph. perniciosus and dog in Phlebotomus neglectus. Two axenic strains of L. tarentolae were obtained. Twelve of 19 dogs scored positive for L. infantum and L. tarentolae by IFAT and three of them also for L. infantum by dqPCR, and six by qPCR. These data confirm the sympatric circulation of L. infantum and L. tarentolae in geckoes, sand flies, and dogs, and suggest that geckoes may be infected with L. infantum., Competing Interests: The authors have declared that no competing interests exist.
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- 2022
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26. Treatment with doxycycline is associated with complete clearance of circulating Wolbachia DNA in Dirofilaria immitis-naturally infected dogs.
- Author
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Louzada-Flores VN, Kramer L, Brianti E, Napoli E, Mendoza-Roldan JA, Bezerra-Santos MA, Latrofa MS, and Otranto D
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- Animals, Dogs, Doxycycline pharmacology, Doxycycline therapeutic use, Cell-Free Nucleic Acids, Dirofilaria immitis genetics, Dirofilariasis drug therapy, Dog Diseases parasitology, Wolbachia genetics
- Abstract
Bacteria of the genus Wolbachia are endosymbionts of parasitic filarial nematodes, including Dirofilaria immitis, and are a target for the treatment of canine heartworm disease. In the present study, 53 naturally-infected dogs were divided in three groups, based on their positivity to D. immitis by antigen and Knott tests, to assess the efficacy of doxycycline treatment in eliminating Wolbachia from circulating blood. At T0, dogs that scored positive to both tests (G1) or to antigen only (G2) were submitted to doxycycline (10 mg/kg BID PO) treatment and to 10% Imidacloprid + 2.5% Moxidectin (Advocate®), while those negative to both tests (G3) received only 10% Imidacloprid + 2.5% Moxidectin (Advocate®). All dogs were followed-up for one year, monthly treated with Advocate® and regularly monitored by antigen and Knott tests. During the whole period, all blood samples were screened for Wolbachia-D. immitis DNA load by quantitative real-time PCR (qPCR). At T0, 88.2% of the microfilariemic dogs were positive for Wolbachia DNA, while none of the dogs from G2 or G3 were positive. Wolbachia DNA was no longer detectable in dogs from G1 following 1 month of doxycycline treatment and microfilariae (mfs) were cleared at T2. All dogs from the G1 and G2 were negative for D. immitis antigen at 12 months. Results of this study suggest that successful elimination of mfs by doxycycline is associated with complete clearance of Wolbachia DNA in D. immitis-naturally infected dogs., (Copyright © 2022. Published by Elsevier B.V.)
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- 2022
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27. Cercopithifilaria spp. in ticks of companion animals from Asia: new putative hosts and vectors.
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Bezerra-Santos MA, de Macedo LO, Nguyen VL, Manoj RR, Laidoudi Y, Latrofa MS, Beugnet F, and Otranto D
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- Animals, Cats, Dogs, Pets, Taiwan, Thailand, Cat Diseases epidemiology, Dog Diseases epidemiology, Filarioidea genetics, Ixodidae, Rhipicephalus sanguineus
- Abstract
Cercopithifilaria bainae, Cercopithifilaria grassi, and Cercopithifilaria sp. II sensu Otranto et al., 2013 tick borne filarioids are typically found in dogs. Among them, Cercopithifilaria bainae has a worldwide distribution according to the occurrence of its tick vector, Rhipicephalus sanguineus sensu lato (s.l.). Nevertheless, in Asian countries, despite the wide presence of this tick species, data on Cercopithifilaria spp. are scant. Therefore, this study aimed to assess the occurrence of these dermal filarioids in ixodid ticks collected on dogs and cats from Asian countries, providing a better epidemiological picture on their distribution in this continent. Ticks (n = 687) of the species Rhipicephalus sanguineus s. l. (n = 667), Rhipicephalus haemaphysaloides (n = 8), Haemaphysalis longicornis (n = 7), Haemaphysalis campanulata (n = 1), Haemaphysalis wellingtoni (n = 2), Haemaphysalis hystricis (n = 1), and Ixodes sp. (n = 1) were collected on dogs and cats under the frame of previous studies in China, India, Indonesia, Malaysia, Singapore, Taiwan, Thailand, the Philippines and Vietnam. Tick samples were molecularly screened for Cercopithifilaria spp. by conventional PCR and real-time PCR using two pair of primers targeting partial sequences of cytochrome c oxidase 1 gene. Overall, Cercophitifilaria spp. DNA was detected in 9.5% (n = 65/687) of the tick specimens tested, with C. bainae being the most prevalent species (8.9%), followed by C. grassii (0.6%). Most Cercophitifilaria spp. positive ticks were collected on dogs (92.3%; 60/65); whereas ticks collected on cats represented 7.7% of the positive specimens. In addition, Cercopithifilaria spp. were mostly detected in R. sanguineus s.l. ticks (96.9%; 63/65), followed by Rhipicephalus haemaphysaloides (3.1%; 2/65). Data herein presented demonstrate the occurrence of dermal tick borne filarioids of the genus Cercopithifilaria in several Asian countries, with C. bainae being the most prevalent species. We also report for the first time the molecular detection of C. bainae in R. sanguineus s.l. ticks collected on cats, as well as in R. haemaphysaloides ticks, suggesting that the biological cycle of this filarioid species may involve other intermediate and definitive hosts than R. sanguineus s.l. and dogs. However, confirmatory studies on the role of other tick species and domestic cats on the biology of C. bainae are advocated., (Copyright © 2022. Published by Elsevier GmbH.)
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- 2022
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28. Wild Boar (Sus scrofa) as Reservoir of Zoonotic Yeasts: Bioindicator of Environmental Quality.
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Rhimi W, Sgroi G, Aneke CI, Annoscia G, Latrofa MS, Mosca A, Veneziano V, Otranto D, Alastruey-Izquierdo A, and Cafarchia C
- Subjects
- Animals, Antifungal Agents pharmacology, Azoles, Candida, Candida albicans, Fluconazole, Microbial Sensitivity Tests, Sus scrofa, Swine, Yeasts genetics, Drug Resistance, Fungal, Environmental Biomarkers
- Abstract
Wildlife animals are recognized as reservoirs for zoonotic fungi and their faeces might play an important role in introducing pathogens into the environment. Thought wild boar (Sus scrofa) population has dramatically increased across Europe, information about their possible role in dissemination of zoonotic pathogenic yeasts in the environment is scant. Therefore, fecal samples (n = 124) from wild boars from Campania region (Southern Italy) were collected and yeasts identified biochemically and molecularly by sequencing of the internal transcribed spacer region and their phylogenetical relationship assessed. The antifungal susceptibility profiles of yeasts were also investigated using AFST-EUCAST method. Yeasts were isolated from 50.1% of the samples with the highest occurrence in samples from the province of Salerno (61.1%). A total of 368 Candida strains belonging to nine species were identified, with Candida albicans (45.7%), followed by Candida krusei (15.2%), Kazachstania slooffiae (9.8%) and Candida parapsilosis (7.6%) as the most prevalent identified species. Among C. albicans four sequence types (i.e., ST1-ST4) were identified with an intraspecific nucleotide difference up to 0.21%. The ML tree grouped all representative sequence types as paraphyletic clades with those of the references yeast species, respectively and supported by high bootstrap values. Fluconazole was the less active drug whereas, posaconazole, voriconazole, and isavuconazole the most active one. No resistance phenomena were observed for C. albicans and high MICs values for 5FC, azoles and echinocandines were registered in non-albicans Candida spp. This study showed, for the first time, the important role of wild boars in dissemination of pathogenic fungi in the environment. The absence of resistance phenomena in the Candida spp. might reflect environmental free from residues of azoles antifungals pollution or chemicals and suggests the role of wild boar as bio indicators of environment quality., (© 2022. The Author(s), under exclusive licence to Springer Nature B.V.)
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- 2022
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29. Genetic and geographical delineation of zoonotic vector-borne helminths of canids.
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Laidoudi Y, Bedjaoui S, Latrofa MS, Fanelli A, Dantas-Torres F, and Otranto D
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- Animals, Disease Vectors, Genetic Variation, Onchocerca, Phylogeny, Canidae, Dirofilaria immitis, Dirofilaria repens
- Abstract
Several zoonotic vector-borne helminths (VBHs) infesting canids cause serious veterinary and medical diseases worldwide. Increasing the knowledge about their genetic structures is pivotal to identify them and therefore to settle effective surveillance and control measures. To overcome the limitation due to the heterogeneity of large DNA sequence-datasets used for their genetic characterization, available cytochrome c oxidase subunit 1 (cox1) (n = 546) and the 12S rRNA (n = 280) sequences were examined using combined bioinformatic approach (i.e., distance-clustering, maximum likelihood phylogeny and phylogenetic evolutionary placement). Out of the 826 DNA available sequences from GenBank, 94.7% were characterized at the haplotype level regardless sequence size, completeness and/or their position. A total of 89 different haplotypes were delineated either by cox1 (n = 35), 12S rRNA (n = 21) or by both genes (n = 33), for 14 VBHs (e.g., Acanthocheilonema reconditum, Brugia spp., Dirofilaria immitis, Dirofilaria repens, Onchocerca lupi and Thelazia spp.). Overall, the present approach could be useful for studying global genetic diversity and phylogeography of VBHs. However, as barcoding sequences were restricted to two mitochondrial loci (cox1 and 12S rRNA), the haplotype delineation proposed herein should be confirmed by the characterization of other nuclear loci also to overcome potential limitations caused by the heteroplasmy phenomenon within the mitogenome of VBHs., (© 2022. The Author(s).)
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- 2022
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30. Onchocerca lupi in imported dogs in the UK: implications for animal and public health.
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McGarry JW, Carrozza R, Bradley C, Latrofa MS, Makepeace BL, and Otranto D
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- Animals, Dogs, Eye, Public Health, United Kingdom epidemiology, Dog Diseases diagnosis, Dog Diseases drug therapy, Dog Diseases surgery, Onchocerca genetics
- Abstract
Background: Onchocerca lupi is a filarial nematode affecting dogs, and occasionally cats and humans, in continental Europe, North Africa, the Middle East, and the USA. Adult worms are usually found in periocular nodules and enucleation is sometimes required if the infection fails to respond to other treatment options., Case Presentation: Here, we report the presence of O. lupi in the UK for the first time. Of two dogs re-homed from continental Europe, one developed an ocular nodule seven years after arrival from Portugal. The conjunctival perilimbal mass in its left eye was surgically removed but despite anthelminthic treatment, a further nodule developed in the same eye six months later. In the second case - a dog imported from Romania 12 months earlier - a perilimbal mass was excised from the left eye and prior anthelminthic treatment was supplemented with oral prednisolone and doxycycline. However, nodules recurred, and the left globe was subsequently enucleated. Conjunctival hyperaemia then appeared in the right eye and neither additional anthelminthic treatment nor removal of worm masses failed to prevent the further development of lesions. Excised adult worms were identified in both cases as O. lupi based on morphological characteristics, as well as PCR and sequencing of cytochrome c oxidase subunit I and 12S rRNA gene fragments., Conclusion: O. lupi parasitosis can apparently remain cryptic in dogs for several years before any clinical signs manifest. Moreover, the progression of infection can be highly aggressive and recalcitrant to both surgical intervention and anthelminthic treatment. Increasingly, former stray dogs of unknown infection status are entering the UK, raising both veterinary and public health concerns., (© 2022. The Author(s).)
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- 2022
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31. Molecular detection and characterization of the endosymbiont Wolbachia in the European hedgehog flea, Archaeopsylla erinacei.
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Manoj RRS, Latrofa MS, Bezerra-Santos MA, Sgroi G, Samarelli R, Mendoza-Roldan JA, and Otranto D
- Subjects
- Animals, Bacterial Outer Membrane Proteins analysis, Female, Italy, Male, Phylogeny, RNA, Bacterial analysis, RNA, Ribosomal, 16S analysis, Wolbachia classification, Wolbachia genetics, Siphonaptera microbiology, Symbiosis, Wolbachia isolation & purification
- Abstract
Wolbachia, the endosymbiont of arthropods and onchocercid nematodes is present in many medically important insect species, being also considered for the indirect control of parasitic ones. Archaeopsylla erinacei is a flea species infesting hedgehogs acting as vector of Rickettsia felis, Bartonella henselae, and Rickettsia helvetica, thus having public health relevance. The Wolbachia surface protein (wsp) and 16S rRNA genes were used to determine the presence, prevalence and molecular typing of Wolbachia in this flea species collected in two regions of southern Italy. Of the 45 fleas tested (n = 16 males, 35.6%; n = 29 females, 64.4%), 43 (95.6%; 95% CI: 84.8-99.2) scored positive for Wolbachia, of which 15 (33.3%) and 28 (62.2%) were males and females, respectively. The sex-wise prevalence of this endosymbiont was almost equal in both sexes (males 93.8%; 95% CI: 69.5-99.7; females 96.7%; 95% CI: 83.1-99.8). Single locus sequence analysis (SLST) of Wolbachia revealed two sequence types for 16S rRNA gene, named as wAr_15227 and wAr_15234, which came from two different areas, equally distributed in male and female fleas, whilst only one sequence type was identified for wsp gene. The phylogenetic analysis placed the two 16S rRNA sequence types in paraphyletic clades belonging to the supergroup A and B, respectively. Whilst, the tree of wsp gene clustered the corresponding sequence in the same clade including those of Wolbachia supergroup A. In MLST analyses, both Wolbachia sequence types clustered in a monophyletic clade with Drosophila nikananu (wNik) and Drosophila sturtevanti (wStv) from supergroup A. ClonalFrame analysis revealed a recombination event in the wAr_15234 strain which came from Apulia region. Scientific knowledge of the presence/prevalence of Wolbachia among medically important fleas, may contribute to develop an alternative biological method for the vector control., (Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.)
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- 2022
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32. Zoonotic Dirofilaria immitis and Dirofilaria repens infection in humans and an integrative approach to the diagnosis.
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Mendoza-Roldan JA, Gabrielli S, Cascio A, Manoj RRS, Bezerra-Santos MA, Benelli G, Brianti E, Latrofa MS, and Otranto D
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- Animals, Dog Diseases diagnosis, Dog Diseases epidemiology, Dogs, Humans, Islands, Italy, Phylogeny, Seroepidemiologic Studies, Dirofilaria immitis genetics, Dirofilaria repens genetics, Dirofilariasis diagnosis, Dirofilariasis epidemiology, Zoonoses diagnosis, Zoonoses parasitology
- Abstract
Dirofilariosis by Dirofilaria immitis and Dirofilaria repens is endemic in dogs from countries of the Mediterranean basin. Both species may infect humans, with most of the infected patients remaining asymptomatic. Based on the recent description of the southernmost hyperendemic European focus of heartworm disease in dogs from the Pelagie archipelagos, we performed a serological and molecular survey in human population of that area. Human blood samples were collected in the islands of Linosa (n=101) and Lampedusa (n=296) and tested by ELISA and molecular test for the detection of D. immitis and D. repens. Samples were also screened for filarioid-associated endosymbionts, Wolbachia sp. The seroprevalence of D. immitis and D. repens was, respectively, 7.9% and 3.96% in Linosa, and 7.77% and 19.93% in Lampedusa. Out of 397 human blood samples tested molecularly, 4 scored positive (1%) for Dirofilaria spp. by qPCR (i.e., three for D. immitis and one for D. repens) and 6 (1.5%) for Wolbachia. Of the qPCR positive for Dirofilaria spp., only D. repens was amplified by cPCR and was positive for Wolbachia. In the phylogenetic analysis, the sequence of Wolbachia detected in D. repens positive samples clustered along with other C supergroup filarioids. Our results overlap with the recent prevalence data collected on dogs from the same area, where D. immitis is prevalent in Linosa and D. repens prevails in Lampedusa. Molecular detection of D. immitis in human blood is quite unusual considering that humans are dead-end hosts for dirofilarial infection and most of the human cases described so far in Europe were ascribed to D. repens. An integrative diagnostic approach using serum analysis and Wolbachia detection is also presented. In endemic areas for canine dirofilarioses humans are exposed to the infection, suggesting the importance of One Health approach in diagnosing, treating and controlling this zoonotic parasitosis., (Copyright © 2021. Published by Elsevier B.V.)
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- 2021
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33. Molecular detection of zoonotic blood pathogens in ticks from illegally imported turtles in Italy.
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Manoj RRS, Mendoza-Roldan JA, Latrofa MS, Remesar S, Brianti E, and Otranto D
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- Anaplasma isolation & purification, Animals, Commerce, Ehrlichia isolation & purification, Female, Internationality, Male, Ticks microbiology, Ticks parasitology, Turtles parasitology
- Abstract
International trade of animals may represent a gateway for the spreading of zoonotic pathogens and their vectors. Amongst animals, reptiles are commonly illegally imported worldwide, being Italy in the fifth position of importation of these animals. Thus, the current study analysed the pathogens associated with Hyalomma aegyptium ticks, which were collected from illegally imported tortoises from North Africa to Italy. All tick DNA samples were tested by conventional PCR for the presence of Anaplasma spp., Babesia spp., Borrelia spp., Coxiella burnetti, Ehrlichia spp., Hepatozoon spp., Rickettsia spp. and microfilariae of filarioids. Out of 22% (n=161) of ticks screened, 78.9% (n=127) were males and 21.1% (n=34) females. Among them, three male specimens collected from two different turtles (1.9%; 95% CI; 0.5-5.5) scored positive for Anaplasma spp./Ehrlichia spp., whereas all females were negative. BLAST analysis of the sequences obtained from positive samples revealed 99-99.3% nucleotide identity with the sequence of Ehrlichia ewingii available in GenBank. The finding of E. ewingii in ticks from imported reptiles warrants the need for imposing strict rules in the international trade of reptiles to effectively reduce the introduction of exotic pathogens and their vectors in new geographic areas., (Copyright © 2021. Published by Elsevier B.V.)
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- 2021
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34. Trypanosoma ( Megatrypanum ) pestanai in Eurasian badgers ( Meles meles ) and Ixodidae ticks, Italy.
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Sgroi G, Iatta R, Paolo Lia R, Latrofa MS, Samarelli R, Camarda A, and Otranto D
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- Animals, Female, Humans, Ixodes, Ixodidae, Mustelidae parasitology, Siphonaptera, Trypanosoma genetics
- Abstract
Trypanosomes are haemoflagellate protozoa transmitted by blood-feeding arthropods causing infections in a wide range of mammals, including humans. Adult badgers (Meles meles, n = 2), displaying severe paralysis, ataxia and severe ectoparasite infestation, were rescued from a peri-urban area of Bari (southern Italy). Blood samples and ectoparasites were screened for Trypanosoma spp. by the combined PCR/sequencing approach, targeting a fragment of 18S rRNA gene. Smears of haemolymph, guts and salivary glands of the alive ticks were microscopically observed. No haematological alterations, except thrombocytopenia, were found. Trypomastigotes and epimastigotes were observed in the blood smears of both badgers and Trypanosoma pestanai was molecularly identified. Out of 33 ticks (i.e. n = 31 Ixodes canisuga, n = 2 Ixodes ricinus) and two fleas (Ctenocephalides felis), 11 specimens (n = 5 I. canisuga engorged nymphs, n = 4 engorged females and n = 2 I. ricinus engorged females) tested positive only for T. pestanai DNA. All smears from ticks were negative. The present study firstly revealed T. pestanai in Ixodidae and badgers from Italy, demonstrating the occurrence of the protozoan on the peninsula. Further studies are needed to clarify the occurrence of the only known vector of this parasite, Paraceras melis flea, as well as other putative arthropods involved in the transmission of T. pestanai.
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- 2021
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35. Leishmania tarentolae and Leishmania infantum in humans, dogs and cats in the Pelagie archipelago, southern Italy.
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Iatta R, Mendoza-Roldan JA, Latrofa MS, Cascio A, Brianti E, Pombi M, Gabrielli S, and Otranto D
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- Adult, Aged, Aged, 80 and over, Animals, Blotting, Western veterinary, Cat Diseases parasitology, Cats, Dog Diseases parasitology, Dogs, Enzyme-Linked Immunosorbent Assay veterinary, Female, Humans, Italy epidemiology, Leishmaniasis, Visceral diagnosis, Leishmaniasis, Visceral parasitology, Male, Middle Aged, Prevalence, Public Health, Real-Time Polymerase Chain Reaction veterinary, Serologic Tests, Sicily epidemiology, Surveys and Questionnaires, Young Adult, Cat Diseases epidemiology, Dog Diseases epidemiology, Leishmania infantum genetics, Leishmaniasis, Visceral epidemiology, Leishmaniasis, Visceral veterinary
- Abstract
Visceral leishmaniasis (VL) caused by Leishmania infantum is endemic in the Mediterranean basin with most of the infected human patients remaining asymptomatic. Recently, the saurian-associated Leishmania tarentolae was detected in human blood donors and in sheltered dogs. The circulation of L. infantum and L. tarentolae was investigated in humans, dogs and cats living in the Pelagie islands (Sicily, Italy) by multiple serological and molecular testing. Human serum samples (n = 346) were tested to assess the exposure to L. infantum by immunofluorescence antibody test (IFAT), enzyme-linked immunosorbent assay (ELISA) and Western blot (WB) and to L. tarentolae by IFAT. Meanwhile, sera from dogs (n = 149) and cats (n = 32) were tested for both Leishmania species by IFAT and all blood samples, including those of humans, by specific sets of real time-PCR for L. infantum and L. tarentolae. The agreement between serological tests performed for human samples, and between serological and molecular diagnostic techniques for both human and animal samples were also assessed. Overall, 41 human samples (11.8%, 95% CI: 8.9-15.7) were positive to L. infantum (5.2%, 95% CI: 3.3-8.1), L. tarentolae (5.2%, 95% CI: 3.3-8.1) and to both species (1.4%, 95% CI: 0.6-3.3) by serology and/or molecular tests. A good agreement among the serological tests was determined. Both Leishmania spp. were serologically and/or molecularly detected in 39.6% dogs and 43.7% cats. In addition to L. infantum, also L. tarentolae circulates in human and animal populations, raising relevant public health implications. Further studies should investigate the potential beneficial effects of L. tarentolae in the protection against L. infantum infection., Competing Interests: The authors have declared that no competing interests exist.
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- 2021
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36. Case Report: A Human Case of Onchocerca lupi Mimicking Nodular Scleritis.
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Hasanreisoglu M, Halim MS, Latrofa MS, Mendoza-Roldan JA, Ogut B, Yilmaz M, Tefon Aribas AB, Caglar K, Gonul II, Dogruman-Al F, Nguyen QD, and Otranto D
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- Diagnosis, Differential, Female, Humans, Oculomotor Muscles surgery, Onchocerciasis, Ocular pathology, Onchocerciasis, Ocular surgery, Turkey, Young Adult, Oculomotor Muscles pathology, Onchocerciasis, Ocular diagnosis, Scleritis diagnosis
- Abstract
We report a case of human ocular onchocerciasis by zoonotic Onchocerca lupi presenting as nodular scleritis. Molecular analyses were used to confirm diagnosis at species level. In addition to few existing reports of human infection by O. lupi in Turkey, this case further suggests that the pathogen might be more common than previously reported, thus requiring further attention and investigations.
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- 2021
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37. Detection of Leishmania tarentolae in lizards, sand flies and dogs in southern Italy, where Leishmania infantum is endemic: hindrances and opportunities.
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Mendoza-Roldan JA, Latrofa MS, Iatta R, R S Manoj R, Panarese R, Annoscia G, Pombi M, Zatelli A, Beugnet F, and Otranto D
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- Animals, Dogs, Female, Leishmania classification, Leishmania genetics, Leishmania immunology, Leishmania infantum pathogenicity, Male, Zoonoses epidemiology, Zoonoses parasitology, Zoonoses transmission, Endemic Diseases veterinary, Leishmania isolation & purification, Leishmania infantum isolation & purification, Leishmaniasis epidemiology, Leishmaniasis veterinary, Lizards parasitology, Psychodidae parasitology
- Abstract
Background: Leishmania tarentolae is a protozoan isolated from geckoes (Tarentola annularis, Tarentola mauritanica), which is considered non-pathogenic and is transmitted by herpetophilic Sergentomyia spp. sand flies. This species occurs in sympatry with Leishmania infantum in areas where canine leishmaniasis is endemic. In the present study, we investigated the circulation of L. tarentolae and L. infantum in sand flies, dogs and lizards in a dog shelter in southern Italy, where canine leishmaniasis by L. infantum is endemic., Methods: Sheltered dogs (n = 100) negative for Leishmania spp. (March 2020) were screened by immunofluorescence antibody test (IFAT) using promastigotes of both species at two time points (June 2020 and March 2021). Whole blood from dogs, tissues of Podarcis siculus lizards (n = 28) and sand flies (n = 2306) were also sampled and tested by a duplex real-time PCR (dqPCR). Host blood meal was assessed in sand flies by PCR., Results: Overall, 16 dogs became positive for L. infantum and/or L. tarentolae by IFAT at one or both sampling periods. One canine blood sample was positive for L. infantum, whilst two for L. tarentolae by dqPCR. At the cytology of lizard blood, Leishmania spp. amastigote-like forms were detected in erythrocytes. Twenty-two tissue samples, mostly lung (21.4%), scored molecularly positive for L. tarentolae, corresponding to 10 lizards (i.e., 35.7%). Of the female Sergentomyia minuta sampled (n = 1252), 158 scored positive for L. tarentolae, four for L. infantum, and one co-infected. Two Phlebotomus perniciosus (out of 29 females) were positive for L. tarentolae. Engorged S. minuta (n = 10) fed on humans, and one P. perniciosus, positive for L. tarentolae, on lagomorphs., Conclusions: Dogs and lacertid lizards (Podarcis siculus) were herein found for the first time infected by L. tarentolae. The detection of both L. tarentolae and L. infantum in S. minuta and P. perniciosus suggests their sympatric circulation, with a potential overlap in vertebrate hosts. The interactions between L. tarentolae and L. infantum should be further investigated in both vectors and vertebrate hosts to understand the potential implications for the diagnosis and control of canine leishmaniasis in endemic areas., (© 2021. The Author(s).)
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- 2021
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38. Marked host association and molecular evidence of limited transmission of ticks and fleas between sympatric wild foxes and rural dogs.
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Cevidanes A, Ulloa-Contreras C, Di Cataldo S, Latrofa MS, Gonzalez-Acuña D, Otranto D, and Millán J
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- Animals, Dogs, Foxes, Ctenocephalides, Dog Diseases epidemiology, Flea Infestations epidemiology, Flea Infestations veterinary, Siphonaptera, Ticks
- Abstract
Wild and domestic carnivores share ectoparasites, although molecular evidence is lacking. The goals of this study were to describe tick and flea infestation in sympatric free-ranging dogs Canis lupus familiaris (Linnaeus, 1758) (Carnivora: Canidae) and Andean foxes Lycalopex culpaeus (Molina, 1782) (Carnivora: Canidae) and to determine whether interspecific transmission occurs. Fleas and ticks retrieved from 79 foxes and 111 dogs in the human-dominated landscapes of central Chile were identified and a subset of specimens characterized by PCR and amplicon sequencing. Each ectoparasite species was clearly associated with a host: abundance and occurrence of Rhipicephalus sanguineus (Latreille 1806) (Acari: Ixodidae) and Ctenocephalides spp. (Siphonaptera: Pulicidae) were significantly higher in dogs than in foxes, whereas the opposite was true for Amblyomma tigrinum (Koch, 1844) (Acari: Ixodidae) and Pulex irritans (Linnaeus, 1758) (Siphonaptera: Pulicidae). Genetic analyses of a subset of ectoparasites revealed that dogs and foxes shared a limited number of nucleotide sequence types, suggesting that the interspecific transmission of these ectoparasites happens infrequently. Data also indicated that the ecological association and biological cycles of ticks and fleas determine the ectoparasite fauna of sympatric carnivores. In conclusion, our study shows that cross-species transmission should be assessed at a molecular level., (© 2021 The Royal Entomological Society.)
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- 2021
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39. Molecular detection of zoonotic filarioids in Culex spp. from Portugal.
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Manoj RRS, Latrofa MS, Cavalera MA, Mendoza-Roldan JA, Maia C, and Otranto D
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- Animals, Mosquito Vectors, Portugal, Aedes, Culex, Dirofilaria immitis
- Abstract
To investigate the role of dipterans in the transmission of Onchocerca lupi and other zoonotic filarioids, samples were collected from different sites in Algarve, southern Portugal, morphologically identified and molecularly tested for filarioids. Culex sp. (72.8%) represented the predominant genus followed by Culicoides sp. (11.8%), Ochlerotatus sp. (9.7%), Culiseta sp. (4.5%), Aedes sp. (0.9%) and Anopheles sp. (0.3%). Nineteen (2.8%) specimens scored positive for filarioids, with Culex pipiens quinquefasciatus (2%) positive for Dirofilaria immitis (1.4%), Dirofilaria repens, Acanthocheilonema reconditum, Onchocerca lupi, unidentified species of Filarioidea (0.2%, each) and Onchocercidae (0.6%). Additionally, Culiseta longiareolata (6.5%), Ochlerotatus caspius (3%) and Culex laticinctus (0.2%) scored positive for unidentified Onchocercidae, A. reconditum and for O. lupi, respectively. This is the first report of the occurrence of DNA of O. lupi, D. repens and A. reconditum in Culex spp. in Portugal. Information regarding the vectors and the pathogens they transmit may help to adopt proper prophylactic and control measures., (© 2021 The Authors. Medical and Veterinary Entomology published by John Wiley & Sons Ltd on behalf of The Royal Entomological Society.)
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- 2021
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40. Molecular detection of Wolbachia endosymbiont in reptiles and their ectoparasites.
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Manoj RRS, Latrofa MS, Mendoza-Roldan JA, and Otranto D
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- Animals, Phylogeny, RNA, Ribosomal, 16S genetics, Reptiles parasitology, Ixodes microbiology, Mites microbiology, Reptiles microbiology, Wolbachia genetics, Wolbachia isolation & purification
- Abstract
Wolbachia, a maternally transmitted Gram-negative endosymbiont of onchocercid nematodes and arthropods, has a role in the biology of their host; thus it has been exploited for the filariasis treatment in humans. To assess the presence and prevalence of this endosymbiont in reptiles and their ectoparasites, blood and tail tissue as well as ticks and mites collected from them were molecularly screened for Wolbachia DNA using two sets of primers targeting partial 16S rRNA and Wolbachia surface protein (wsp) genes. Positive samples were screened for the partial 12S rRNA and cytochrome c oxidase subunit 1 (cox1) genes for filarioids. Of the different species of lizards (Podarcis siculus, Podarcis muralis and Lacerta bilineata) and snakes (Elaphe quatuorlineata and Boa constrictor constrictor) screened from three collection sites, only P. siculus scored positive for Wolbachia 16S rRNA. Among ectoparasites collected from reptiles (Ixodes ricinus ticks and Neotrombicula autumnalis, Ophionyssus sauracum and Ophionyssus natricis mites), I. ricinus (n = 4; 2.8%; 95% CI, 0.9-7) from P. siculus, N. autumnalis (n = 2 each; 2.8%; 95% CI, 0.9-6.5) from P. siculus and P. muralis and O. natricis (n = 1; 14.3%; 95% CI, 0.7-55.4) from Boa constrictor constrictor scored positive for Wolbachia DNA. None of the positive Wolbachia samples scored positive for filarioids. This represents the first report of Wolbachia in reptilian hosts and their ectoparasites, which follows a single identification in the intestinal cells of a filarioid associated with a gecko. This data could contribute to better understand the reptile filarioid-Wolbachia association and to unveil the evolutionary pattern of Wolbachia in its filarial host., (© 2021. The Author(s).)
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- 2021
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41. Fasciola hepatica in wild boar (Sus scrofa) from Italy.
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Sgroi G, Iatta R, Lia RP, Latrofa MS, Annoscia G, Veneziano V, and Otranto D
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- Animals, Brazil, Cattle, Egypt, Iran, Italy epidemiology, Phylogeny, Sheep, Sus scrofa, Swine, Cattle Diseases, Fasciola hepatica genetics, Goat Diseases, Sheep Diseases, Swine Diseases epidemiology
- Abstract
Fasciola hepatica is a trematode infecting ruminants worldwide, occasionally reported in a wide range of animal species, including humans. According to the WHO, fasciolosis is recognized as a re-emerging neglected tropical disease, responsible for endemic and epidemic outbreaks in humans. Although the main hosts of the parasite are represented by cattle, sheep and goats, wildlife may be involved in its circulation. Here we firstly report F. hepatica in a wild boar from Italy (southern area) and characterize it both morphologically and molecularly. The nad1 gene analysis of specimens analyzed, revealed a high genetic similarity with those of humans from Iran and Peru, as well as a close phylogenetic relationship to those in ruminants from Brazil, Ecuador and Egypt. Considering the increase in the wild boar populations in urban and peri-urban areas, a potential role of this ungulate in the circulation of this zoonotic trematode is suggested., (Copyright © 2021 Elsevier Ltd. All rights reserved.)
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- 2021
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42. Angiostrongylus vasorum in foxes ( Vulpes vulpes ) and wolves ( Canis lupus italicus) from Abruzzo region, Italy.
- Author
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Tieri EE, Saletti MA, D'Angelo AR, Parisciani G, Pelini S, Cocco A, Di Teodoro G, Di Censo E, D'Alterio N, Latrofa MS, Otranto D, and Pascucci I
- Abstract
In Europe wildlife animals such as the red fox ( Vulpes vulpes ) are considered the main reservoir for Angiostrongylus vasorum as well as a potential threat for domestic dog infection. Though this parasite is endemic in fox populations, data on A. vasorum infection in wolves ( Canis lupus italicus ) are still scant, having only recently been described in Northwestern Spain, in Italy, in Croatia and in Slovakia. Based on the rising number of cases of canine lungworm infection in Central Italy (Abruzzo region), the aim of the present study was to investigate the infection by A. vasorum in fox and wolf populations sharing the same geographical area of dogs. From October 2008 to November 2019, A. vasorum specimens were collected, through routine post-mortem examination, from 56 carcasses (44 foxes and 12 wolves). Adult parasites were searched for in the right side of the heart and in pulmonary artery of all carcasses. First stage of larvae (L1) was searched in faeces using the Baermann technique and in lungs by tissue impressions. Overall, 230 adult specimens were collected and identified on a morphological basis. To confirm the morphological identification, 4 adult specimens (n = 3 from fox, n = 1 from wolf) were molecularly identified as A. vasorum by amplification of partial fragment of nuclear 18S rRNA (~1700 bp) genes. The anatomo-pathological and parasitological examinations indicated the presence of A. vasorum in 33 foxes (75%) and in 8 wolves (66.7%). The level of prevalence of infested wolves was higher than the previous one reported in other European countries. Interestingly, the prevalence of infection in foxes herein recorded was higher than that described in dogs (8.9%) living in the same geographical area. This result may confirm the hypothesis that the spread of canine angiostrongylosis is linked to fox populations infection., Competing Interests: None., (© 2021 The Authors.)
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- 2021
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43. Wolbachia: endosymbiont of onchocercid nematodes and their vectors.
- Author
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Manoj RRS, Latrofa MS, Epis S, and Otranto D
- Subjects
- Animals, Arthropods microbiology, Arthropods physiology, Host-Pathogen Interactions, Nematoda growth & development, Nematoda immunology, Wolbachia genetics, Nematoda microbiology, Symbiosis, Wolbachia physiology
- Abstract
Background: Wolbachia is an obligate intracellular maternally transmitted, gram-negative bacterium which forms a spectrum of endosymbiotic relationships from parasitism to obligatory mutualism in a wide range of arthropods and onchocercid nematodes, respectively. In arthropods Wolbachia produces reproductive manipulations such as male killing, feminization, parthenogenesis and cytoplasmic incompatibility for its propagation and provides an additional fitness benefit for the host to protect against pathogens, whilst in onchocercid nematodes, apart from the mutual metabolic dependence, this bacterium is involved in moulting, embryogenesis, growth and survival of the host., Methods: This review details the molecular data of Wolbachia and its effect on host biology, immunity, ecology and evolution, reproduction, endosymbiont-based treatment and control strategies exploited for filariasis. Relevant peer-reviewed scientic papers available in various authenticated scientific data bases were considered while writing the review., Conclusions: The information presented provides an overview on Wolbachia biology and its use in the control and/or treatment of vectors, onchocercid nematodes and viral diseases of medical and veterinary importance. This offers the development of new approaches for the control of a variety of vector-borne diseases.
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- 2021
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44. Genetic variability of Ehrlichia canis TRP36 in ticks, dogs, and red foxes from Eurasia.
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Bezerra-Santos MA, Nguyen VL, Iatta R, Manoj RRS, Latrofa MS, Hodžić A, Dantas-Torres F, Mendoza-Roldan JA, and Otranto D
- Subjects
- Animals, Asia epidemiology, Bacterial Proteins genetics, Dogs, Ehrlichia canis classification, Ehrlichiosis epidemiology, Ehrlichiosis microbiology, Ehrlichiosis veterinary, Europe epidemiology, Gene Expression Regulation, Bacterial, Global Health, Phylogeny, RNA, Bacterial genetics, RNA, Ribosomal, 16S genetics, Bacterial Proteins metabolism, Dog Diseases microbiology, Ehrlichia canis genetics, Foxes microbiology, Genetic Variation, Rhipicephalus sanguineus microbiology
- Abstract
Ehrlichia canis is among the most prevalent tick-borne pathogens infecting dogs worldwide, being primarily vectored by brown dog ticks, Rhipicephalus sanguineus sensu lato (s.l.). The genetic variability of E. canis has been assessed by analysis of different genes (e.g., disulfide bond formation protein gene, glycoprotein 19, tandem repeat protein 36 - TRP36) in the Americas, Africa, Asia, and in a single dog sample from Europe (i.e., Spain). This study was aimed to assess the variations in the TRP36 gene of E. canis detected in naturally infected canids and R. sanguineus s.l. ticks from different countries in Asia and Europe. DNA samples from dogs (n = 644), foxes (n = 146), and R. sanguineus s.l. ticks (n = 658) from Austria, Italy, Iran, Pakistan, India, Indonesia, Malaysia, the Philippines, Singapore, Thailand, Vietnam, and Taiwan were included in this study. Ehrlichia canis 16S rRNA positive samples (n = 115 from the previous studies; n = 14 from Austria in this study) were selected for molecular examination by analyses of TRP36 gene. Out of 129 E. canis 16S rRNA positive samples from dogs (n = 88), foxes (n = 7), and R. sanguineus s.l. ticks (n = 34), the TRP36 gene was successfully amplified from 52. The phylogenetic analysis of the TRP36 pre-repeat, tandem repeat, and post repeat regions showed that most samples were genetically close to the United States genogroup, whereas two samples from Austria and one from Pakistan clustered within the Taiwan genogroup. TRP36 sequences from all samples presented a high conserved nucleotide sequence in the tandem repeat region (from 6 to 20 copies), encoding for nine amino acids (i.e., TEDSVSAPA). Our results confirm the US genogroup as the most frequent group in dogs and ticks tested herein, whereas the Taiwan genogroup was present in a lower frequency. Besides, this study described for the first time the US genogroup in red foxes, thus revealing that these canids share identical strains with domestic dogs and R. sanguineus s.l. ticks., (Copyright © 2021 Elsevier B.V. All rights reserved.)
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- 2021
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45. Role of reptiles and associated arthropods in the epidemiology of rickettsioses: A one health paradigm.
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Mendoza-Roldan JA, Ravindran Santhakumari Manoj R, Latrofa MS, Iatta R, Annoscia G, Lovreglio P, Stufano A, Dantas-Torres F, Davoust B, Laidoudi Y, Mediannikov O, and Otranto D
- Subjects
- Adult, Aged, Anaplasma phagocytophilum, Anaplasmataceae genetics, Animals, Coxiella burnetii, Europe, Female, Humans, Italy, Ixodes microbiology, Male, Middle Aged, Polymerase Chain Reaction, RNA, Ribosomal, 16S, Rickettsia, Snakes, Young Adult, Arthropods microbiology, One Health, Reptiles microbiology, Rickettsia Infections epidemiology, Rickettsia Infections veterinary
- Abstract
We assessed the presence of Rickettsia spp., Coxiella burnetii and Anaplasma phagocytophilum in reptiles, their ectoparasites and in questing ticks collected in a nature preserve park in southern Italy, as well as in a peri-urban area in another region. We also investigated the exposure to these pathogens in forestry workers, farmers and livestock breeders living or working in the nature preserve park given the report of anecdotal cases of spotted fever rickettsioses. Rickettsia spp. were molecularly detected in Podarcis muralis and Podarcis siculus lizards (i.e., 3.1%), in Ixodes ricinus (up to 87.5%) and in Neotrombicula autumnalis (up to 8.3%) collected from them as well as in I. ricinus collected from the environment (up to 28.4%). Rickettsia monacensis was the most prevalent species followed by Rickettsia helvetica. An undescribed member of the family Anaplasmataceae was detected in 2.4% and 0.8% of the reptiles and ectoparasites, respectively. Sera from human subjects (n = 50) were serologically screened and antibodies to Rickettsia spp. (n = 4; 8%), C. burnetti (n = 8; 16%) and A. phagocytophilum (n = 11; 22%) were detected. Two ticks collected from two forestry workers were positive for spotted fever group (SFG) rickettsiae. Ixodes ricinus is involved in the transmission of SFG rickettsiae (R. monacensis and R. helvetica) in southern Europe and lizards could play a role in the sylvatic cycle of R. monacensis, as amplifying hosts. Meanwhile, N. autumnalis could be involved in the enzootic cycle of some SFG rickettsiae among these animals. People living or working in the southern Italian nature preserve park investigated are exposed to SFG rickettsiae, C. burnetii and A. phagocytophilum., Competing Interests: The authors have declared that no competing interests exist.
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- 2021
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46. Major antigen and paramyosin proteins as candidate biomarkers for serodiagnosis of canine infection by zoonotic Onchocerca lupi.
- Author
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Latrofa MS, Palmisano G, Annoscia G, Pierri CL, Chandrashekar R, and Otranto D
- Subjects
- Animals, Biomarkers blood, Dog Diseases parasitology, Dogs, Female, Male, Microfilariae genetics, Microfilariae isolation & purification, Onchocerca immunology, Onchocerca isolation & purification, Onchocerciasis immunology, Onchocerciasis parasitology, Onchocerciasis, Ocular blood, Onchocerciasis, Ocular immunology, Onchocerciasis, Ocular parasitology, Serologic Tests, Tropomyosin blood, Tropomyosin isolation & purification, Dog Diseases diagnosis, Onchocerca chemistry, Onchocerciasis diagnosis, Onchocerciasis, Ocular diagnosis, Tropomyosin genetics, Tropomyosin immunology
- Abstract
Onchocerca lupi (Spirurida: Onchocercidae) is a filarial worm parasitizing domestic carnivores and humans. Adult nematodes usually localize beneath in the sclera or in the ocular retrobulbar of infected animals, whilst microfilariae are found in the skin. Therefore, diagnosis of O. lupi is achieved by microscopic and/or molecular detection of microfilariae from skin biopsy and/or surgical removal of adults from ocular tissues of infected hosts. An urgent non-invasive diagnostic tool for the diagnosis of O. lupi in dog is mandatory. In this study, an immunoproteomic analyses was performed using a combination of immunoblotting and mass spectrometry techniques. Onchocerca lupi major antigen (Ol-MJA) and paramyosin (Ol-PARA) proteins were identified as potential biomarkers for serodiagnosis. Linear epitopes were herein scanned for both proteins using high-density peptide microarray. Sera collected from dog infected with O. lupi and healthy animal controls led to the identification of 11 immunodominant antigenic peptides (n = 7 for Ol-MJA; n = 4 for Ol-PARA). These peptides were validated using sera of dogs uniquely infected with the most important filarioids infesting dogs either zoonotic (Dirofilaria repens, Dirofilaria immitis) or not (Acanthocheilonema reconditum and Cercopithifilaria bainae). Overall, six antigenic peptides, three for Ol-MJA and for Ol-PARA, respectively, were selected as potential antigens for the serological detection of canine O. lupi infection. The molecular and proteomic dataset herein reported should provide a useful resource for studies on O. lupi toward supporting the development of new interventions (drugs, vaccines and diagnostics) against canine onchocercosis., Competing Interests: We have read the journal's policy and the authors of this manuscript have the following competing interests: patent application filed and jointly submitted by the institution to which the following authors LMS, OD and PG are affiliated and/or from which the authors may benefit. RC is employee of IDEXX Laboratories, which funded partially the study. AG and PCL have no competing interests. IDEXX played a role in the review, approval and decision to publish of the manuscript.
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- 2021
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47. Molecular detection of vector-borne agents in ectoparasites and reptiles from Brazil.
- Author
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Mendoza-Roldan JA, Ribeiro SR, Castilho-Onofrio V, Marcili A, Simonato BB, Latrofa MS, Benelli G, Otranto D, and Barros-Battesti DM
- Subjects
- Animals, Brazil, Disease Reservoirs, Eucoccidiida classification, Female, Iridoviridae classification, Larva growth & development, Larva microbiology, Larva parasitology, Larva virology, Male, Nymph growth & development, Nymph microbiology, Nymph parasitology, Nymph virology, Phylogeny, Rickettsia classification, Eucoccidiida isolation & purification, Iridoviridae isolation & purification, Ixodidae growth & development, Ixodidae microbiology, Ixodidae parasitology, Ixodidae virology, Mites growth & development, Mites microbiology, Mites parasitology, Mites virology, Reptiles microbiology, Reptiles parasitology, Reptiles virology, Rickettsia isolation & purification
- Abstract
Trombidiformes and Mesostigmata mites, as well as Ixodida ticks, infest ectothermic tetrapods worldwide, potentially acting as vectors of bacteria, viruses and protozoa. The relationship among ectoparasites, transmitted pathogenic agents (e.g., Borrelia spp., Coxiella spp., Hepatozoon spp., and Rickettsia spp.) and ectothermic hosts has been scarcely investigated. This research focuses on a large collection of Brazilian herpetofauna screened for the presence of arthropod ectoparasites and vector-borne microbial agents. Reptiles (n = 121) and amphibians (n = 49) from various locations were infested by ectoparasites. Following genomic extraction, microbial agents were detected in 81 % of the Acari (i.e. n = 113 mites and n = 26 ticks). None of the mites, ticks and tissues from amphibians yielded positive results for any of the screened agents. Blood was collected from reptiles and processed through blood cytology and molecular analyses (n = 48). Of those, six snakes (12.5 %) showed intraerythrocytic alterations compatible with Hepatozoon spp. gamonts and Iridovirus inclusions. Hepatozoon spp. similar to Hepatozoon ayorgbor and Hepatozoon musa were molecularly identified from seven hosts, two mite and two tick species. Rickettsia spp. (e.g., Rickettsia amblyommatis, Rickettsia bellii-like, Rickettsia sp.) were detected molecularly from four mite species and Amblyomma rotundatum ticks. Phylogenetic analyses confirmed the molecular identification of the above-mentioned microbial agents of mites and ticks related to snakes and lizards. Overall, our findings highlighted that the Brazilian herpetofauna and its ectoparasites harbour potentially pathogenic agents, particularly from the northern and south-eastern regions. The detection of several species of spotted fever group Rickettsia pointed out the potential role of ectothermic hosts and related arthropod ectoparasites in the epidemiological cycle of these bacteria in Brazil., (Copyright © 2020 Elsevier GmbH. All rights reserved.)
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- 2021
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48. Molecular Approach for the Diagnosis of Blood and Skin Canine Filarioids.
- Author
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Laidoudi Y, Bedjaoui S, Medkour H, Latrofa MS, Mekroud A, Bitam I, Davoust B, Otranto D, and Mediannikov O
- Abstract
The zoonotic Onchocerca lupi and tick-transmitted filarioids of the genus Cercopithifilaria remain less well known due to the difficulties in accessing to skin samples as target tissues. Here, we proposed a molecular approach reliying on multiplex qPCR assays that allow the rapid identification of filarioids from canine blood, skin, and tick samples. This includes two newly developed duplex qPCR tests, the first one targeting filarial and C. grassii DNA (CanFil- C. grassii ). and the second qPCR assay designed for the detection of Cercopithifilaria bainae and Cercopithifilaria sp. II DNAs ( C. bainae -C.spII). The third one is a triplex TaqMan cox 1 assay targeting DNA of blood microfilariae (e.g., Dirofilaria immitis, Dirofilaria repens and Acanthocheilonema reconditum ). The novel duplex qPCRs developed were validated in silico and by screening of known DNA collection. The qPCR assays were also used for screening the blood and tick samples of 72 dogs from Algeria. This allowed the identification of canine filariasis infection with 100% of specificity and 89.47% and 100% of sensitivity from naturally infected blood and tick samples, respectively. The prevalences of 26.39% for D. immitis and 5.56% for both D. repens and A. reconditum were reported in blood and tick samples. Cercopithifilaria DNAs were detected only in tick samples, with a prevalence of 4.17% and 5.56% for C. bainae and Cercopithifilaria sp. II, respectively. Co-infections were diagnosed in 6.94% and 13.89% of blood and tick samples, respectively. Whereas all samples were negative for C. grassii DNA. The use of engorged ticks instead of blood and skin samples could be an easier option for the surveillance of all canine filarioids herein investigated. The multiplex qPCR assays herein validated were shown to be useful in the detection of filarial co-infections by overcoming sequencing of positive samples.
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- 2020
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49. Cercopithifilaria species in dogs and ticks from Greece.
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Angelou A, Latrofa MS, Annoscia G, Symeonidou I, Theodoridis A, Polizopoulou ZS, Otranto D, and Papadopoulos E
- Subjects
- Animals, Dog Diseases epidemiology, Dogs, Filariasis epidemiology, Filariasis parasitology, Filariasis transmission, Filarioidea classification, Filarioidea genetics, Genetic Variation, Greece epidemiology, Haplotypes, Arachnid Vectors parasitology, Dog Diseases parasitology, Filariasis veterinary, Filarioidea isolation & purification, Ticks parasitology
- Abstract
Filarioids of the genus Cercopithifilaria (Spirurida, Onchocercidae) are parasites of wild and domestic animals in tropical and subtropical regions being transmitted by ixodid ticks. Though this filarioid species have been studied in canine and tick populations in Europe, data on their species diversity and geographical distribution in Greece is scant. Thus, the aims of this study were to investigate the presence of Cercopithifilaria spp. in dogs and ticks across Greece and to assess the possible risk factors. A total of 500 skin biopsies were collected from dogs, while 508 ticks were collected from 180 infested animals and examined. Sediments from skin biopsies were microscopically screened for detection of dermal microfilaria (mfs). Skin samples (n = 115) and tick specimens (n = 153) were molecularly subjected by PCR. Overall, 70 samples (14%) scored positive for mfs. Specifically, 68 samples (13.6%) were positive for Cercopithifilaria bainae and two (0.4%) were co-infected with C. bainae and Cercopithifilaria sp. II. Molecular analyses revealed that all sequences obtained belong to C. bainae. Haplotype I was the most frequent (92.6%), followed by haplotype XVIII (3%) and haplotypes II and IX (1.5%). Three new haplotypes of C. bainae, named XIX, XX, and XXI, were also identified. Among the risk factors examined, habitat, dog use, body weight, tick infestation history, and the use of acaricides were associated with the presence of C. bainae. The estimated prevalence of Cercopithifilaria spp. demonstrates that these filarioids are common in dogs and ticks in Greece. Finally, the identification of 7 haplotypes for C. bainae confirms their genetic variability.
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- 2020
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50. Hyperendemic Dirofilaria immitis infection in a sheltered dog population: an expanding threat in the Mediterranean region.
- Author
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Panarese R, Iatta R, Latrofa MS, Zatelli A, Ćupina AI, Montarsi F, Pombi M, Mendoza-Roldan JA, Beugnet F, and Otranto D
- Subjects
- Animals, Dogs parasitology, Endemic Diseases veterinary, Italy epidemiology, Mediterranean Region epidemiology, Mosquito Vectors, Prevalence, Dirofilaria immitis, Dirofilariasis epidemiology, Dog Diseases parasitology, Dog Diseases transmission
- Abstract
A study on the occurrence of Dirofilaria immitis and its vectors was carried out in order to assess the prevalence of the disease in dogs in previously non-endemic areas of southern Italy. Blood samples (n = 385) and mosquitoes (n = 1540) were collected in two dog shelters and analysed by Knott's test and duplex real-time PCR, respectively. Dirofilaria immitis was the most prevalent filarioid (44.2%), while Culex pipiens was the most prevalent mosquito species (68.8%). This high prevalence of D. immitis infection confirms this location as one of the most hyperendemic foci of dirofilariosis in Europe., (Copyright © 2020 Australian Society for Parasitology. Published by Elsevier Ltd. All rights reserved.)
- Published
- 2020
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