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Soares Pereira da Fonseca de Sampaio, Isabel Maria
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- Meglumine antimoniate and miltefosine combined with allopurinol sustain pro-inflammatory immune environments during canine leishmaniosis treatmentPublication . Santos, Marcos André Ferreira; Alexandre-Pires, Graça; Pereira, Maria A.; Marques, Cátia Filipa Saraiva; Gomes, Joana; Correia, Jorge Manuel de Jesus; Duarte, Ana; Gomes, Lídia; Rodrigues, Armanda; Basso, Alexandra; Reisinho, Ana; Meireles, José; Santos-Mateus, David; Villa Brito, Maria Teresa; Tavares, Luis; Santos-Gomes, Gabriela M.; Pereira da Fonseca, IsabelCanine leishmaniosis (CanL) caused by Leishmania infantum is a zoonotic disease of global concern. Antileishmanial drug therapies commonly used to treat sick dogs improve their clinical condition, although when discontinued relapses can occur. Thus, the current study aims to evaluate the effect of CanL treatments in peripheral blood, lymph node, and bone marrow cytokine profile associated with clinical recovery. Two groups of six dogs diagnosed with CanL were treated with miltefosine combined with allopurinol and meglumine antimoniate combined with allopurinol (MT+A and MG+A), respectively. At diagnosis and after treatment, during a 3-month follow-up, clinical signs, hematological and biochemical parameters, urinalysis results and antileishmanial antibody titers were registered. Furthermore, peripheral blood, popliteal lymph node, and bone marrow samples were collected to assess the gene expression of IL-2, IL-4, IL-5, IL-10, IL-12, TNF-alpha, TGF-beta, and IFN-gamma by qPCR. In parallel, were also evaluated samples obtained from five healthy dogs. Both treatment protocols promoted the remission of clinical signs as well as normalization of hematological and biochemical parameters and urinalysis values. Antileishmanial antibodies returned to non-significant titers in all dogs. Sick dogs showed a generalized upregulation of IFN-gamma and downregulation of IL-2, IL-4, and TGF-beta, while gene expression of IL-12, TNF-alpha, IL-5, and IL-10 varied between groups and according to evaluated tissue. A trend to the normalization of cytokine gene expression was induced by both miltefosine and meglumine antimoniate combined therapies. However, IFN-gamma gene expression was still up-regulated in the three evaluated tissues. Furthermore, the effect of treatment in the gene expression of cytokines that were not significantly changed by infection, indicates that miltefosine and meglumine antimoniate combined therapy directly affects cytokine generation. Both combined therapies are effective in CanL treatment, leading to sustained pro-inflammatory immune environments that can compromise parasite survival and favor dogs' clinical cure. In the current study, anti-inflammatory and regulatory cytokines do not seem to play a prominent role in CanL or during clinical recovery.
- Diagnosis and clinical management of canine leishmaniosis by general veterinary practitioners: a questionnaire-based survey in PortugalPublication . Monteiro, Marta; Prata, Sara; Cardoso, Luís; Pereira da Fonseca, Isabel; Leal, RodolfoBackground: Canine leishmaniosis (CanL) can be appropriately managed following international recommendations. However, few studies have assessed the preferred protocols in real-life veterinary practice and whether these are in line with the guidelines. This survey aimed to investigate the current trends in the clinical management of CanL among veterinary practitioners in Portugal, taking into consideration diferent scenarios of infection/disease and the awareness of and application by veterinary practitioners of the current guidelines. Methods: A questionnaire-based survey was conducted online using an electronic platform. The following topics were surveyed: (i) general characteristics of the responding veterinarian; (ii) the preferred protocols used for the diagnosis, treatment and prevention of CanL, considering diferent theoretical scenarios of infection/disease; and (iii) the responding veterinarian’s current knowledge and application of the existing guidelines on CanL. After internal validation, the survey was distributed online, for 2 months, via Portuguese social network veterinary groups. Data were collected for descriptive analysis. Results: Eighty-six replies were obtained. Analysis of the results showed that the preferred diagnostic techniques varied widely according to the theoretical scenario of infection/disease. In general daily practice, serology testing (enzyme-linked immunosorbent assay [ELISA]) was the most used tool (67.4%). The preferred matrices used for PCR test were lymph nodes (62.3%) and/or bone marrow (59.0%). Regarding treatment, for subclinical infection/stage I CanL, 51.2% of the respondents did not prescribe any medical treatment, but 98.8% proceeded with both monitoring and preventive measures. Among those who prescribed a treatment (n=42), most chose domperidone (47.6%). For the treament of stages IIa, IIb and III CanL, allopurinol/meglumine antimoniate (MA) was chosen by 69.8, 73.3 and 51.2% of respondents, respectively, followed by allopurinol/miltefosine (20.9, 19.8 and 38.4%, respectively). In contrast, dogs with stage IV CanL were mostly treated with allopurinol/miltefosine (48.8%) rather than with allopurinol/MA (23.3%). The use of repellents was the preferred preventive strategy (98.8%). About 93.0% of responders were aware of the existence of guidelines, and most of these veterinarians consulted the guidelines of the LeishVet group and the Canine Leishmaniosis Working Group; however, 31.3% reported that they did not follow any specifc recommendations.
- Coccidian parasites from birds at rehabilitation centers in Portugal, with notes on Avispora bubonis in Old WorldPublication . Cardozo, Sergian Vianna; Berto, Bruno Pereira; Caetano, Inês; Thomas, André; Santos, Marcos; Pereira da Fonseca, Isabel; Gomes Lopes, Carlos WilsonABSTRACT - Portugal has some rehabilitation centers for wild animals, which are responsible for the rehabilitation and reintroduction of birds, among other animals, into the wild. Coccidian parasites of these wild birds in rehabilitation centers are especially important because these centers can introduce coccidian species into new environments through the reintroduction of their respective hosts. In this context, the current study aimed to identify intestinal coccidia from wild birds at two rehabilitation centers for wild animals located in two municipalities of Portugal. Eighty-nine wild birds of 9 orders and 11 families were sampled, of which 22 (25%) were positive for Coccidia. Avispora spp. were found in raptors. Sporocysts of Sarcocystinae subfamily were recovered from owls. An Isospora sp. was found in Turdus merula Linnaeus, 1758, and an Eimeria sp. was found in Fulica atra Linnaeus, 1758. Among the coccidian species, Avispora bubonis (Cawthorn, Stockdale, 1981) can be highlighted. The finding of this species indicates that transmission of coccidians from the New World to the Old World may be occurring, potentially through dispersion by Bubo scandiacus (Linnaeus, 1758) through Arctic regions or by means of anthropic activities, and/or through other unknown mechanisms.
- Dogs with leishmaniosis: how are we managing proteinuria in daily practice? A Portuguese questionnaire-based studyPublication . Monteiro, Marta; Prata, Sara; Cardoso, Luís; Pereira da Fonseca, Isabel; Leal, RodolfoABSTRACT Background: Proteinuria is a common fnding in dogs with leishmaniosis. Although antileishmanial therapeutic protocols are widely implemented, little information is available on which treatments are most adequate for identifying proteinuria in patients with canine leishmaniosis (CanL), especially regarding the use of immunosuppressants. The aim of this study was to explore the current paradigm regarding the antiproteinuric approach adopted by veterinary practitioners in Portugal to treat dogs with CanL. Methods: A questionnaire-based study was developed using Google Forms®. The questionnaire presented a number of diferent hypothetical scenarios of CanL, and the topics surveyed included the general features of the respondents and the protocols preferred by these respondents to manage proteinuria in the presented scenarios, including choice of therapeutic drugs, appropriate diet and use of immunosuppressants, in dogs with immune-mediated glomerulonephritis. The questionnaire was internally prevalidated and difused online over a 2-month period through Portuguese veterinary social networking groups, and data were collected for descriptive analysis. Results: A total of 86 veterinary practitioners responded to the survey. When exposed to theoretical scenarios of proteinuria in dogs with CanL at stages IIb, III and IV (LeishVet guidelines), 16.3%, 62.8% and 93.8% of the respondents, respectively, answered that they would treat it. The dog was started on a renal diet as therapy by 28.6%, 83.3% and 97.4% of respondents, respectively. Angiotensin-converting enzyme inhibitors (ACEI) were prescribed by 100%, 85.2% and 78.9% of respondents as frst-choice drugs for CanL at stages IIb, III and IV, respectively, with ACEI used in monotherapy by 64.3%, 40.7% and 46.1%. In comparison, protocols using ACEI in combination with other compounds were chosen by 7.1%, 33.3% and 39.5% of respondents, and combination therapy which did not include ACEI was the choice of 0.0%, 12.9% and 14.5%. Regarding immunosuppressants, 44.2% of the respondents answered they would prescribe them, with 97.4% electing for prednisolone and 5.3% choosing mycophenolate mofetil. Conclusions: Among the veterinary practitioners who responded, proteinuria treatment was considered since stage IIb CanL, although implementation of a therapeutic approach was more evident in advanced CanL stages. ACEI were the frst-choice drugs, particularly for the treatment of stage IIb CanL; in advanced stages, a combination of antiproteinuric drugs was more often used. Immunosuppressant use was controversial, although when applied, prednisolone was the preferred choice. These fndings reinforce the small body of evidence that supports the use of such drugs and the need to further explore their role in CanL.
- Zoonotic visceral leishmaniasis : new insights on innate immune response by blood macrophages and liver kupffer cells to leishmania infantum parasitesPublication . Rodrigues, Armanda Viana; Valerio-Bolas, Ana; Alexandre-Pires, Graça; Aires Pereira, Maria; Nunes, Telmo; Ligeiro, Dario; Pereira da Fonseca, Isabel; Santos-Gomes, GabrielaL. infantum is the aetiological agent of zoonotic visceral leishmaniasis (ZVL), a disease that affects humans and dogs. Leishmania parasites are well adapted to aggressive conditions inside the phagolysosome and can control the immune activation of macrophages (MØs). Although MØs are highly active phagocytic cells with the capacity to destroy pathogens, they additionally comprise the host cells for Leishmania infection, replication, and stable establishment in the mammal host. The present study compares, for the first time, the innate immune response to L. infantum infection of two different macrophage lineages: the blood macrophages and the liver macrophages (Kupffer cells, KC). Our findings showed that L. infantum takes advantage of the natural predisposition of blood-MØs to phagocyte pathogens. However, parasites rapidly subvert the mechanisms of MØs immune activation. On the other hand, KCs, which are primed for immune tolerance, are not extensively activated and can overcome the dormancy induced by the parasite, exhibiting a selection of immune mechanisms, such as extracellular trap formation. Altogether, KCs reveal a different pattern of response in contrast with blood-MØs when confronting L. infantum parasites. In addition, KCs response appears to be more efficient in managing parasite infection, thus contributing to the ability of the liver to naturally restrain Leishmania dissemination
- Presence of one ecto- and two endoparasite species of the black stork (Ciconia nigra) in PortugalPublication . Ramilo, David; Caetano, Inês; Brazio, Erica; Mira, Manuela; Antunes, Leonor; Pereira da Fonseca, Isabel; Cardoso, LuísBackgroundThe black stork (Ciconia nigra Linnaeus, 1758) is a recognized endangered species in Europe and most of the specimens from the Western Palearctic region breed in the Iberian Peninsula. Available works regarding parasites in black storks are scarce. This work reports the presence one ecto- and two endoparasite species from a black stork in Portugal.Case presentationA black stork was found in southern Portugal after colliding against electric cables. The specimen did not survive its sustained injuries and a post-mortem exam was performed. During the procedure, several ecto- and endoparasite specimens were found. The collected parasites were lice (Neophilopterus tricolor), nematodes (Desportesius sagittatus) and trematodes (Cathaemasia hians).ConclusionsThree different species of parasites are reported from a black stork in Portugal. Ecto- and endoparasites of C. nigra have not frequently been described in the literature, and this case report is a contribution to the field. Additional studies will be important to better understand the impact that parasites can have on C. nigra health and survival.
- Louse flies in Azorean and mainland populations of four Passeriformes species_a new perspective to parasite Island syndromesPublication . Tomás, André; Pereira da Fonseca, Isabel; Valkenburg, Thijs; Rebelo, Maria TeresaHippoboscid flies, also known as louse flies, are obligate blood-feeders ectoparasites of birds and mammals. By studying louse fly parasites of four Passeriformes species, Eurasian blackbird (Turdus merula), Eurasian blackcap (Sylvia atricapilla), common chaffinch (Fringilla coelebs) and European robin (Erithacus rubecula), with dissimilar time of colonization of Azores islands, we tested whether: (i) island host populations have lower parasite richness than the mainland one; (ii) island host populations undergo higher parasite prevalence, mean intensities and mean abundance than the mainland one; (iii) island parasite diversity are composed exclusively by specific parasites and (iv) parasite richness is positively correlated with the island area and proximity to the continent. For these purposes, 775 birds were sampled for presence of louse flies, by modified fumigation chamber method, from Azores Islands (S˜ao Miguel, Terceira and Flores) and Portugal mainland. Insular and mainland parasite assemblages were statistically compared. We record for the first time to Azores, Ornithomya fringillina and Icosta minor from Portugal mainland. Louse flies had highest prevalence and abundance from Azores Islands compared to those observed in mainland birds, especially blackbirds. The insular parasite diversity of Azores blackbirds, blackcaps and chaffinches was richer than the one observed in mainland population. None of the hippoboscid flies observed on the islands and mainland were host-specific. Thus, our findings provide an upgrade of parasite island syndromes knowledge, in the context of the ectoparasites, namely to the ippoboscid flies case.
- Molecular survey of vector-borne diseases in two groups of domestic dogs from Lisbon, PortugalPublication . Dordio, Ana Mafalda; Beck, Relja; Nunes, Telmo; Pereira da Fonseca, Isabel; Gomes, JacintoABSTRACT - Background: Canine vector-borne diseases (CVBDs) are caused by a wide range of pathogens transmitted by arthropods. They have been an issue of growing importance in recent years; however, there is limited information about the vector-borne pathogens circulating in Portugal. The aim of the present study was to detect canine vector-borne bacteria and protozoa of veterinary and zoonotic importance using molecular methods. Methods: One hundred and forty-two dogs from Lisbon, southern Portugal, were tested: 48 dogs from a veterinary hospital clinically suspected of vector-borne diseases and 94 apparently healthy dogs from shelters. Anaplasma spp./Ehrlichia spp., Babesia/Theileria spp., Hepatozoon spp., and Mycoplasma spp. infections were detected by PCR from blood samples and examined under light microscopy. Other information including clinical status and diagnostic test results were collected for each animal. Results: Infections were detected by PCR in 48 (33.80%) dogs. Single infections were found in 35 dogs (24.64%), and co-infections were found in 13 (9.15%) dogs. Twenty-nine (20.42%) dogs were positive for Hepatozoon spp., 15 (10.56%) for Mycoplasma spp., 11 (7.75%) for Anaplasma spp./Ehrlichia spp., and six (4.21%) for Babesia spp. DNA sequencing was used to identify Babesia vogeli (2.81%), Babesia canis (1.40%), Hepatozoon canis (20.42%), Mycoplasma haematoparvum (2.11%), Mycoplasma haemocanis (8.45%), Anaplasma platys (7.04%), and Ehrlichia canis (0.70%). Conclusions: This is the frst molecular identifcation of B. canis and M. haematoparvum in dogs from southern Portugal. This study highlights the importance of molecular methods to identify CVBD pathogens in endemic areas and helps to guide the clinical approach of veterinarians in practice.