Isabel Pereira da Fonseca

Survey of infectious and parasitic diseases in stray cats at the Lisbon Metropolitan Area, Portugal

A survey of infectious and parasitic diseases of stray cats was carried out using biological samples collected from animals captured during a catch-neuter-release programme in four counties of the Lisbon Metropolitan Area. The main objective was to investigate the potential threat of stray cats for animal and public health. Samples of blood, stool, hair and auricular swabs were collected from 231 cats in 27 colonies. Anti-Toxoplasma gondii antibodies were detected in 47/194 samples (24.2%); anti-Leishmania infantum antibodies in 1/180 cats (0.6%); intestinal parasites in 23/74 samples (Toxocara cati, Isospora felis, Ancylostoma tubaeforme, Dipylidium caninum, Uncinaria stenocephala, Toxascaris leonina) and Otodectes cynotis in 4/182 cats (2.2%); dermatophyte fungi were isolated in 40/136 samples (29.4%); feline immunodeficiency virus antibodies were detected in 23/226 samples (10.2%); feline leukaemia virus antigen in 14/198 samples (7.1%); and feline coronavirus RNA in 9/127 samples (7.1%). Our results revealed that zoonotic agents, namely dermatophyte fungi and Toxocara cati were present in stray cat colonies in the investigated counties. Overall the low frequency of major pathogens suggests a balanced relationship between host and agents.

MULTILOCUS GENOTYPING OF CRYPTOSPORIDIUM ISOLATES FROM HUMAN HIV-INFECTED AND ANIMAL HOSTS

’he genus Cryptosporidiwn comprises parasites of several species that infect mammals and other vertebrates. Molecular studies have drown that Cryptosporidiwn parvwn, the most common species infecting man, comprises the “human” genotype (H) found exclusively m humans and the “cattle” genotype (C) found in both humans and livestock [8]. Other genotypes, until now considered to be host specific, have also been found in mice, pigs, ferrets and marsupials [5, 141. Recently, FCR-RFLP and DNA sequence analysis have also identified C. felis, C. meleagridis and a Cryptosporidiwn “dog type” in HIVinfected patients as well as in immunmpetent persons [4, 6, 7. 9,

Canine leishmaniosis. Immunophenotypic profile of leukocytes in different compartments of symptomatic, asymptomatic and treated dogs

Canine visceral leishmaniasis (CanL) is an emerging disease, expanding in various parts of the world. The infection caused by Leishmania, an intracellular protozoan parasite, can show different clinical manifestations, from asymptomatic or subclinical to symptomatic dogs, in which a wide spectrum of clinical signs is evident. The fact that the parasite replicates in different organs raises the hypothesis that each organ may have a specific immune response. The local immune responses should be evaluated and taken into consideration when developing prophylactic tools. Therefore, phenotypic characterization of peripheral blood, lymph node and bone marrow lymphocyte populations and the expression of class II molecules of major histocompatibility complex (MHCII) were performed in asymptomatic and symptomatic dogs and in dogs that had been diagnosed and treated for leishmaniasis. Our findings showed that blood and bone marrow lymphocytes from symptomatic dogs were highly activated. In bone marrow of asymptomatic and treated dogs, a high frequency of MHCII(+) lymphocytes was observed, as well as MHCII(+) monocytes in the treated group. These results show increased expression of MHCII molecules giving evidence for antigenic presentation mainly by lymphocytes. The symptomatic and treated dogs showed an expansion of CD4(+) T cells subpopulations in lymph nodes, revealing an important contribution of these cells in controlling local parasite replication. This study also underlines the eventual importance of CD3(+)CD4(-)CD8(-) (double negative) and CD3(+)CD4(+)CD8(+) (double positive) T cell subsets in sensing and controlling latent infections and their possible function in the immune dynamics during CanL. The specific cellular immune responses raised in different compartments where the parasite replicates seem to have variable effects on local parasite control, highlighting the complexity of the cellular immune response developed by the dog infected by Leishmania infantum.

Natural infection of synathropic rodent species Mus musculus and Rattus norvegicus by Leishmania infantum in Sesimbra and Sintra – Portugal

BackgroundCanine leishmaniosis caused by Leishmania infantum is a parasitic zoonotic disease transmitted by phlebotomine sandflies (Diptera: Psychodidae). Genus Phlebotomus is the biological vector in the Old World and Lutzomyia in the New World. The dog is the domestic reservoir host but other animals like the fox (Vulpes vulpes) and rodents are known to maintain the infection in both sylvatic and domestic cycles.MethodsTo identify the role of synanthropic rodents Mus musculus and Rattus norvegicus as reservoir hosts for Leishmania infantum natural infection, 30 rodents were captured under a trap rodent control program in two private dog shelters from Sintra and Sesimbra, located in the Lisbon Metropolitan Area, known to be endemic for canine leishmaniosis in Portugal. Tissue samples were screened for the presence of Leishmania amastigotes by qPCR and parasitological analysis.ResultsA total of 33.3% (9/27) of Mus musculus rodents revealed the presence of Leishmania spp. DNA while 29.6% (8/27) were positive in the parasitological analysis. Concerning Rattus norvegicus (n=3), one animal revealed infection only by parasitological analysis.ConclusionsOur results identified for the first time in Portugal the presence of Leishmania infection in both rodent species. As susceptible hosts, infected Mus musculus and Rattus norvegicus may increase the risk for dog and human infection in households and surrounding areas, enhancing the need for efficient rodent control measures in shelters and risk zones to prevent transmission of the infection.

Hand length as an alternative measurement of height

Background/objectives:Despite the utmost importance of body height in evaluating nutritional status, it is not always possible to obtain its measurement and height may have to be estimated. The objective of the study was to formulate and cross-validate a regression equation to predict height using hand length measurement and also to determine if predicted height (PH) will lead to significant errors when used in body mass index (BMI) calculation.Subjects/methods:A cross-sectional study was conducted using a consecutive sample of 465 inpatients (19–91 years), from a university hospital. Participants were randomly divided into a development sample of 311 individuals and a cross-validation one. A linear regression model was used to formulate the equation. Intraclass correlation coefficients (ICCs) for single measures and differences between measured height (MH) and PH and between BMI calculated with MH (BMIMH) and with PH (BMIPH) were determined.Results:The regression equation for PH is: PH (cm)=80.400+5.122 × hand length (cm)—0.195 × age (years)+6.383 × gender (gender: women 0, men 1) (R=0.87, s.e. of the estimate=4.98 cm). MH and PH were strongly correlated, ICCs: 0.67-0.74 (P<0.001). Differences were small, mean difference±s.d., ⩽−0.6±4.4 cm (P⩾0.24). BMIMH and BMIPH were strongly correlated, ICCs: 0.94-0.96 (P<0.001). Differences were small, ⩽0.3±1.7 kg/m2 (P⩾0.10).Conclusions:The formulated regression equation using hand length, age and gender provides a valid estimation of height and is useful in the clinical context. PH from this regression equation can be used in BMI calculations as misclassification is small.

Financial impact of sarcopenia on hospitalization costs

Background/Objectives:This study aims to increase knowledge regarding the association of sarcopenia with hospitalization costs among a wide-ranging sample of adult hospitalized patients.Subjects/Methods:A prospective study was conducted among hospitalized adult patients. Sarcopenia was identified according to the European Working Group on Sarcopenia in Older People, as low muscle mass, assessed by bioelectrical impedance analysis and low muscle function evaluated by handgrip strength. Hospitalization cost was calculated for each patient based on discharge diagnosis-related group codes and determined on the basis of a relative weight value. Costs were defined as the percentage of deviation from the cost of a patient with a relative weight equal to one. Multivariable linear regression models were performed to identify the factors independently associated with hospitalization costs.Results:A total of 656 hospitalized patients aged ⩾18 years (24.2% sarcopenic) composed the study sample. Sarcopenia increased hospitalization costs by [euro ]1240 (95% confidence interval (CI): [euro ]596–1887) for patients aged <65 years and [euro ]721 (95% CI: [euro ]13–1429) for patients aged ⩾65 years. Sarcopenic overweight was related to an increase in hospitalization costs of [euro ]884 (95% CI: [euro ]295–1476).Conclusions:Sarcopenia is independently related to hospitalization costs. This condition is estimated to increase hospitalization costs by 58.5% for patients aged <65 years and 34% for patients aged ⩾65 years.

Helioid inclusions in dedifferentiated acinic cell carcinoma of the parotid gland

Helioid bodies are exceedingly rare, intranuclear, rounded inclusions with peripheral radiating filaments. These structures, which were formerly observed in only three cases of proliferative breast epithelial lesions, appeared in 0.5-1% of the neoplastic cells of a case of dedifferentiated acinic cell carcinoma of the parotid gland. Helioid bodies are related to rough endoplasmic reticulum cisternae, which suggests that they are the result of the condensation and partial crystallization of their contents when sequestrated into the nuclear compartment of the cell. The presence of helioid bodies may be related to secretory dysfunction of the tumor cells associated with the process of neoplastic dedifferentiation.

Management of canine leishmaniosis in endemic SW European regions: a questionnaire-based multinational survey

BackgroundCanine leishmaniosis (CanL) caused by Leishmania infantum is a widespread endemic disease in SW Europe. This study was designed to determine how veterinarians clinically manage CanL in this region by analysing information collected in a questionnaire completed by local veterinarians working in clinics in France, Portugal, Greece, Spain, Italy and Slovenia.MethodsOver the period 2004–2011, a questionnaire on CanL was sent to 12,546 small animal clinics located in the six countries surveyed. The questionnaire with 10 items comprising open and closed questions sought to obtain comparable data regarding the main clinical manifestations of CanL, the diagnostic methods used, the treatment regimens selected, recommended preventive measures and awareness of the important public health implications of CanL.ResultsThe data collected reflect similarities in the clinical manifestations reported although there was some variation in the concurrent diseases described, and wide variation in the clinical management of CanL among the countries examined in terms of dosing regimens, therapeutic agents and the criteria used to diagnose CanL. Most veterinarians properly informed dog owners about the preventive measures available and about the zoonotic implications of CanL.ConclusionsThis survey describes the current situation in SW endemic countries in Europe regarding the clinical management of CanL. The data collected reveal a need to unify criteria from evidence-based medicine to determine and similarly apply the best diagnostic and treatment methods available for this disease in the different countries.

First Report of 13 Species of Culicoides (Diptera: Ceratopogonidae) in Mainland Portugal and Azores by Morphological and Molecular Characterization

The genus Culicoides (Diptera: Ceratopogonidae) contains important vectors of animal and human diseases, including bluetongue, African horse sickness and filariosis. A major outbreak of bluetongue occurred in mainland Portugal in 2004, forty eight years after the last recorded case. A national Entomological Surveillance Plan was initiated in mainland Portugal, Azores and the Madeira archipelagos in 2005 in order to better understand the disease and facilitate policy decisions. During the survey, the most prevalent Culicoides species in mainland Portugal was C. imicola (75.3%) and species belonging to the Obsoletus group (6.5%). The latter were the most prevalent in Azores archipelago, accounting for 96.7% of the total species identified. The Obsoletus group was further characterized by multiplex Polymerase Chain Reaction to species level showing that only two species of this group were present: C. obsoletus sensu strictu (69.6%) and C. scoticus (30.4%). Nine species of Culicoides were detected for the first time in mainland Portugal: C. alazanicus, C. bahrainensis, C. deltus, C. lupicaris, C. picturatus, C. santonicus, C. semimaculatus, C. simulator and C. subfagineus. In the Azores, C. newsteadi and C. circumscriptus were identified for the first time from some islands, and bluetongue vectors belonging to the Obsoletus group (C. obsoletus and C. scoticus) were found to be widespread.

Range expansion of the Bluetongue vector, Culicoides imicola , in continental France likely due to rare wind-transport events

The role of the northward expansion of Culicoides imicola Kieffer in recent and unprecedented outbreaks of Culicoides-borne arboviruses in southern Europe has been a significant point of contention. We combined entomological surveys, movement simulations of air-borne particles, and population genetics to reconstruct the chain of events that led to a newly colonized French area nestled at the northern foot of the Pyrenees. Simulating the movement of air-borne particles evidenced frequent wind-transport events allowing, within at most 36 hours, the immigration of midges from north-eastern Spain and Balearic Islands, and, as rare events, their immigration from Corsica. Completing the puzzle, population genetic analyses discriminated Corsica as the origin of the new population and identified two successive colonization events within west-Mediterranean basin. Our findings are of considerable importance when trying to understand the invasion of new territories by expanding species.