a retrospective study of eighteen years

Leptospirosis, an under-recognized public health problem, needs to be confirmed through specific laboratory diagnosis. We describe herein a series of 4618 symptomatic patients for whom a microagglutination test (MAT) serology was available, representing a unique picture of human leptospirosis in central mainland Portugal and the Azores islands of São Miguel and Terceira, over eighteen- and twelve-year periods, respectively. The distribution of the 1024 (22%) cases identified was an average 57 cases per year, with higher frequency in males (67%). These represent the majority of leptospirosis notifications in Portugal, with a higher annual incidence rate in the islands, compared to the central mainland (11.1 and 1.7/100,000 population, respectively). Middle-aged adults (25-54 years) were most frequently infected (45%). Cases occurred mainly in December and January. Serovars from nine presumptive serogroups caused infection, with a predominance of Icterohaemorrhagiae, Pomona, and Ballum, accounting for 66% of cases. Seropositivity was associated with both anicteric and icteric leptospirosis. Several risk factors and a higher transmission risk in certain areas were emphasized. Leptospirosis was confirmed as a steadily increasing public health problem; good surveillance, communication, and laboratory support are thus necessary to reduce the impact of leptospirosis in areas at risk.publishersversionpublishe

Human leptospirosis in Portugal: a retrospective study of eighteen years

SummaryIntroductionLeptospirosis, an under-recognized public health problem, needs to be confirmed through specific laboratory diagnosis.DesignWe describe herein a series of 4618 symptomatic patients for whom a microagglutination test (MAT) serology was available, representing a unique picture of human leptospirosis in central mainland Portugal and the Azores islands of São Miguel and Terceira, over eighteen- and twelve-year periods, respectively.ResultsThe distribution of the 1024 (22%) cases identified was an average 57 cases per year, with higher frequency in males (67%). These represent the majority of leptospirosis notifications in Portugal, with a higher annual incidence rate in the islands, compared to the central mainland (11.1 and 1.7/100000 population, respectively). Middle-aged adults (25–54 years) were most frequently infected (45%). Cases occurred mainly in December and January. Serovars from nine presumptive serogroups caused infection, with a predominance of Icterohaemorrhagiae, Pomona, and Ballum, accounting for 66% of cases.ConclusionsSeropositivity was associated with both anicteric and icteric leptospirosis. Several risk factors and a higher transmission risk in certain areas were emphasized. Leptospirosis was confirmed as a steadily increasing public health problem; good surveillance, communication, and laboratory support are thus necessary to reduce the impact of leptospirosis in areas at risk

que apoio ao diagnóstico de Borreliose de Lyme?

No diagnóstico de borreliose de Lyme, infecção sub-diagnosticada em Portugal, o apoio laboratorial é crucial para o tratamento atempado dos doentes, sob risco destes evoluírem para fases crónicas, resistentes à terapêutica. A difícil confirmação do agente etiológico através da cultura, aliada ao conhecimento actual sobre as elevadas taxas de falsos negativos nos testes serológicos de rastreio, bem como de casos seronegativos de doença, torna da maior importância a implementação da nova tecnologia de amplificação (PCR) de DNA borreliano no diagnóstico de rotina desta patologia. Tratando-se de uma técnica mais rápida e sensível tem-se revelado de maior sucesso do que a cultura. O presente trabalho compara a sensibilidade destas duas técnicas de detecção directa dos agentes do complexo Borrelia burgdorferi sensu lato, em 96 doentes com suspeita clínica desta afecção e em 569 vectores (ixodídeos) capturados em território nacional. Obteve-se o isolamento de borrélias em 1,3% das biópsias cutâneas e em 1,2% dos ixodídeos (carraças) não se tendo registado isolados noutros materiais biológicos humanos. A aplicação da técnica de nested PCR confirmou a presença de DNA de B. burgdorferi sensu lato em 24,8% das amostras analisadas. A identificação dos agentes patogénicos, por técnicas de hibridação (Reverse Line Blot), sequenciação e/ou RFLP, revelou a presença de quatro espécies genómicas: B. garinii, B. lusitaniae, B. valaisiana e B. afzelii. Em conclusão, a amplificação de DNA borreliano permite uma resposta mais rápida e sensível do que a cultura, contribuindo para um diagnóstico laboratorial mais eficaz da borreliose de Lyme humana. Lyme borreliosis (LB) is an under-diagnosed zoonosis in Portugal, specially due to the absence of specific clinical signs. The role of the laboratory diagnosis, together with an epidemiological information, is extremely important for the correct treatment of patients with this pathology. The sensitivity of two laboratory techniques (culture and DNA amplification by nested PCR) for direct detection of Borrelia burgdorferi sensu lato complex agents was evaluated in samples from 96 clinically suspected patients, as also in 569 vector ticks collected throughout Portugal. Borrelia genospecies were identified in 1.3% of the skin biopsies and in 1.2% of the vectors, after growth in selective culture medium (BSK). No growth was obtained from other type of samples. B.burgdorferi sensu lato DNA was present in 24.8% of analyzed samples, as per intergenic rRNA 5S-23S (rrf–rrl) spacer amplification by nested PCR. Four genomic species (B.garinii, B.lusitaniae, B.valaisiana and B.afzelii) were identified by DNA hybridization (Reverse-Line-Blot), sequencing and/or RFLP. In conclusion and taking into account the results found in this study, it seems that Borrelia DNA amplification technique is quicker and present higher sensitivity than culture, providing a more effective laboratory diagnosis of LB in human populations and in vectors.publishe

Bone length of lberian freshwater fish, as a predictor of length and biomass of prey consumed by piscivores

We measured various fish bones from 13 Iberian freshwater fish species and one hybrid species. Original total body lengths were then back-calculated using bone measurements. Bones usually found in prey remains left by piscivorous predators, were usually from the head skeleton and from the vertebral column. The 73 regressions obtained between bone length and fish length were linear for al1 species examined. Coefficients of determination ranged between 75.6 % and 99.5 %. To estimate fish biomass, length-weight relations were used for each species, too. Bone length vs. fish length relationships found in the literature for seven additional species inhabiting the Iberian Peninsula were also included, totalling 29 regression equations. The amount of dietary information available from fish predator remains can be greatly increased by using these relationships. In this paper, information is provided covering in excess of 37 YO of lberian freshwater fish fauna, including the most abundant and widespread species.-----------------------------------------------------------------------Se han realizado medidas a 13 tipos de huesos de peces para retrocalcular su longitud total. Estos huesos, que se encuentran normalmente en los restos dejados por depredadores ictiófagos, pertenecen al esqueleto de la cabeza y a la columna vertebral de 13 especies de peces (más un híbrido) de aguas continentales de la Península Ibérica. Las 73 ecuaciones obtenidas entre la longitud del hueso y la del pez fueron lineales para todas las especies estudiadas; los coeficientes de determinación oscilaron entre el 75.6 %y el 99.5 %. Para la estimación de la biomasa de los peces se han calculado las regresiones longitud- peso para cada una de las especies. Además, se incluyen, extraídas de la bibliogra$a, las relaciones entre la longitud del hueso y la longitud del pez para otras siete especies de peces que habitan en la Península Ibérica. Con este método la cantidad de información extraíble del análisis de la dieta de depredadores ictiófagos se puede incrementar considerablemente. Las ecuaciones que se aportan abarcan a más del 37 96 de la ictiofauna Ibérica de aguas continentales, con inclusión de las especies más abundantes y de más amplia distribución

First isolation of human Leptospira strains, Azores, Portugal.

OBJECTIVES The aim of this study was the first identification of Leptospira isolates from Azorean inpatients. METHODS Whole blood samples from 68 inpatients attending the São Miguel Hospital between 2006 and 2008, with a clinical and epidemiological suspicion of leptospirosis, were inoculated in a transport medium broth at the patient's bedside and further processed using a serial dilution technique prior to culture. At admission, 62 (91%) patients were also analyzed for the presence of leptospiral DNA by a nested PCR and 40 (59%) for specific agglutinins by microscopic agglutination test (MAT). The isolates obtained were first assigned at the serogroup level by both MAT reactivity with hyperimmune rabbit antisera and a PCR-based assay with the single primer iRep1. The species identification was performed by DNA sequencing. The use of monoclonal antibodies allowed intraspecific discrimination at the serovar level. RESULTS Of the 10 (14.7%) human Leptospira isolates, seven were identified as Leptospira interrogans serovar Copenhageni and three as Leptospira borgpetersenii serovar Arborea, which is in agreement with previous data from the Azorean rodent population. CONCLUSIONS This study represents a great step towards the definitive identification of the pathogenic leptospires in Azorean patients and confirms the bacteriological human-rodent connection for the first time.publishersversionpublishe

Fine-Scale Phylogeographic Structure of Borrelia lusitaniae Revealed by Multilocus Sequence Typing

Borrelia lusitaniae is an Old World species of the Lyme borreliosis (LB) group of tick-borne spirochetes and prevails mainly in countries around the Mediterranean Basin. Lizards of the family Lacertidae have been identified as reservoir hosts of B. lusitaniae. These reptiles are highly structured geographically, indicating limited migration. In order to examine whether host geographic structure shapes the evolution and epidemiology of B. lusitaniae, we analyzed the phylogeographic population structure of this tick-borne bacterium using a recently developed multilocus sequence typing (MLST) scheme based on chromosomal housekeeping genes. A total of 2,099 questing nymphal and adult Ixodes ricinus ticks were collected in two climatically different regions of Portugal, being ∼130 km apart. All ticks were screened for spirochetes by direct PCR. Attempts to isolate strains yielded 16 cultures of B. lusitaniae in total. Uncontaminated cultures as well as infected ticks were included in this study. The results using MLST show that the regional B. lusitaniae populations constitute genetically distinct populations. In contrast, no clear phylogeographic signals were detected in sequences of the commonly used molecular markers ospA and ospC. The pronounced population structure of B. lusitaniae over a short geographic distance as captured by MLST of the housekeeping genes suggests that the migration rates of B. lusitaniae are rather low, most likely because the distribution of mediterranean lizard populations is highly parapatric. The study underlines the importance of vertebrate hosts in the geographic spread of tick-borne microparasites

Seed storage conditions change the germination pattern of clonal growth plants in Mediterranean salt marshes.

8 páginas, 4 tablas, 8 figuras.The effect of salinity level and extended exposure to different salinity and flooding conditions on germination patterns of three saltmarsh clonal growth plants (Juncus subulatus, Scirpus litoralis, and S. maritimus) was studied. Seed exposure to extended flooding and saline conditions significantly affected the outcome of the germination process in a different, though predictable, way for each species, after favorable conditions for germination were restored. Tolerance of the germination process was related to the average salinity level measured during the growth/germination season at sites where established individuals of each species dominated the species cover. No relationship was found between salinity tolerance of the germination process and seed response to extended exposure to flooding and salinity conditions. The salinity response was significantly related to the conditions prevailing in the habitats of the respective species during the unfavorable (nongrowth/nongermination) season. Our results indicate that changes in salinity and hydrology while seeds are dormant affect the outcome of the seed-bank response, even when conditions at germination are identical. Because these environmental-history-dependent responses differentially affect seed germination, seedling density, and probably sexual recruitment in the studied and related species, these influences should be considered for wetland restoration and managementFinancial support from the Spanish Ministry of the Environment (MMA, project 05/99) and the Junta de Andalucía (research group 4086)enabled us to carry out the present work.Peer reviewe

Seed storage conditions change the germination pattern of clonal growth plants in Mediterranean salt marshes.

8 páginas, 4 tablas, 8 figuras.The effect of salinity level and extended exposure to different salinity and flooding conditions on germination patterns of three saltmarsh clonal growth plants (Juncus subulatus, Scirpus litoralis, and S. maritimus) was studied. Seed exposure to extended flooding and saline conditions significantly affected the outcome of the germination process in a different, though predictable, way for each species, after favorable conditions for germination were restored. Tolerance of the germination process was related to the average salinity level measured during the growth/germination season at sites where established individuals of each species dominated the species cover. No relationship was found between salinity tolerance of the germination process and seed response to extended exposure to flooding and salinity conditions. The salinity response was significantly related to the conditions prevailing in the habitats of the respective species during the unfavorable (nongrowth/nongermination) season. Our results indicate that changes in salinity and hydrology while seeds are dormant affect the outcome of the seed-bank response, even when conditions at germination are identical. Because these environmental-history-dependent responses differentially affect seed germination, seedling density, and probably sexual recruitment in the studied and related species, these influences should be considered for wetland restoration and managementFinancial support from the Spanish Ministry of the Environment (MMA, project 05/99) and the Junta de Andalucía (research group 4086)enabled us to carry out the present work.Peer reviewe

Doñana. Acta vertebrata. vol 24(1/2)

Alimentación de la lagartija colilarga Psammodromus algirus (L) (Sauria, Lacertidae), en el litoral de Huelva (SO EspañaLa alimentación de Myotis myotis Borkh, 1791 (Chiroptera, Vespertilionidae) en la cuenca del rio Guadix (sureste de España)Distribución y selección de hábitat de la garduña (Martes foina, Erxleben, 1777) en Vizcaya y Sierra Salvada (Burgos).Nuevo modelo de trampa para reducir el impacto de la pesca de cangrejos sobre los vertebrados en las marismas del GuadalquiviThe functions of song and the spatial pattern of song production in the rufous bush chat (Cercotrichas galactotes)Migración e invernada de las lavanderas cascadena Motacilla cinerea y blanca M. alba en la Penínula Ibérica e Islas BalearesAbundancia y reproducción de Glis glis (Linnaeus, 1766) (Rodentia, Gliridae) en el Pirineo occidental.Estatus de residencia, categorización trófica y abundancia de aves en el zoológico de La Plata, ArgentinaAvifauna reproductora y estructura del hábitat en la campiña y sierras Subbéticas de JaénDispersión de semillas de retama (Retama sphaerocarpa (L.) Boiss por el conejo (Oryctolagus cuniculus L.) en el centro de EspañaGuía para la identificación de restos óseos pertenecientes a algunos peces comunes en las aguas continentales de la Península Ibérica para el estudio de la dieta de depredadores ictiófagosDistribución y abundancia del corzo (Capreolus capreolus L. 1758) en la provincia de JaénAlimentación de las larvas de anuros en ambientes temporales del sistema del rio Paraná, Argentina.Mauremys leprosa como presa de Lutra lutraNota sobre la alimentación del lince ibérico en el Parque Natural de la Sierra de Andújar (Sierra Morena oriental)Presencia de Echinococcus granulosus (Cestoda) en un lobo ibérico (Canis lupusLa orientación de los nidos de paseriformes estepariosComparación de la dieta obtenida a partir de muestras estomacales y fecales del Tuco-tuco, Ctenomys mendocinus, en dos poblaciones de la precordillera de los Andes, ArgentinaEstructura genética y distribución de la variabilidad enzimática en poblaciones naturales de estornino negro (Sturnus unicolor)Estimación de la disponibilidad trófica para el quebrantahuesos (Gypaetus barbatus) en Cataluña (NE España) e implicaciones sobre su conservaciónPeer reviewe

Contribuicao para o estudo do genero Leptospira em Portugal

The spirochetes of the genus Leptospira are the aetiological agents of a worlwide zoonosis which causes important problems in Public Health and in livestock. Owing to the very incomplete knowledge on the biology and epidemiology of leptospires in Portugal, the goal of this thesis was to contribute to the evaluation of some of those aspects. Altogether, the identification of the pathogenic serogroups of the human and animal infections, the sequence of the IgM and IgC immunoglobulins production during the disease and some aprticular genetic aspects concerning the plasmid detection, deserved a special attention. The human immunological surveys were developed by the microscopic agglutination test in an hospitalized population (1677 inpatients mainly from hospitals in both districts of Lisbon and Coimbra), in a high risk population (346 abattoir workers) and in an healthy population (304 rural residents). Among other observations, it was confirmed the occurence of leptospirosis in the examined inpatients (21.3%) and the existence of a significant variation of the pathogenic serogroup, according to their urban and rural origin. A reduced leptospiral prevalence in the high risk groups (2.4% and 5.2%) and a percentage of 3.0% seropositives within healthy subjects were noticed. The search of anti-Leptospira agglutinins in animals confirmed the occurence of leptospirosis in all geographic regions of the country and in all livetock species, although the most significative prevalence was found within swines (32,3%) and bovines (23.3%). The isolation of hardjo serovar in bovines is emphasized as it is an important causal agent of severe reproduction hazards and milk production losses. Besides the relevant seropositivity observed in dogs (32.1%), it was also described the first detection of specific anti-body in wolves and foxes. The prospective study of the humoral immunity in 237 sera from 77 leptospirosis cases showed the epidemiological value of the ELISA technique in serological surveys, both as a confirmatory test of recent Leptospira infections when there are single titres (ELISA-IgM)equal or greater than 1:1000, and as a confirmatory test of the infecting serogroup in residual infections (ELISA-IgG). Finally, the already suggested absence of short sized plasmids in the genus Leptospira, as the result of a preliminary survey in only or three serovars, was now confirmed by the plasmidic research on 73 pathogenic and saprophytic strains. In conclusion, in this work the importance of leptospirosis in Portugal is emphasized, in order to recommend a careful clinical attention and a more sensitive and specific laboratorial diagnosis, considering the great number of inespecific clinical features of the disease, both in urban and rural centersAvailable from Fundacao para a Ciencia e a Tecnologia, Servico de Informacao e Documentacao, Av. D. Carlos I, 126, 1200 Lisboa / FCT - Fundação para o Ciência e a TecnologiaSIGLEPTPortuga