Experimental study of the susceptibility of a European Aedes albopictus strain to dengue virus under a simulated Mediterranean temperature regime.

Dengue virus (DENV) has re-emerged in Europe driven by the geographic expansion of the mosquito species Aedes albopictus and Aedes aegypti (Diptera: Culicidae) and the introduction of the virus by viraemic travellers. In the present study, the vector competence (VC) of Ae. albopictus collected in Catalonia (northeast Spain) was evaluated for two different DENV strains, DENV-1 and DENV-2, the serotypes responsible for all outbreaks of dengue that have occurred in Europe. Mosquitoes were reared under environmental conditions mimicking the mean temperature and humidity recorded in July on the Mediterranean coast of Catalonia. Mosquitoes were fed on an artificial infectious bloodmeal and, at 14 days post-exposure, infection, disseminated infection and transmission rates (IR, DIR, TR) and transmission efficiency (TE) were determined by testing the virus in the body, legs and saliva. The tested Ae. albopictus strain was found to be susceptible to both DENV-1 and DENV-2 and to be able to transmit DENV-1. This is the first time that the VC of Ae. albopictus for DENV has been tested in Europe in this specific context (i.e. mimicking the Mediterranean temperature and humidity recorded in Catalonia in July). This study confirms the potential of Ae. albopictus to start autochthonous DENV transmission cycles in the Mediterranean basin.

Transmission and immunopathology of the avian influenza virus A/Anhui/1/2013 (H7N9) human isolate in three commonly commercialized avian species.

H7N9 virus infection is a global concern, given that it can cause severe infection and mortality in humans. However, the understanding of H7N9 epidemiology, animal reservoir species and zoonotic risk remains limited. This work evaluates the pathogenicity, transmissibility and local innate immune response of three avian species harbouring different respiratory distribution of α2,6 and α2,3 SA receptors. Muscovy ducks, European quails and SPF chickens were intranasally inoculated with 105 embryo infectious dose (EID)50 of the human H7N9 (A/Anhui/1/2013) influenza isolate. None of the avian species showed clinical signs or macroscopic lesions, and only mild microscopic lesions were observed in the upper respiratory tract of quail and chickens. Quail presented more severe histopathologic lesions and avian influenza virus (AIV) positivity by immunohistochemistry (IHC), which correlated with higher IL-6 responses. In contrast, Muscovy ducks were resistant to disease and presented higher IFNα and TLR7 response. In all species, viral shedding was higher in the respiratory than in the digestive tract. Higher viral shedding was observed in quail, followed by chicken and ducks, which presented similar viral titres. Efficient transmission was observed in all contact quail and half of the Muscovy ducks, while no transmission was observed between chicken. All avian species showed viral shedding in drinking water throughout infection.

Rift Valley fever virus and European mosquitoes: vector competence of Culex pipiens and Stegomyia albopicta (= Aedes albopictus).

Rift Valley fever (RVF) is a mosquito-borne disease caused by the Rift Valley fever virus (RVFV). Rift Valley fever affects a large number of species, including human, and has severe impact on public health and the economy, especially in African countries. The present study examined the vector competence of three different European mosquito species, Culex pipiens (Linnaeus, 1758) form molestus (Diptera: Culicidae), Culex pipiens hybrid form and Stegomyia albopicta (= Aedes albopictus) (Skuse, 1894) (Diptera: Culicidae). Mosquitoes were artificially fed with blood containing RVFV. Infection, disseminated infection and transmission efficiency were evaluated. This is the first study to assess the transmission efficiency of European mosquito species using a virulent RVFV strain. The virus disseminated in Cx. pipiens hybrid form and in S. albopicta. Moreover, infectious viral particles were isolated from saliva of both species, showing their RVFV transmission capacity. The presence of competent Cx. pipiens and S. albopicta in Spain indicates that an autochthonous outbreak of RVF may occur if the virus is introduced. These findings provide information that will help health authorities to set up efficient entomological surveillance and RVFV vector control programmes.

Epidemiological and pathological investigation of fowl aviadenovirus serotypes 8b and 11 isolated from chickens with inclusion body hepatitis in Spain (2011-2013).

Inclusion body hepatitis caused by different fowl aviadenovirus (FAdV) serotypes has been described in several countries in recent years. In Spain, from the spring of 2011 to 2013, an increased number of outbreaks in broiler and broiler breeder flocks from different regions occurred. The objectives of the present work were to carry out the molecular characterization of FAdV strains from Spanish inclusion body hepatitis cases and to study the pathogenicity and viral dynamics of these strains in specific pathogen-free (SPF) chickens. A total of 52 inclusion body hepatitis clinical cases, including 45 from broiler farms and seven from broiler breeder farms, were analysed by conventional polymerase chain reaction and sequencing targeting the FAdV hexon gene. From these, 37 strains were classified as FAdV type 8b, while the remaining 15 were classified as FAdV types 11 (n = 10), 2 (n = 4) and 8a (n = 1). In addition, two different FAdVs belonging to the genotypes 8b and 11 were used for experimental infection. Specific pathogen-free five-day-old birds were inoculated intramuscularly with a high (106.5 tissue culture infective dose (TCID)50/ml) or low (104 TCID50/ml) dose of the above-mentioned FAdVs. No mortality was observed in any of the experimental groups, and only one bird showed evident clinical signs. However, macroscopic and microscopic hepatic lesions, as well as viral DNA, were detected in birds from all infection groups. Inclusion bodies and viral DNA were also detected in the pancreas and in the small and the large intestine in some birds. Long-lasting shedding and transmission to contact birds were confirmed in all infected groups.

Influenza A Virus Surveillance in the Invasive American Mink (Neovison vison) from Freshwater Ecosystems, Northern Spain.

Influenza A viruses (IAVs) are negative-sense, single-stranded and segmented RNA viruses of the Orthomyxoviridae family that may cause acute respiratory disease in a wide range of birds and mammals. Susceptibility of several species within the family Mustelidae to IAVs has been reported as a result of natural or experimental infections. The objectives of this study were to assess whether free-ranging American mink populations from Northern Spain were infected with IAV and try to define the role of this species in the epidemiology of IAV. Sera from 689 American mink from Northern Spain captured between 2011 and 2014 were tested for the presence of antibodies against IAVs using a commercial competition cELISA. Positive sera were further analysed with haemagglutination inhibition (HI) assay. Fifteen of the 689 (2.2%, 1.3-3.6 CI95% ) of the American minks analysed were ELISA positive. No significant differences were observed between years of capture, provinces, river basins, sexes or ages of the animals. All seropositive sera resulted negative to the panel strains used in the HI assay, showing that the most relevant strains circulating in swine, the most relevant avian subtypes (H5 and H7) and the H10N4 subtype isolated in minks have not been circulating in this free-ranging exotic carnivore from Spain. In the light of these results, the free-range American mink from Northern Spain do not seem to have an important role in the epidemiology of IAVs.

Survey of laboratory-acquired infections around the world in biosafety level 3 and 4 laboratories.

Laboratory-acquired infections due to a variety of bacteria, viruses, parasites, and fungi have been described over the last century, and laboratory workers are at risk of exposure to these infectious agents. However, reporting laboratory-associated infections has been largely voluntary, and there is no way to determine the real number of people involved or to know the precise risks for workers. In this study, an international survey based on volunteering was conducted in biosafety level 3 and 4 laboratories to determine the number of laboratory-acquired infections and the possible underlying causes of these contaminations. The analysis of the survey reveals that laboratory-acquired infections have been infrequent and even rare in recent years, and human errors represent a very high percentage of the cases. Today, most risks from biological hazards can be reduced through the use of appropriate procedures and techniques, containment devices and facilities, and the training of personnel.

Culex pipiens and Stegomyia albopicta (= Aedes albopictus) populations as vectors for lineage 1 and 2 West Nile virus in Europe.

The emerging disease West Nile fever is caused by West Nile virus (WNV), one of the most widespread arboviruses. This study represents the first test of the vectorial competence of European Culex pipiens Linnaeus 1758 and Stegomyia albopicta (= Aedes albopictus) (both: Diptera: Culicidae) populations for lineage 1 and 2 WNV isolated in Europe. Culex pipiens and S. albopicta populations were susceptible to WNV infection, had disseminated infection, and were capable of transmitting both WNV lineages. This is the first WNV competence assay to maintain mosquito specimens under environmental conditions mimicking the field (day/night) conditions associated with the period of maximum expected WNV activity. The importance of environmental conditions is discussed and the issue of how previous experiments conducted in fixed high temperatures may have overestimated WNV vector competence results with respect to natural environmental conditions is analysed. The information presented should be useful to policymakers and public health authorities for establishing effective WNV surveillance and vector control programmes. This would improve preparedness to prevent future outbreaks.

Ecological surveillance for West Nile in Catalonia (Spain), learning from a five-year period of follow-up.

To enhance early detection of West Nile virus (WNV) transmission, an integrated ecological surveillance system was implemented in Catalonia (north-eastern Spain) from 2007 to 2011. This system incorporated passive and active equine surveillance, periodical testing of chicken sentinels in wetland areas, serosurveillance wild birds and testing of adult mosquitoes. Samples from 298 equines, 100 sentinel chickens, 1086 wild birds and 39 599 mosquitoes were analysed. During these 5 years, no acute WNV infection was detected in humans or domestic animal populations in Catalonia. WNV was not detected in mosquitoes either. Nevertheless, several seroconversions in resident and migrant wild birds indicate that local WNV or other closely related flaviviruses transmission was occurring among bird populations. These data indicate that bird and mosquito surveillance can detect otherwise silent transmission of flaviviruses and give some insights regarding possible avian hosts and vectors in a European setting.

Lymphoplasmacytic endotheliitis and anterior uveitis in sheep infected experimentally with rift valley fever virus.

Lymphoplasmacytic endotheliitis and anterior uveitis was diagnosed in four lambs infected experimentally with field isolates of Rift Valley fever virus (RVFV). Formalin-fixed and paraffin wax-embedded tissue from these animals was investigated by histopathology and quantitative real time reverse transcriptase polymerase chain reaction. To our knowledge, this is the first pathological description of this ocular manifestation of RVFV infection in ruminants, although these lesions have been described in man.

Detection and monitoring of mosquito flaviviruses in Spain between 2001 and 2005.

Between the years 2001 and 2005, a total of 72,895 female mosquitoes were trapped during their season of abundance, and analyzed. They were sorted into 4,723 pools belonging to 20 Culicidae species from the Anopheles, Aedes, Ochlerotatus, Culex, Culiseta, Coquillettidia, and Uranotaenia genera. The aim was to detect arboviral RNA directly from mosquito homogenates for the genera Alphavirus, Flavivirus, and Phlebovirus. The study formed part of general arbovirus transmission research in four of the most important wetlands in Spain; in the provinces of Girona, Barcelona, Tarragona, and Huelva. The mosquitoes were collected using human bait, CO(2) traps, or light traps, and they were pooled according to date of collection, location, and species. No arboviral RNA from known pathogenic arboviruses was found. However, 111 pools tested positive for unknown mosquito Flavivirus, the only genus detected. The Flavivirus sequences identified were different from all known Flavivirus mosquito viruses, but very close to Kamiti River virus or cell fusing agent virus. The maximum likelihood estimation infection rate (MLE) was calculated for all regions and species. Aedes albopictus had the highest MLE at 47.14, followed by Ae. vexans with 43.67 (over the entire area). These species were followed by Culiseta annulata, with 36.00. The most common species, Ochlerotatus caspius and Culex pipiens, had low MLE values-0.94 and 0.38, respectively-over the area as a whole.

[Streptococcus suis meningitis in an immunocompetent patient]. / Meningitis por Streptococcus suis en un paciente inmunocompetente.

A case of Streptococcus suis meningitis is described in an immunocompetent patient presenting asthenia, general weakness, fever, vomiting, sensory deterioration and temporospatial disorder. The cerebrospinal fluid and two blood cultures (2/2 bottles) were positive. The isolate was preliminary identified by conventional biochemical tests, and the identification was completed at the Special Bacteriology Service of INEI-ANLIS "Dr. Carlos G. Malbrán". Ampicillin and ceftriaxone treatment was initiated. The isolate was susceptible to ampicillin, cefotaxime and vancomycin. The patient experienced a good outcome but suffered hearing loss. However, after four months he returned with walking ataxia, deafness in his left ear, and hearing loss in the right ear. The patient's retrospective exposure to pigs had been verified. It is important to evaluate predisposing and epidemiologic factors in order to alert about the possible presence of this etiological agent in cases of meningitis or bacteremia.

Meningitis por Streptococcus suis en un paciente inmunocompetente / Streptococcus suis meningitis in an immunocompetent patient

Se describe un caso de meningitis por Streptococcus suis en un paciente inmunocompetente. Presentaba astenia, debilidad generalizada, fiebre (39 °C), vómitos, deterioro del sensorio y desorientación témporo-espacial. Los cultivos de sangre (2/2) y de líquido cefalorraquídeo fueron positivos. La identificación preliminar se realizó utilizando las pruebas bioquímicas convencionales y fue completada en el Servicio Bacteriología Especial del INEI-ANLIS "Dr. Carlos G. Malbrán". Se comenzó el tratamiento con ampicilina y ceftriaxona. El microorganismo aislado demostró sensibilidad a ampicilina, cefotaxima y vancomicina. El paciente evolucionó favorablemente, pero se comprobó leve hipoacusia. Reingresó a los 4 meses con marcha atáxica, anacusia en oído izquierdo e hipoacusia en oído derecho. Continúa con seguimiento neurológico y audiométrico. Retrospectivamente se constató el contacto del paciente con cerdos. Se destaca la importancia de la anamnesis para alertar la sospecha de este agente etiológico en meningitis y bacteriemias.

A case of Streptococcus suis meningitis is described in an immunocompetent patient presenting asthenia, general weakness, fever, vomiting, sensory deterioration and temporospatial disorder. The cerebrospinal fluid and two blood cultures (2/2 bottles) were positive. The isolate was preliminary identified by conventional biochemical tests, and the identification was completed at the Special Bacteriology Service of INEI-ANLIS "Dr. Carlos G. Malbrán". Ampicillin and ceftriaxone treatment was initiated. The isolate was susceptible to ampicillin, cefotaxime and vancomycin. The patient experienced a good outcome but suffered hearing loss. However, after four months he returned with walking ataxia, deafness in his left ear, and hearing loss in the right ear. The patient’s retrospective exposure to pigs had been verified. It is important to evaluate predisposing and epidemiologic factors in order to alert about the possible presence of this etiological agent in cases of meningitis or bacteremia.

[Isolated pathogen microorganisms in respiratory samples from children with cystic fibrosis]. / Microorganismos patógenos aislados en muestras respiratorias de niños con fibrosis quística.

Cystic Fibrosis (CF) is characterized by a dysfunction of the exocrine secretion glands. The first symptoms often appear in the respiratory system which constitutes one of the most important morbimortality causes in these patients. Chronic respiratory tract colonization is caused mainly by bacteria such as Staphylococcus aureus, Haemophilus spp. and Pseudomonas aeruginosa. Respiratory samples from patients with CF (age group: 4 months to 11 years) were analyzed from November 2001 to August 2004. The most frequently isolated microorganisms were S. aureus (38.7%), P. aeruginosa (37.4%) and Haemophilus spp (15.3%). A high resistance to erithromycine (35.0%) and clindamicine (29.4%) was observed in S. aureus strains and 25.9% of them were methicillin-resistant. P. aeruginosa strains were mainly gentamicin-resistant (31.0%). The rate of ampicillin-resistant Haemophilus spp. was 23.0% and it was due to the presence of beta-lactamases, but a high trimethoprim-sulfamethoxazole resistance was observed in this microorganism (59.0%).

Microorganismos patógenos aislados en muestras respiratorias de niños con fibrosis quística / Isolated pathogen microorganisms in respiratory samples from children with cystic fibrosis

La fibrosis quística (FQ) se caracteriza por disfunciones en las glándulas de secreción exocrina del organismo. Las primeras manifestaciones suelen observarse en el sistema respiratorio, constituyendo una de las causas más importantes de morbimortalidad en los pacientes afectados. Los microorganismos patógenos que colonizan frecuentemente el tracto respiratorio de estos pacientes son Staphylococcus aureus, Haemophilus spp., y Pseudomonas aeruginosa. Entre noviembre de 2001 y agosto de 2004 se estudiaron 222 muestras respiratorias de pacientes con FQ de entre 4 meses y 11 años de edad. Se aislaron S. aureus (38,7%), P. aeruginosa (37,4%) y Haemophilus spp., (15,3%). En S. aureus la meticilina-resistencia fue del 25,9% y se asoció con altas resistencias a eritromicina (35,0%) y clindamicina (29,4%). El mayor porcentaje de resistencia observado en las cepas de P. aeruginosa fue frente a gentamicina (31,0%). Los aislamientos de Haemophilus spp. fueron resistentes a ampicilina (23,0%) debido a la presencia de beta-lactamasas, y a trimetoprima/sulfametoxazol (59,0%).

Cystic Fibrosis (CF) is characterized by a dysfunction of the exocrine secretion glands. The first symptoms often appear in the respiratory system which constitutes one of the most important morbimortality causes in these patients. Chronic respiratory tract colonization is caused mainly by bacteria such as Staphylococcus aureus, Haemophilus spp. and Pseudomonas aeruginosa. Respiratory samples from patients with CF (age group: 4 months to 11 years) were analyzed from November 2001 to August 2004. The most frequently isolated microorganisms were S. aureus (38.7%), P. aeruginosa (37.4%) and Haemophilus spp (15.3%). A high resistance to erithromycine (35.0%) and clindamicine (29.4%) was observed in S. aureus strains and 25.9% of them were methicillin-resistant. P. aeruginosa strains were mainly gentamicin-resistant (31.0%). The rate of ampicillin-resistant Haemophilus spp. was 23.0% and it was due to the presence of beta-lactamases, but a high trimethoprim-sulfamethoxazole resistance was observed in this microorganism (59.0%).

Microorganismos patógenos aislados en muestras respiratorias de niños con fibrosis quística. / [Isolated pathogen microorganisms in respiratory samples from children with cystic fibrosis]

Cystic Fibrosis (CF) is characterized by a dysfunction of the exocrine secretion glands. The first symptoms often appear in the respiratory system which constitutes one of the most important morbimortality causes in these patients. Chronic respiratory tract colonization is caused mainly by bacteria such as Staphylococcus aureus, Haemophilus spp. and Pseudomonas aeruginosa. Respiratory samples from patients with CF (age group: 4 months to 11 years) were analyzed from November 2001 to August 2004. The most frequently isolated microorganisms were S. aureus (38.7

), P. aeruginosa (37.4

) and Haemophilus spp (15.3

). A high resistance to erithromycine (35.0

) and clindamicine (29.4

) was observed in S. aureus strains and 25.9

of them were methicillin-resistant. P. aeruginosa strains were mainly gentamicin-resistant (31.0

). The rate of ampicillin-resistant Haemophilus spp. was 23.0

and it was due to the presence of beta-lactamases, but a high trimethoprim-sulfamethoxazole resistance was observed in this microorganism (59.0

).