Survival effects of antibiotic exposure during the larval and adult stages in the West Nile virus vector Culex pipiens

The ability of mosquitoes to transmit a pathogen is affected, among other factors, by their survival rate, which is partly modulated by their microbiota. Mosquito microbiota is acquired during the larval phase and modified during their development and adult feeding behavior, being highly dependent on environmental factors. Pharmaceutical residues including antibiotics are widespread pollutants potentially being present in mosquito breeding waters likely affecting their microbiota. Here, we used Culex pipiens mosquitoes to assess the impact of antibiotic exposure during the larval and adult stages on the survival rate of adult mosquitoes. Wild-collected larvae were randomly assigned to two treatments: larvae maintained in water supplemented with antibiotics and control larvae. Emerged adults were subsequently assigned to each of two treatments, fed with sugar solution with antibiotics and fed only with sugar solution (controls). Larval exposure to antibiotics significantly increased the survival rate of adult females that received a control diet. In addition, the effect of adult exposure to antibiotics on the survival rate of both male and female mosquitoes depended on the number of days that larvae fed ad libitum in the laboratory before emergence. In particular, shorter larval ad libitum feeding periods reduced the survival rate of antibiotic-treated adult mosquitoes compared with those that emerged after a longer larval feeding period. These differences were not found in control adult mosquitoes. Our results extend the current understanding of the impact of antibiotic exposure of mosquitoes on a key component of vectorial capacity, that is the vector survival rate.MCIN/AEI/10.13039/501100011033 [grant number PID2020-118205GB-I00]Spanish Ministry of Science and Innovation [grant numbers PRE2021-098544 and FJC2021-048057-I]Spanish Ministry of Universities [Margarita Salas and María Zambrano programs]Funding for open access charge: Universidad de Granada / CBU

Molecular diagnosis of cutaneous leishmaniasis and identification of the causative Leishmania species in Morocco by using three PCR-based assays

Background: The diagnosis of cutaneous leishmaniasis (CL) might be difficult, in particular in endemic areas where different species of Leishmania can cause lesions of very similar appearance and where other skin diseases with similar clinical symptoms occur. Even today, the parasitological diagnosis of CL remains the gold standard and it is based on the direct identification of amastigotes in microscopy smears and/or culture of promastigotes from infected tissues. Although these techniques are highly specific, they are not sensitive enough. The objective of this study is to contribute to improving the diagnosis of CL and the identification of Leishmania species in Morocco by comparing three PCR-based assays applied directly on dermal samples. Methods: A total of 58 patients presenting with cutaneous lesions suggestive of CL were sampled for parasitological diagnosis by direct examination (DE), culture in NNN medium, two kinetoplast DNA (kDNA) PCRs (Lmj4/Uni21 and 13A/13B primers) and one rRNA gene internal transcribed spacer 1 (ITS1) PCR (LITSR/L5.8S primers). The techniques were statistically analyzed and compared. Results: According to our consensus positive, 44 out of 58 samples were true positives. The 13A/13B-PCR and ITS1-PCR showed the highest sensitivities (100%). Parasite microscopy and culture detected 43% and 29% of the true positives, respectively, while culture and microscopy together improved sensitivity to 52%. PCRs 13A/13B and ITS1 were associated to four and one false positives, respectively, while the other assays were 100% specific. Furthermore, the ITS1-PCR-RFLP assay clearly identified the Leishmania species for all the true positives (44/44), whereas Lmj4/Uni21-PCR identified 35/44 samples. The comparison between the Leishmania molecular characterizations and the expected species according to the national data from the Ministry of Health indicate 7 discrepant results. Conclusions: The PCR-based assays tested on our samples increased the speed and sensitivity of the diagnosis of CL compared to the conventional techniques. Furthermore, we showed that we can not base the species identification on the national data from the Ministry of Health. Finally, we suggest the use of PCR-ITS1-RFLP for diagnosis and simultaneous identification of the species in the Moroccan epidemiological context, but also in similar areas of the Mediterranean Basin.This study was supported by the University of Granada, Spain (CICODE program), the Faculty of Medicine and Pharmacy of Casablanca (Morocco), and the PHC Volubilis program

Aedes albopictus in a recently invaded area in Spain: effects of trap type, locality, and season on mosquito captures

Mosquitoes are primary vectors of pathogens impacting humans, wildlife, and livestock. Among them, the Asian tiger mosquito, Aedes albopictus, stands out as an invasive species with a global distribution, having established populations on every continent except Antarctica. Recent findings incriminate Ae. albopictus in the local transmission of several pathogens causing human diseases, including dengue, chikungunya, and Zika viruses and worm parasites as Dirofilaria. In Spain, the establishment of Ae. albopictus occurred in 2004 and it rapidly expanded, currently reaching southern provinces and creating novel epidemiological scenarios in recently invaded areas. In this study, we conducted captures of Ae. albopictus from May to November 2022 in two provinces, Granada and Malaga, situated near the current edge of the species' expanding range in Spain. The objective was to identify the primary factors influencing their captures in these regions. Mosquitoes were captured using BG-Sentinel traps baited with CO2 and BG-Lure, and miniature CDC-UV traps in five different localities. Our findings underscore the influence of both extrinsic factors, such as locality, and intrinsic factors, including mosquito sex, on the abundance of captured Ae. albopictus. A higher abundance of Ae. albopictus was observed in the Malaga province compared to localities in the Granada province. Furthermore, similar numbers of Ae. albopictus mosquitoes were captured in more urbanized areas of Granada, while the lowest counts were recorded in the less urbanized area. These results were compared to captures of another common species in the area, specifically Culex pipiens. Overall, these results represent the first monitoring of invasive Ae. albopictus in the area and are discussed in the light of the potential importance of the species as a nuisance for humans and vectors of pathogens of public health relevance.This study was financed by the PID2020-118205GB-I00 grant to JMP funded by MCIN/AEI/https://doi.org/10.13039/501100011033. Additional support derived from the CNS2022-135993 grant of the Ministerio de Ciencia e Innovación (MCIN/AEI/https://doi.org/10.13039/501100011033) with funding from European Union NextGenerationEU. Mario Garrido was supported by the María Zambrano program and the P9 program for the Incorporation of Young Doctors funded by Spanish Ministry of Universities, the European Union-NextGenerationEU, and the University of Granada. Jesús Veiga received financial support from the Margarita Salas and Juan de la Cierva (FJC2021-048057-I) programs funded by Spanish Ministry of Universities, the Spanish Ministry of Science and Innovation and the European Union-NextGenerationEU. Marta Garrigós was supported by a FPI grant (PRE2021-098544). Mario Garrido is currently granted by the PID2022-137746NA-I00 funded by Spanish Ministry of Science and Innovation. We greatly appreciate the support given by the “Grupo de Investigación Comportamiento y Ecología Animal” of the University of Granada for the field sampling

Canine Leishmaniasis in Southeastern Spain

To examine prevalence changes and risk factors for canine leishmaniasis, we conducted a cross-sectional seroprevalence study and a survey during April–June 2006. Seroprevalence had increased at the meso-Mediterranean bioclimatic level over 22 years. Risk was highest for dogs that were older, large, lived outside, and lived at the meso-Mediterranean level

Publicación en acceso abierto en las universidades andaluzas

Elaborado por el Grupo de Trabajo de Apoyo a la Investigación del CBU

Effectiveness of Fosfomycin for the Treatment of Multidrug-Resistant Escherichia coli Bacteremic Urinary Tract Infections

IMPORTANCE The consumption of broad-spectrum drugs has increased as a consequence of the spread of multidrug-resistant (MDR) Escherichia coli. Finding alternatives for these infections is critical, for which some neglected drugs may be an option. OBJECTIVE To determine whether fosfomycin is noninferior to ceftriaxone or meropenem in the targeted treatment of bacteremic urinary tract infections (bUTIs) due to MDR E coli. DESIGN, SETTING, AND PARTICIPANTS This multicenter, randomized, pragmatic, open clinical trial was conducted at 22 Spanish hospitals from June 2014 to December 2018. Eligible participants were adult patients with bacteremic urinary tract infections due to MDR E coli; 161 of 1578 screened patients were randomized and followed up for 60 days. Data were analyzed in May 2021. INTERVENTIONS Patients were randomized 1 to 1 to receive intravenous fosfomycin disodium at 4 g every 6 hours (70 participants) or a comparator (ceftriaxone or meropenem if resistant; 73 participants) with the option to switch to oral fosfomycin trometamol for the fosfomycin group or an active oral drug or pa renteral ertapenem for the comparator group after 4 days. MAIN OUTCOMES AND MEASURES The primary outcome was clinical and microbiological cure (CMC) 5 to 7 days after finalization of treatment; a noninferiority margin of 7% was considered. RESULTS Among 143 patients in the modified intention-to-treat population (median [IQR] age, 72 [62-81] years; 73 [51.0%] women), 48 of 70 patients (68.6%) treated with fosfomycin and 57 of 73 patients (78.1%) treated with comparators reached CMC (risk difference, -9.4 percentage points; 1-sided 95% CI, -21.5 to infinity percentage points; P = .10). While clinical or microbiological failure occurred among 10 patients (14.3%) treated with fosfomycin and 14 patients (19.7%) treated with comparators (risk difference, -5.4 percentage points; 1-sided 95% CI. -infinity to 4.9; percentage points; P = .19), an increased rate of adverse event-related discontinuations occurred with fosfomycin vs comparators (6 discontinuations [8.5%] vs 0 discontinuations; P = .006). In an exploratory analysis among a subset of 38 patients who underwent rectal colonization studies, patients treated with fosfomycin acquired a new ceftriaxone-resistant or meropenem-resistant gram-negative bacteria at a decreased rate compared with patients treated with comparators (0 of 21 patients vs 4 of 17 patients [23.5%]; 1-sided P = .01). CONCLUSIONS AND RELEVANCE This study found that fosfomycin did not demonstrate noninferiority to comparators as targeted treatment of bUTI from MDR E coli; this was due to an increased rate of adverse event-related discontinuations. This finding suggests that fosfomycin may be considered for selected patients with these infections

Anisakis Infection in the Spotted Flounder Citharus linguatula (Pleuronectiformes: Citharidae) Caught in the Gulf of Cadiz (Area FAO 27-ICES IXa) Appears to Negatively Affect Fish Growth

Spotted flounder (Citharus linguatula L.) caught in the Gulf of Cadiz (area FAO 27 ICES IXa) were examined for Anisakis larvae and to assess the possible risk of anisakiasis in humans through consumption of this fish. Larvae of the genera Anisakis and Hysterothylacium were identified in the analysis of 128 purchased fish specimens. All Anisakis larvae corresponded to type I. Molecular analysis showed the presence of A. pegreffii, A. simplex s.s., and recombinant genotype between the two. The prevalence of Anisakis was 9.4% with a mean intensity of 1.42, while for Hysterothylacium the values were 12.5% and 1.06. The length and weight of the fish, but not Fulton’s condition factor, varied significantly between infected and uninfected fish. The prevalence of Anisakis increased with fish length, with no fish parasitized with Anisakis measuring less than 15.5 cm (2–2.5 years old), which is probably related to the reported dietary change of these fish at around 2 years of age. Fish not parasitized with any of these nematodes showed positive allometric growth, while those parasitized only with Anisakis showed negative allometric growth. When comparing both groups including only fish ≥ 15.5 cm (the smallest size of Anisakis-infected fish), the difference is shown to be statistically significant (p = 0.01), suggesting that Anisakis infection of spotted flounder negatively affects fish growth even when parasite intensity is low, which may have important economic repercussions. Finally, the low prevalence and, above all, intensity of Anisakis in these fish, as well as the habit of consuming this fish fried in oil in our geographical area, means that the risk of acquiring anisakiasis through consumption of this fish is low

Canine Leishmaniasis: Update on Epidemiology, Diagnosis, Treatment, and Prevention

Dog are the main reservoir of Leishmania infantum, causing canine leishmaniasis, an incurable multisystemic disease that leads to death in symptomatic dogs, when not treated. This parasite causes visceral, cutaneous, and mucosal leishmaniasis in people in the Mediterranean Basin, North Africa, South America, and West Asia. This disease is mostly unknown by veterinarians outside the endemic areas, but the disease is expanding in the Northern Hemisphere due to travel and climate change. New methodologies to study the epidemiology of the disease have found new hosts of leishmaniasis and drawn a completely new picture of the parasite biological cycle. Canine leishmaniasis diagnosis has evolved over the years through the analysis of new samples using novel molecular techniques. Given the neglected nature of leishmaniasis, progress in drug discovery is slow, and the few drugs that reach clinical stages in humans are unlikely to be commercialised for dogs, but several approaches have been developed to support chemotherapy. New-generation vaccines developed during the last decade are now widely used, along with novel prevention strategies. The implications of the epidemiology, diagnosis, treatment, and prevention of canine leishmaniasis are fundamental to public health

La leishmaniosis canina en el sudeste de España: cambios en las últimas dos décadas, factores de riesgo y criterios diagnósticos

Las leishmaniosis son parasitosis producidas por protozoos flagelados pertenecientes al género Leishmania (Grande Tejada A.M. et al, 2004). Dicha enfermedad está producida por al menos 20 especies de Leishmania, que son transmitidas por aproximadamente 30 especies de dípteros nematóceros conocidos como flebotomos (vectores probados). Se trata de una parasitosis endémica en 88 países, de los cuales 72 se encuentran en vías de desarrollo. Según la Organización Mundial de la Salud presenta una prevalencia de 12 millones de casos, con una población en riesgo de contraer la enfermedad estimada en 350 millones de personas (Desjeux P., 1996; Desjeux P., 2004). Los estudios realizados en la cuenca mediterránea muestran una fuerte asociación a personas infectadas por VIH, areas rurales y suburbanas, así como una especial prevalencia en grupos de población concernientes al ámbito de la drogodependencia (Pineda J.A. et al., 1998). En España la especie causante de la leishmaniosis es Leishmania infantum, afectando tanto al hombre como al perro (principal reservorio para L. infantum ), (Morillas F. et al, 1995; Martín Sánchez J. et al, 1994). Según Reyes Magañas et al. (1988) la prevalencia de leishmaniosis canina se ha estimado en el 10-15%, llegando en algunos lugares al 25% (Acedo Sánchez et al, 1996). No son pocos los autores que, en los últimos años, han pretendido relacionar los efectos del cambio global, tales como modificaciones en las temperaturas, precipitaciones o humedad (entre otras variables) y sus hipotéticas secuelas sobre la biología y ecología de los vectores, hospedadores intermediarios o reservorios naturales de diversas enfermedades infecciosas (Sutherst R.W., 2004; López Vélez R. et al, 2005). Con el objetivo de comprobar si dicho cambio global podría dar lugar a las mencionadas consecuencias se han llevado a cabo una serie de muestreos que implican tanto a los vectores naturales de Leishmania infantum (diversas especies pertenecientes al género Phlebotomus), como a los reservorios caninos de la enfermedad. Los datos obtenidos fueron comparados con los obtenidos en estudios anteriores a fin de verificar los posibles cambios o fluctuaciones en la densidad y abundancia de los vectores de la enfermedad, así como prevalencia de la leishmaniosis canina. Los mencionados muestreos se realizaron en varias poblaciones pertenecientes a las provincias de Granada y Almería, de igual forma que se hizo en los estudios de referencia para poder comparar los datos empleando los mismos criterios. Finalmente se cotejaron diversas técnicas de diagnóstico de leishmaniosis canina sobre las muestras obtenidas anteriormente (IFI, PCR-ELISA específica de Leishmania infantum y PCR en tiempo real cuantitativa, así como dos técnicas serológicas de diagnóstico rápido basadas en las proteínas rK39 y quimérica recombinante Q).Tesis Univ. Granada. Departamento de Parasitologí