Differentiation of Synanthropic Fleas from Andalusia (Spain) through Geometric Morphometrics Analysis.

Fleas (Siphonaptera) are ectoparasitic hematophagous insects responsible for causing bites and itchy skin conditions in both humans and animals. Furthermore, they can act as vectors of different pathogens of a wide variety of diseases worldwide, including bartonellosis, rickettsiosis, and bubonic plague. Accurate identification of fleas is necessary for the study of their epidemiology, prevention, and control. In addition to traditional morphological classification approaches and molecular biology techniques, geometric morphometrics is increasingly proving to be a useful complementary tool for discriminating between Siphonaptera taxa. With the objective of determining the capacity of this technique to identify and differentiate synanthropic fleas, a principal component analysis was carried out on populations of Ctenocephalides felis, Pulex irritans, and Archaeopsylla erinacei collected in distinct regions of Andalusia (Spain). The analysis carried out on 81 male and female specimens revealed factorial maps that allowed the differentiation of the populations under study, with only partial overlaps that did not prevent their correct identification. Global size differences were also detected, with a slightly larger size in P. irritans males and a bigger size in A. erinacei females. Therefore, the present study emphasizes the role of geometric morphometrics as a useful complementary technique in taxonomic studies of arthropods, especially in the case of flea specimens lacking representative morphological features.

New records of pathogenic bacteria in different species of fleas collected from domestic and peridomestic animals in Spain. A potential zoonotic threat?

Climate change is causing many vectors of infectious diseases to expand their geographic distribution as well as the pathogens they transmit are also conditioned by temperature for their multiplication. Within this context, it is worth highlighting the significant role that fleas can play as vectors of important pathogenic bacteria. For this purpose, our efforts focused on detecting and identifying a total of 9 bacterial genera (Rickettsia sp.; Bartonella sp.; Yersinia sp.; Wolbachia sp., Mycobacterium sp., Leishmania sp., Borrelia sp., Francisella sp. and Coxiella sp.) within fleas isolated from domestic and peridomestic animals in the southwestern region of Spain (Andalusia). Over a 19-months period, we obtained flea samples from dogs, cats and hedgehogs. A total of 812 fleas was collected for this study. Five different species were morphologically identified, including C. felis, C. canis, S. cuniculi, P. irritans, and A. erinacei. Wolbachia sp. was detected in all five species identified in our study which a total prevalence of 86%. Within Rickettsia genus, two different species, R. felis and R. asembonensis were mainly identified in C. felis and A. erinacei, respectively. On the other hand, our results revealed a total of 131 fleas testing positive for the presence of Bartonella sp., representing a prevalence rate of 16% for this genus identifying two species B. henselae and B. clarridgeiae. Lastly, both Y. pestis and L. infantum were detected in DNA of P. irritans and C. felis, respectively isolated from dogs. With these data we update the list of bacterial zoonotic agents found in fleas in Spain, emphasizing the need to continue conducting future experimental studies to assess and confirm the potential vectorial role of certain synanthropic fleas.

Survey of flea infestation in cats and dogs in Western Andalusia, Spain: Seasonality and other risk factors for flea infestation.

This epidemiological survey aims to provide an update on the main flea species that parasitize domestic animals in the Western Andalusia assessing several ecological features that could be considered as possible risk factors for flea infestation. Over a 19-month period (June 2021 to January 2023), we obtained a total of 802 flea samples from 182 dogs (Canis lupus familiaris, Carnivora: Canidae, Linnaeus, 1758) and 78 cats (Felis silvestris catus, Carnivora: Felidae, Schreber, 1775). For each parasitized host, an epidemiological survey was completed, including the following information: geographical origin, age, sex, rural or urban habitat, type of animal's lifestyle (domestic or non-domestic), health status, cohabiting or not with other animals and the total number of collected fleas. The most common species was Ctenocephalides felis (Siphonaptera: Pulicidae) (Bouché, 1835) with a total of 713 specimens, which accounted for 89% of the total fleas. The second most abundant species was Pulex irritans (Siphonaptera: Pulicidae) (Linnaeus, 1758) with a total of 46 collected fleas (6% of the total). The remaining species identified were Archaeopsylla erinacei (Siphonaptera: Pulicidae) (Bouché, 1835) (25 specimens), Spilopsyllus cuniculi (Siphonaptera: Pulicidae) (Dale, 1878) (12 specimens) and Ctenocephalides canis (Siphonaptera: Pulicidae) (Curtis, 1826) (6 specimens), which accounted for 3%, 1% and 1%, respectively, of the total fleas collected. The months with the highest number of collected fleas were, in ascending order, May 2022, September 2021 and July 2021. Dogs had a greater diversity of flea species, and flea sex ratios were female biased in all identified species and among all studied hosts. Finally, we identified some potential host risk factors that promoted higher flea intensities, such as living in rural areas, or presenting other pathologies.

Morphometrics as a Complementary Tool in the Differentiation of Two Cosmopolitan Flea Species: Ctenocephalides felis and Ctenocephalides canis.

Fleas (Siphonaptera) are one of the most important ectoparasites that represent a potential danger for the transmission of pathogens in our environment. The cat flea, Ctenocephalides felis (Bouché, 1835), and the dog flea, Ctenocephalides canis (Curtis, 1826) are among the most prevalent and most frequently studied species throughout the world. However, the variations observed in their morphological characteristics complicate their correct identification, especially when there is a lack of access to the equipment and funds required to carry out molecular biology techniques. With the objective to provide an additional tool to help in the differentiation of Ctenocephalides species, a principal component analysis was carried out for the first time in the present work on populations of C. felis and C. canis from countries in three continents, namely Spain (Europe), South Africa (Africa) and Iran (Asia). The factor maps assisted in the differentiation of both species and the detection of differences in overall size, although morphological ambiguity prevented the delimitation in populations of the same species. Thus, morphometrics represents a complementary tool to other traditional and modern techniques, with great potential to assist in the differentiation of fleas, particularly species that have historically been difficult to identify.

The Use of MALDI-TOF MS as a Diagnostic Tool for Adult Trichuris Species.

Trichuriasis is considered a neglected tropical disease, being the second most common helminthiasis in humans. Detection of Trichuris in routine diagnosis is usually done by microscopic detection of eggs in fecal samples. Other molecular analyses are more reliable and could be used, but these analyses are not routinely available in clinical microbiology laboratories. The use of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MS) is increasing since the last decades due to its recent evidence as a potential role for reliable identification of microorganisms and a few nematodes. But, for parasites detection, normalized protocols and the acquisition and introduction of new species to the database are required. We carried out a preliminary study confirming the usefulness of MALDI-TOF MS for the rapid and reliable identification of Trichuris suis used as control and the creation of an internal database. To create main spectra profiles (MSPs), the different parts of five whipworms (esophagus and intestine) were used, developing different tests to verify the repeatability and reproducibility of the spectra. Thus, to validate the new internal database, 20 whipworms, separating the esophagus and intestine, were used, of which 100% were accurately identified as T. suis, but could not distinguish between both parts of the worm. Log score values ranged between 1.84 and 2.36, meaning a high-quality identification. The results confirmed that MALDI-TOF MS was able to identify Trichuris species. Additionally, a MALDI-TOF MS profile of T. suis proteome was carried out to develop the first internal database of spectra for the diagnosis of trichuriasis and other Trichuris spp.

Comparative molecular and morphological study of Stenoponia tripectinata tripectinata (Siphonaptera: Stenoponiidae) from the Canary Islands and Corsica.

Stenoponia tripectinata tripectinata (Tiraboschi, 1902) is the most prevalent subspecies, within the genus Stenoponia, in the Mediterranean area. This rodent flea is widely distributed throughout southwestern Europe and the North of Africa including Mediterranean islands and the Canary Islands. Nevertheless, from a taxonomical and systematic point, this flea group has been neglected over the years. Therefore, the aim of this study was to carry out a comparative morphometric, phylogenetic, and molecular study of two populations of S. t. tripectinata isolated from rodents collected from different islands from the Canary Archipelago and from Corsica to clarify the taxonomic status of these two isolated populations and to assess the morphological and molecular differentiation between them. For this purpose, we have analyzed several morphological traits and sequenced five molecular markers (EF1-α, ITS2, cox1, cox2, and cytb). We observed slight differences in the overall body size between females of both populations, and two well-defined geographical genetic lineages. This suggests the existence of two cryptic subspecies within S. t. tripectinata corresponding to two different island groups. Furthermore, we bring to light the necessity to provide new and updated morphological, molecular, and phylogenetic data to clarify the taxonomic status of S. tripectinata.

New records of bacteria in different species of fleas from France and Spain.

In this study, we assessed the presence of vector-borne microorganisms in different species of fleas collected from different hosts in diverse areas of South-Western Europe by molecular methods. A total of 319 fleas belonging to eight different species was tested for the presence of eight microorganisms. Wolbachia spp. endosymbionts were detected in Ctenocephalides felis, Pulex irritans, Archaeopsylla erinacei and Ctenophthalmus baeticus boisseauorum specimens. Rickettsia felis, an emerging pathogen, was detected in C. felis, A. erinacei and Ct. b. boisseauorum. Rickettsia typhi, the agent of murine typhus was detected for the first time in A. erinacei and Mycobacterium spp. were detected for the first time in fleas (C. felis, P. irritans and A. erinacei). Lastly, five different species of Bartonella were detected in fleas' DNA in this study, including a possible new bacterium belonging to this genus. With this study, we updated the knowledge of the flea-borne bacteria present in the South-West of Europe reinforcing the idea about the necessity to expand and increase the current knowledge on flea-borne pathogens.

Trichuris trichiura isolated from Macaca sylvanus: morphological, biometrical, and molecular study.

BACKGROUND: Recent studies have reported the existence of a Trichuris species complex parasitizing primate. Nevertheless, the genetic and evolutionary relationship between Trichuris spp. parasitizing humans and Non-Human Primates (NHP) is poorly understood. The hypothesised existence of different species of Trichuris in primates opens the possibility to evaluate these primates as reservoir hosts of human trichuriasis and other putative new species of whipworms. RESULTS: In this paper, we carried out a morphological, biometrical and molecular study of Trichuris population parasitizing Macaca sylvanus from Spain based on traditional morpho-biometrical methods, PCA analysis and ribosomal (ITS2) and mitochondrial (cox1 and cob) DNA sequencing. Morphological results revealed that Trichuris sp. from M. sylvanus is Trichuris trichiura. Ribosomal datasets revealed that phylogenetic relationships of populations of Trichuris sp. from M. sylvanus were unresolved. The phylogeny inferred on mitochondrial datasets (partitioned and concatenated) revealed similar topologies; Thus, phylogenetic trees supported the existence of clear molecular differentiation between individuals of Trichuris sp. from M. sylvanus appearing in two different subclades. CONCLUSIONS: Based on morphological parameters, biometrical measurements, and molecular sequence analysis, we conclude that the whipworms isolated from M. sylvanus were T. trichiura. Further, the evolutionary relationship showed that these worms belonged to two genotypes within the T. trichiura lineage. Since T. trichiura is of public health importance, it is important to carry out further studies to improve the understanding of its hosts range, evolution and phylogeography.

Fleas and flea-borne diseases of North Africa.

North Africa has an interesting and rich wildlife including hematophagous arthropods, and specifically fleas, which constitute a large part of the North African fauna, and are recognised vectors of several zoonotic bacteria. Flea-borne organisms are widely distributed throughout the world in endemic disease foci, where components of the enzootic cycle are present. Furthermore, flea-borne diseases could re-emerge in epidemic form because of changes in the vector-host ecology due to environmental and human behaviour modifications. We need to know the real incidences of flea-borne diseases in the world due to this incidence could be much greater than are generally recognized by physicians and health authorities. As a result, diagnosis and treatment are often delayed by health care professionals who are unaware of the presence of these infections and thus do not take them into consideration when attempting to determine the cause of a patient's illness. In this context, this bibliographic review aims to summarise the main species of fleas present in North Africa, their geographical distribution, flea-borne diseases, and their possible re-emergence.

Ctenophthalmus baeticus boisseauorum (Beaucournu, 1968) and Ctenophthalmus apertus allani (Smit, 1955) (Siphonaptera: Ctenophthalmidae) as synonymous taxa: morphometric, phylogenetic, and molecular characterization.

The family Ctenophthalmidae (Order Siphonaptera) has been considered as a 'catchall' for a wide range of divergent taxa showing a paraphyletic origin. In turn, Ctenophthalmus sp. (Ctenophthalmidae) includes 300 valid described taxa. Within this genus, males are easily distinguishable basing on the size, shape, and chaetotaxy of their genitalia; however, females show slight morphological differences with each other. The main objective of this work was to carry out a comparative morphometric, phylogenetic, and molecular study of two different subspecies: Ctenophthalmus baeticus boisseauorum and Ctenophthalmus apertus allani in order to clarify and discuss its taxonomic status. From a morphological and biometrical point of view, we found clear differences between modified abdominal segments of males of both subspecies and slight differences in the margin of sternum VII of all female specimens which did not correspond with molecular and phylogenetic results based on four different molecular markers (Internal Transcribed Spacer 1 and 2 of ribosomal DNA, and the partial cytochrome c oxidase subunit 1 and cytochrome b of mitochondrial DNA). Thus, we observed a phenotypic plasticity between both subspecies, which did not correspond with a real genotypic variability nor different environmental or ecological conditions. Basing on these results, we could consider that there are no solid arguments to consider these two 'morphosubspecies' as two different taxa. We propose that C. b. boisseauorum should be considered as a junior synonym of C. a. allani.

Evidence of cryptic species in the genus Tinaminyssus (Acari: Rhinonyssidae) based on morphometrical and molecular data.

The study of cryptic species allows to describe and to understand biodiversity, and the evolutionary processes shaping it. Mites of the family Rhinonyssidae are permanent parasites of the nasal cavities of birds, currently including about 500 described species and 12 genera. Here, we tested the hypothesis that mites from five populations of the genus Tinaminyssus-three isolated from European turtle doves (Streptopelia turtur), and two from Eurasian collared doves (Streptopelia decaocto; Aves: Columbiformes)-are, in fact, two cryptic species inhabiting different hosts. First, we performed a morphometrical study on 16 traits. Then, we used the ITS1-5.8S rDNA-ITS2 nuclear region (ITS region), and a fragment of the mitochondrial cytochrome c-oxidase 1 (COI) to carry out phylogenetic and species delimitation analyses on Tinaminyssus species. Morphological analyses revealed a lack of biometric differentiation among Tinaminyssus populations from the two host species. However, molecular analyses indicated a high degree of genetic differentiation between populations of Tinaminyssus sp. from S. turtur and S. decaocto. Overall, results show that they can be considered as different cryptic species, suggesting a case of evolutionary stasis, likely because of the anatomical similarity between closely-related bird host species.

Infection Rates of Wolbachia sp. and Bartonella sp. in Different Populations of Fleas.

In the present study, a molecular detection of Bartonella sp. and Wolbachia sp. in Ctenocephalides felis (Siphonaptera: Pulicidae) isolated from Canis lupus familiaris from different geographical areas of Spain, Iran and South Africa, and in Stenoponia tripectinata tripectinata isolated from Mus musculus from the Canary Islands has been carried out by amplification of the 16S ribosomal RNA partial gene of Wolbachia sp. and intergenic spacer region (its region) of Bartonella sp. A total of 70 % of C. felis analysed were infected by W. pipientis. This percentage of prevalence was considerably higher in female fleas than in male fleas. Bartonella DNA was not detected in C. felis from dogs, while Bartonella elizabethae was detected and identified in S. t. tripectinata from M. musculus from the Canary Islands representing 43.75 % prevalence. This report is the first to identify B. elizabethae in S. t. tripectinata collected in M. musculus from the Canary Islands. Thus, our results demonstrate that this flea is a potential vector of B. elizabethae and might play roles in human infection. The zoonotic character of this bartonellosis emphasizes the need to alert public health authorities and the veterinary community of the risk of infection.

Trichuris colobae n. sp. (Nematoda: Trichuridae), a new species of Trichuris from Colobus guereza kikuyensis.

In the present work, a morphological and biometrical study of whipworms Trichuris Roederer, 1761 (Nematoda: Trichuridae) parasitizing Colobus guereza kikuyensis has been carried out. Biometrical and statistical data showed that the mean values of individual variables between Trichuris suis and Trichuris sp. from C. g. kikuyensis differed significantly (P < 0.001) when Student's t test was performed: seven male variables (width of esophageal region of body, maximum width of posterior region of body, width in the place of junction of esophagus and the intestine, length of bacillary stripes, length of spicule, length of ejaculatory duct, and distance between posterior part of testis and tail end of body) and three female variables (width of posterior region of body, length of bacillary stripes, and distance of tail end of body and posterior fold of seminal receptacle). The combination of these characters permitted the discrimination of T. suis with respect to Trichuris sp. from C. g. kikuyensis, suggesting a new species of Trichuris. Furthermore, males of Trichuris sp. from C. g. kikuyensis showed a typical subterminal pericloacal papillae associated to a cluster of small papillae that were absent in males of T. suis, while females of Trichuris from Colobus appeared with a vulval region elevated/over-mounted showing a crater-like appearance. The everted vagina showed typical triangular sharp spines by optical microscopy and SEM. Thus, the existence of a new species of Trichuris parasitizing C. g. kikuyensis has been proposed.

Molecular characterization and phylogeny of whipworm nematodes inferred from DNA sequences of cox1 mtDNA and 18S rDNA.

A molecular phylogenetic hypothesis is presented for the genus Trichuris based on sequence data from the mitochondrial cytochrome c oxidase 1 (cox1) and ribosomal 18S genes. The taxa consisted of different described species and several host-associated isolates (undescribed taxa) of Trichuris collected from hosts from Spain. Sequence data from mitochondrial cox1 (partial gene) and nuclear 18S near-complete gene were analyzed by maximum likelihood and Bayesian inference methods, as separate and combined datasets, to evaluate phylogenetic relationships among taxa. Phylogenetic results based on 18S ribosomal DNA (rDNA) were robust for relationships among species; cox1 sequences delimited species and revealed phylogeographic variation, but most relationships among Trichuris species were poorly resolved by mitochondrial sequences. The phylogenetic hypotheses for both genes strongly supported monophyly of Trichuris, and distinct genetic lineages corresponding to described species or nematodes associated with certain hosts were recognized based on cox1 sequences. Phylogenetic reconstructions based on concatenated sequences of the two loci, cox1 (mitochondrial DNA (mtDNA)) and 18S rDNA, were congruent with the overall topology inferred from 18S and previously published results based on internal transcribed spacer sequences. Our results demonstrate that the 18S rDNA and cox1 mtDNA genes provide resolution at different levels, but together resolve relationships among geographic populations and species in the genus Trichuris.

[Health related problems as cause of visits to hospital emergency departments]. / Problemas de salud relacionados con los medicamentos en un servicio de urgencias hospitalario.

BACKGROUND AND OBJECTIVE: Medication-related problems (MRPs) are health problems resulting from patient pharmacotherapy failure that interfere with the expected outcome in the patient's health status. The aim of this study was to discover the health problems associated with the appearance of MRPs. PATIENTS AND METHOD: Patient interviews, together with the assessment of medical records were the sources of information used in the assessment and identification of MRPs. A validated questionnaire was used for the interviews; in order to classify the health problems found, the ICD-9 was used. RESULTS: 2,556 patients were interviewed over a period of one year at a hospital emergency department. 2,261 of these cases were valid. Osteoarticular diseases, poorly defined signs and symptoms of illness, injuries and intoxications were the disorders most commonly associated with the appearance of MRPs. MRPs of necessity and effectiveness had a similar profile. MRPs of safety were more commonly associated with poorly defined signs and symptoms of illness, injuries and intoxications, as well as nervous, digestive and blood problems. CONCLUSIONS: Most frequent medication related health problems causing visits to hospital emergency departments were osteoarticular disorders, followed by poorly defined signs and symptoms of illness. Differences were observed in the diagnoses between the different dimensions and types of MRP.

Problemas de salud relacionados con los medicamentos en un servicio de urgencias hospitalario / Health related problems as cause of visits to hospital emergency departments

FUNDAMENTO Y OBJETIVO: Los problemas relacionados con los medicamentos (PRM) son problemas de salud que se producen como consecuencia de fallos de la farmacoterapia del paciente y que interfieren con los resultados esperados de salud. El objetivo de este trabajo es conocer los problemas de salud más frecuentes que constituyen PRM. PACIENTES Y MÉTODO: La entrevista con el paciente junto con la evaluación de la historia clínica son las principales fuentes de información para evaluar e identificar PRM. Se utilizó un cuestionario validado como instrumento para la entrevista y la CIE-9 para la clasificación de los problemas de salud encontrados. RESULTADOS: Se entrevistó a un total de 2.556 pacientes durante 1 año en el servicio de urgencias hospitalario, de los que se estudiaron 2.261 tras las exclusiones. Las enfermedades osteoarticulares, los signos y síntomas mal definidos y las lesiones y envenenamientos son los problemas más relacionados con la aparición de PRM. Los PRM de necesidad y efectividad mantienen un perfil similar. Los PRM de seguridad se asociaron más a signos y síntomas mal definidos y lesiones y envenenamientos así como a alteraciones nerviosas, digestivas y sanguíneas. CONCLUSIONES: Los PRM más frecuentes que son causa de consulta en un servicio de urgencias hospitalario son los osteoarticulares, seguidos de los signos y síntomas mal definidos. Se observan diferencias en los diagnósticos entre las distintas dimensiones de PRM y sus tipos

BACKGROUND AND OBJECTIVE: Medication-related problems (MRPs) are health problems resulting-from patient pharmacotherapy failure that interfere with the expected outcome in the patient’s health status. The aim of this study was to discover the health problems associated with the appearance of MRPs. PATIENTS AND METHOD: Patient interviews, together with the assessment of medical records were the sources of information used in the assessment and identification of MRPs. A validated questionnaire was used for the interviews; in order to classify the health problems found, the ICD-9 was used. RESULTS: 2,556 patients were interviewed over a period of one year at a hospital emergency department.2,261 of these cases were valid. Osteoarticular diseases, poorly defined signs and symptoms of illness, injuries and intoxications were the disorders most commonly associated with the appearance of MRPs. MRPs of necessity and effectiveness had a similar profile. MRP sof safety were more commonly associated with poorly defined signs and symptoms of illness, injuries and intoxications, as well as nervous, digestive and blood problems. CONCLUSIONS: Most frequent medication related health problems causing visits to hospital emergency departments were osteoarticular disorders, followed by poorly defined signs and symptoms of illness. Differences were observed in the diagnoses between the different dimensions and types of MRP