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.

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.

Characterization of trichuris species from porcupine (Hystrix cristata) at zoological garden of Spain.

Population of Trichuris sp. isolated from Hystrix cristata was analyzed based on morphological, biometrical characteristics and mitochondrial (cox1, cob, rrnL) and ribosomal (ITS1) (rDNA) region sequences. Morphological and biometrical results revealed that Trichuris sp. from H. cristata present a high similarity with Trichuris landak from Hystrix javanica and less similarity with other Trichuris species from porcupine species (Trichuris hystricis, Trichuris lenkorani and Trichuris mettami). The lack of molecular data corresponding to Trichuris species that parasitize the porcupine (genus Hystrix and Atelerix) has not allowed a comparative molecular or phylogenetic study. Molecular analyses revealed the existence of two different haplotypes that did not correspond to different morphospecies. Relationships among Trichuris sp. from H. cristata and other Trichuris spp. have been resolved by molecular sequence data in this study. Thus, the combined analysis of one ribosomal and three mitochondrial markers revealed a sister relationship between whipworms parasitizing porcupine and other Trichuris spp. from rodents and canids and separated from the rest of Trichuris spp. from other hosts species. It is necessary accurate information on the possible zoonotic behavior of different Trichuris species for health workers to improve existing control measures. Thus, it is necessary to increase the studies of integrative taxonomy on Trichuris spp. based on morphological, biometrical and molecular data, which will inevitably contribute to our knowledge on the etiology of trichuriasis.

Bacterial communities from Trichuris spp. A contribution to deciphering the role of parasitic nematodes as vector of pathogens.

Microbiome taxa associated with parasitic nematodes is unknown. These invertebrate parasites could act not only as reservoirs and vectors for horizontally transferred virulence factors, but could also provide a potential pool of future emerging pathogens. Trichuris trichiura and Trichuris suis are geohelminths parasitizing the caecum of primates, including humans, and pigs, respectively. The present work is a preliminary study to evaluate the bacterial communities associated with T. trichiura and T. suis, using High Throughput Sequencing and checking the possible presence of pathogens in these nematodes, to determine whether parasitic helminths act as vectors for bacterial pathogens in human and animal hosts. Five T. trichiura adult specimens were obtained from the caecum of macaque (Macaca sylvanus) and two T. suis adults were collected from the caecum of swine (Sus scrofa domestica). The 16S rRNA gene HTS approach was employed to investigate the composition and diversity of bacterial communities in Trichuris spp., with special emphasis at its intestinal level. All samples showed a rich colonization by bacteria, included, preferently, in the phyla Proteobacteria, Firmicutes, Actinobacteria, Bacteroidetes, Cyanobacteria and Verrucomicrobia. A total of 36 phyla and more than 200 families were identified in the samples. Potential pathogen bacteria were detected in these helminths related to the genera Bartonella, Mycobacterium, Rickettsia, Salmonella, Escherichia/Shigella, Aeromonas and Clostridium. The presence of pathogenic bacteria in Trichuris spp. would position these species as a new threat to humans since these nematodes could spread new diseases. This study will also contribute to the understanding of the host-microbiota relation.

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.

Differentiation of Trichuris species eggs from non-human primates by geometric morphometric analysis.

Human trichuriasis is a neglected tropical disease which affects millions of people worldwide, mostly living in low socio-economic conditions. Numerous studies have been conducted over the past 10 years to compare the different techniques for T. trichiura eggs detection. Our study provides the first geometric morphometric analysis for the specific detection of eggs of Trichuris sp. isolated from stools of macaque (M. sylvanus), colobus (C. g. kikuyensis), grivets (C. aethiops) and the Brazza's monkey (C. neglectus) from zoos in Spain. Principal Component Analysis (PCA) arises as an efficient method to determine Trichuris spp. eggs. The selected measurements to be included in the PCA were proposed for the first time in the present work, as far as we know, as we could not find previous studies reporting standardized parameters.

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.