Influence of egg density on larval development and adult body size of cat fleas (Ctenocephalides felis).

Fleas (Siphonaptera) are holometabolous insects with larval and adult stages that exhibit vastly different ecologies from each other. Adult fleas are parasitic and feed exclusively on the blood of a vertebrate host, whereas flea larvae do not live on hosts and consume dried faecal blood from adult fleas. Because flea larvae rely on adult flea faeces for food, excrement and eggs must fall in the same location; thus, larval density is likely high in these restricted habitats. However, the influence of larval density on the subsequent adult stage has not been examined. In the present study, we utilized egg density to investigate density-dependent effects on larval development and adult body size in the cat flea (Ctenocephalides felis Bouché) (Siphonaptera: Pulicidae). Specifically, eggs were collected to create three different larval densities (n = 50, 100 and 150 per 56.7 cm2), and hatched larvae from all groups were fed an excess amount of adult faecal pellets. Larval development was measured by recording the proportion of eggs that developed to the pupal stage and the proportion of eggs that reached adulthood (eclosion). The body size of eclosed adults was quantified for both sexes using head length and length of the total body. We found that the number of eggs had no effect on the proportion of larvae that pupated or the proportion of larvae that eclosed; however, higher egg densities resulted in larger body sizes for both sexes. Overall, these data yield significant insight into how the ecology of larval fleas impacts the biology of the resultant adults.

Evidence of castrations in fleas of small mammals (Chiroptera and Rodentia) from Argentina.

Morphological abnormalities in fleas seem to be common in nature and are under reported in Argentina. In this note, we describe anomalies in two males and one female of Alectopsylla unisetosa Mahnert (Ischnopsyllidae) and one of Polygenis sp. (Rhopalopsyllidae) male collected from small mammals in the provinces of Neuquen and Salta, Argentina. In all specimens, the anomalies were observed at the level of the genitalia recognized as partial castration. The structures mainly affected were the modified abdominal segments, the aedeagus (in male), and the spermatheca (in female). The present communication is the first one devoted exclusively to teratogenous fleas in Argentina.

Seasonality and other risk factors for fleas infestations in domestic dogs and cats.

Fleas in the genus Ctenocephalides are the most clinically important parasitic arthropods of dogs and cats worldwide yet risk factors that might increase the risk of infestation in small animals remains unclear. Here we developed a supervised text mining approach analysing key aspects of flea epidemiology using electronic health records from domestic cats and dogs seen at a sentinel network of 191 voluntary veterinary practices across Great Britain between March 2014 and July 2020. Our methods identified fleas as likely to have been present during 22,276 of 1,902,016 cat consultations (1.17%) and 12,168 of 4,844,850 dog consultations (0.25%). Multivariable logistic regression modelling found that animals originating from areas of least deprivation were associated with 50% reductions in odds of veterinary-recorded flea infestation compared to the most deprived regions in England. Age of the animal was significantly associated with flea presentation in both cats and dogs, with cases peaking before animals reached 12 months. Cases were recorded through each study years, peaking between July and October, with fluctuations between each year. Our findings can be used towards healthcare messaging for veterinary practitioners and owners.

Host specificity of flea parasites of mammals from the Andean Biogeographic Region.

Host specificity of fleas affects their biodiversity that plays a major role in determining the potential transmission routes by pathogens through vertebrate hosts, including humans. In the Biogeographic Andean region, numerous systematic and ecological studies have been conducted, revealing a high diversity of flea taxa of mammals and the presence of pathogenic organisms transmitted by fleas; however, the degree of preference with which each flea species associates with a mammal host remains poorly understood in this region. Herein, host specificity in mammal fleas from the Andean region was analysed. We employed the number of host species for each flea species and the index of host specificity STD *. Following the literature, 144 species and 13 subspecies of fleas (31 genera and 10 families) have been described in the Andean biogeographic region; 76 taxa are endemic to this region. To carry out the analyses of host specificity, we considered 1759 records of fleas collected from 124 species and 59 genera of wild and domestic mammals, mostly rodent species (85.9%). Our results indicate that typical Andean fleas are genus or family host specific (mostly STD * less than 3.0). More diverse mammal hosts are parasitized by more diverse flea genera and families and these hosts are phylogenetically related. Otherwise, these hosts are associated with different flea lineages, suggesting the interaction of ecological and evolutionary mechanisms (host-switching, ecological adaptations and co-evolutionary alternation). The fields of disease ecology and One Health are considering the host specificity of arthropod vectors as an important point to understand the mechanisms of emergence and re-emergence of diseases. Our results allow us to estimate the risk of diseases involving fleas in the Andean region.

Host body mass, not sex, affects ectoparasite loads in yellow-necked mouse Apodemus flavicollis.

We investigated the presence and potential causes of sex bias in ectoparasite infestations in the yellow-necked mouse Apodemus flavicollis. We compared the natural tick and flea burdens of male and female mice in a temperate beech forest and assessed whether the observed differences were driven by host sex or body mass. We found that males were more heavily infested by ticks compared to female mice. However, this difference was driven by host body mass, and not sex itself. Host body mass positively correlated with flea loads, but there was no evidence of sex bias in flea abundance. In addition, the abundance of both ticks and fleas infesting yellow-necked mice changed over time, both seasonally (month to month) and annually (year to year). Our results underscore the importance of the sexual size dimorphism and the parasite taxon as the primary factors that influence the occurrence of sex-biased parasitism in small mammals.

Culling of Urban Norway Rats and Carriage of Bartonella spp. Bacteria, Vancouver, British Columbia, Canada.

We investigated the effects of culling on Bartonella spp. bacteria carriage among urban rats in Canada. We found that the odds of Bartonella spp. carriage increased across city blocks except those in which culling occurred. Removing rats may have prevented an increase in Bartonella spp. prevalence, potentially lowering human health risks.

Ecology of fleas and their hosts in the trifinio of north-east Argentina: first detection of Rickettsia asembonensis in Ctenocephalides felis felis in Argentina.

Fleas are important in public health due to their role as parasites and vectors of pathogens, including Rickettsia. The aim of this study was to evaluate the diversity, abundance and prevalence of fleas and the presence of Rickettsia in the trifinio of north-east Argentina. Fleas from household and synanthropic animals were obtained from urban and periurban areas. They were taxonomically identified and samples of 227 fleas in 86 pools were analysed by polymerase chain reaction targeting the gltA and ompB genes of Rickettsia spp. The study revealed that Ctenocephalides felis felis was dominant on dogs, cats and opossums, with higher prevalence in the periurban area. The Shannon-Wiener and Morisita-Horn indices expressed differences in the diversity and similarity values of the absolute abundances of the species between the areas compared. DNA amplifications revealed 30.8% C. f. felis pools positive for Rickettsia spp. Phylogenetic analysis showed that the haplotype obtained was identical to Rickettsia asembonensis from Peru and Brazil. This is the first detection in Argentina of R. asembonensis that infects C. f. felis, and we emphasize the importance of conducting research from a 'One Health' perspective on the role of opossums and rodents in the integration of the transmission cycles of rickettsial bacteria.

Immune defense mechanisms against a systemic bacterial infection in the cat flea (Ctenocephalides felis).

A significant amount of work has been devoted towards understanding the cellular and humoral immune responses in arthropod vectors. Although fleas (Siphonaptera) are vectors of numerous bacterial pathogens, few studies have examined how these insects defend themselves from infection. In this study, we investigated the immune defense mechanisms in the hemocoel of cat fleas (Ctenocephalides felis), currently the most important flea pest of humans and many domestic animals. Using model species of bacteria (Micrococcus luteus, Serratia marcescens, and Escherichia coli), we delivered a systemic infection and measured the following: antimicrobial activity of hemolymph, levels of free radicals resulting from the induction of oxidase-based pathways, number of circulating hemocytes, phagocytosis activity of circulating hemocytes, and in vivo bacteria killing efficiency when phagocytosis activity is limited. Our results show that the antimicrobial activity of flea hemolymph increases in response to certain species of bacteria; yet, a systemic infection with the same bacterial species did not influence levels of hydrogen peroxide (H2O2), a reactive intermediate of oxygen, at the same time. Additionally, the number of circulating hemocytes increases in response to E. coli infection, and these cells display strong phagocytic activity against this bacterium. Moreover, limiting phagocytosis by injecting polystyrene beads subsequently increases flea susceptibility to E. coli infection when compared to injury controls; however, impairing the cellular immune response itself did not increase flea susceptibility to infection when compared to untreated fleas. Overall, this work yields significant insight into how fleas interact with bacterial pathogens in their hemocoel, and suggests that cellular and humoral immune responses cooperate to combat systemic bacterial infections.

New taxonomic and evolutionary insights relevant to the cat flea, Ctenocephalides felis: A geographic perspective.

The cat flea, Ctenocephalides felis, is an obligate haematophagous ectoparasite of wildlife and domestic cats and dogs worldwide. Since cat fleas can affect the health of humans and their pets, an uncertain taxonomy of this taxon can greatly inhibit pest and disease management. To address the evolution and taxonomy of the cat flea, we set out to determine 1) how many genetically distinct taxa exist, 2) whether there is morphological support for the genetically distinct taxa, and 3) the role of host range and paleoclimatic events in speciation. We collected a total of 3352 fleas sampled from 576 domestic cats and dogs as well as 10 wildlife species across 30 localities in South Africa. A total of three flea genera, five species, and three of the currently recognized cat flea subspecies, C. f. damarensis, C. f. strongylus and C. f. felis were obtained. Geometric morphometric analyses on head shape were performed on 68 female and 107 male cat flea individuals. Principal component analysis demonstrated large overlap in head shape variation between C. f. strongylus and C. f. felis, rendering this character not useful for phylogenetic inferences. DNA was extracted from 188 Ctenocephalides spp. and mitochondrial COII and nuclear EF1-α sequences were generated. Bayesian and Maximum Likelihood analyses as well as a TCS parsimony haplotype network of the mitochondrial DNA confirmed the presence of three well supported monophyletic clades. These assemblages did not fully corroborate the existence of the three C. felis subspecies. A single well-supported molecular clade included only C. f. damarensis morphotypes that were mostly collected from wildlife. The recognition of this subspecies as a distinct taxon was further corroborated by sequence distances and also the number of plantar spiniform bristles on fore-tarsi V in males. Despite the overall lack of support for the recognition of C. f. damarensis and C. f. strongylus, a geographic trend was visible whereby one genetic lineage corresponded to the western dryer hot subregion, whereas the other was found throughout the region. Bayesian dating suggested that these two clades diverged during the early Pliocene (4.18 mya), a date that corresponds well with the establishment of a dry hot climate in the west of southern Africa. If so, the off-host environment, particularly temperature and humidity, are important factors to consider in the evolution of the cat flea. The present study rejects recent assertions that the three cat flea subspecies are valid entities and rather point to a situation where more sampling is required before the taxonomic status of C. f. damarensis can be resolved.

Diversity, distribution and changes in communities of fleas on small mammals along the elevational gradient from the Pannonian Plain to the Carpathian Mountains.

The causal chain of parasite-host-environment interactions, the so-called 'dual parasite environment', makes studying parasites more complicated than other wild organisms. A sample of 65 282 fleas taken from 336 different locations were analysed for changes in the distribution, diversity and compensation of flea communities found on small mammals along an elevational diversity gradient ranging from the Pannonian Plain to the base of the Carpathian summits. The fleas were divided into four groups, which were derived from changes in abundance and occurrence determined from cluster analysis. They are (1) flea species whose range seems unrelated to any change in elevation; (2) species that avoid high altitudes; (3) a group that can be subdivided into two types: (i) host-specific fleas and (ii) mountains species and (4) species opting for high altitudes on the gradient or preferring lower to middle elevations below 1000 m. Our study showed a unimodal pattern of flea diversity along the elevational gradient. It indicated that seasonality significantly conditions changes in biodiversity and patterns of spatial change along the elevational gradient, with specific flea species influenced by their host, while the impact of environmental conditions is more pronounced in opportunistic flea species.

Detection of Zoonotic Bartonella Pathogens in Rabbit Fleas, Colorado, USA.

We detected 3 Bartonella species in wild rabbit fleas from Colorado, USA: B. vinsonii subsp. berkhoffii (n = 16), B. alsatica (n = 5), and B. rochalimae (n = 1). Our results support the establishment of the zoonotic agent B. alsatica in North America.

Plague Epizootic Dynamics in Chipmunk Fleas, Sierra Nevada Mountains, California, USA, 2013-2015.

We describe Yersinia pestis minimum infection prevalence in fleas collected from Tamias spp. chipmunks in the Sierra Nevadas (California, USA) during 2013-2015. Y. pestis-positive fleas were detected only in 2015 (year of plague epizootic), mostly in T. speciosus chipmunks at high-elevation sites. Plague surveillance should include testing vectors for Y. pestis.

Clinical and pathological aspects of first report of Tunga penetrans infestation on southern brown howler monkey (Alouatta guariba clamitans) in Rio Grande do Sul, Brazil.

BACKGROUND: Tungiasis is a neglected neotropical disease caused by penetration of Tunga spp. into the skin of the host. METHODS: Two primates were rescued from nearby different indigenous villages, and the clinical, pathological, and parasitological features of tungiasis were described. Flea identification occurred through their morphometry and was confirmed with the use of a dichotomous key. RESULTS: Monkey 1 was parasitized by 23 sand fleas and, after treatment, was assigned to the animal rehabilitation center. Monkey 2 was in poor body condition and died shortly after clinical examination. At necropsy, this primate was parasitized by 26 specimens of sand fleas. CONCLUSIONS: Both animals altered their tree behavior by staying on the ground for long periods. This parasitic relationship implies the possibility of enlargement of the sand flea dispersion. Thus, this is the first record of Tunga penetrans occurrence in wild Alouatta guariba clamitans.

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.

Molecular detection of Bartonella in fleas (Hexapoda, Siphonaptera) collected from wild rodents (Cricetidae, Sigmodontinae) from Argentina.

Bartonella are facultative intracellular Gram-negative bacteria, transmitted mainly by hematophagous arthropods, and the rodents act as a natural reservoir. Different species of Bartonella associated with rodents have been implicated as causing human disease. Studies from Argentina are scarce and no Bartonella from fleas have been reported previously. The present study investigated the presence of Bartonella spp. in fleas associated with sigmodontine rodents in four localities of the Santa Cruz Province, Argentina. In total, 51 fleas (four species) were analysed of which 41.2% were found to be positive for the gltA gene fragment via a nested polymerase chain reaction. All positive fleas were of the species Neotyphloceras crackensis from three different localities. Eight of the 21 amplified samples were sequenced, and the presence of three different genotypes was detected with an identity of 95.5-98.8% amongst themselves. Bartonella genotypes from American rodents and rodent fleas were recovered in a monophyletic group. Similarly, most of the Peruvian and all Argentinean variants constitute a natural group sister of the American remainder. The importance of the Bartonella spp. with respect to public health is unknown, although future studies could provide evidence of the possible involvement of N. crackensis in the Bartonella transmission cycles.

Origin, evolution, phylogeny and taxonomy of Pulex irritans.

The human flea Pulex irritans Linnaeus, 1758 (Siphonaptera: Pulicidae) is one of the most studied species together with the cat flea Ctenocephalides felis Bouché, 1835, because they have a cosmopolitan distribution and are closely related to humans. The present study aimed to carry out a comparative morphometric and molecular study of two different populations of P. irritans (Spain and Argentina). Accordingly, internal transcribed spacer (ITS)1 and ITS2 of rDNA and the partial cytochrome c oxidase subunit 1 (cox1) and cytochrome b (cytb) mtDNA genes of these taxa were sequenced. Furthermore, the taxonomy, origin, evolution and phylogeny of P. irritans was assessed. The morphometric data obtained did not show significant differences between P. irritans specimens from Spain and Argentina, even when these two populations were collected from different hosts; however, there was a considerable degree of molecular divergence between both populations based on nuclear and mitochondrial markers. Thus, it is proposed that P. irritans, in contrast with other generalist fleas, maintains a certain degree of morphological similarity, at least between Western Palearctic and Neotropical areas. Furthermore, two well defined geographical genetic lineages within the P. irritans species are indicated, suggesting the existence of two cryptic species that could be discriminated by a polymerase chain reaction-linked restriction fragment length polymorphism.

Diversity, distribution and parasitism rates of fleas (Insecta: Siphonaptera) on sigmodontine rodents (Cricetidae) from Argentinian Patagonia.

Fleas have great medical relevance as vectors of the causative agents of several diseases in animals and humans and rodents are the principal reservoirs for these pathogens. Argentinian Patagonia has the highest diversity of rodent fleas in South America. However, parasitism rates of rodents by fleas, the factors that influence them and the ecological aspects that modulate geographical distributions of flea-host association remain unknown for this region. This is the first study to record the diversity, prevalence, abundance, geographical distributions and host ranges of fleas in Argentinian Patagonia. It also compares parasitism rates among Patagonian ecoregions and host species. We captured 438 rodents belonging to 13 species, which harboured 624 fleas from 11 species and subspecies (P = 46%; mean abundance = 1.44). The high parasitism rates obtained were consistent with previous records for other arid regions, suggesting that Patagonia favours the survival and development of Siphonaptera. Host geographic range and abundance were related to the parasitological indexes: host species with high-density populations had the highest mean flea abundance and prevalence, whereas widely distributed hosts had the highest richness and diversity of flea species. Our results contribute to the knowledge of the flea-host-environment complex. Our analysis of flea distributions and parasitism rate in Central Patagonia may be useful in epidemiological studies of flea-borne diseases and provide a basis for implementing surveillance systems for better risk assessment of emerging zoonoses in the region.

Fleas of the genus Neotyphloceras associated with rodents from Bolivia: new host and distributional records, description of a new species and remarks on the morphology of Neotyphloceras rosenbergi.

The flea genus Neotyphloceras Rothschild (Siphonaptera: Ctenophthalmidae) includes five species and two subspecies distributed from Venezuela to southern Chile and Argentina. Only Neotyphloceras crassispina hemisus Jordan has been registered in Bolivia. The present study examines species of Neotyphloceras collected in Bolivian localities in the Departments of La Paz, Cochabamba and Tarija, and describes the morphology of the modified abdominal segments in males and females of Neotyphloceras rosenbergi Rothschild on the basis of type material and specimens collected from Tarija. A new species, Neotyphloceras boliviensis n. sp., is described and new host associations are reported for N. rosenbergi, Neotyphloceras crassispina crassispina and N. crassispina hemisus. Neotyphloceras c. crassispina and N. rosenbergi are reported for the first time in Bolivia. The distribution of N. rosenbergi is extended 1600 km to the south. Given the potential medical and veterinary significance of fleas as disease vectors, and considering that in the Departments of La Paz and Tarija several human cases of plague have been reported, and species of flea have been identified as main vectors of these diseases, the new records of fleas in Bolivia reported herein may be useful for epidemiological studies on flea-borne diseases.

Descriptions of two new species of flea of the genus Plocopsylla in northwestern Argentina.

Two new species of flea of the genus Plocopsylla, subgenus Plocopsylla, (Siphonaptera: Stephanocircidae) collected from sigmodontine rodents in northwestern Argentina are described and a key to identification of species of the genus Plocopsylla, subgenus Plocopsylla, in Argentina is presented. Plocopsylla (P.) inti is cited for the first time in Argentina, extending its distribution ∼ 970 km further south than previously documented. New locality data and flea-host associations are recorded. The contributions of this study are relevant because they increase knowledge of the diversity of flea fauna in northwestern Argentina bordering a plague endemic area and will be useful in the implementation of plague control management plans in the future.

Morphological asymmetry and habitat quality: using fleas and their rodent hosts as a novel experimental system.

Morphological asymmetry is widely used to measure developmental instability and higher levels of asymmetry often correlate with decreased mating success, increased inbreeding, increased stress and decreased habitat quality. We studied asymmetry and relationships between asymmetry and host identity in two flea species, host generalist Xenopsylla ramesis and host specialist Parapulex chephrenis, and asked: (1) what the level of asymmetry was in their femurs and tibiae; (2) which type of asymmetry predominates; and (3) whether fleas that fed on host species distantly related to their principal host species produced offspring that exhibited greater asymmetry compared with offspring of fleas that fed on their principal host species. We found fluctuating asymmetry in femurs and tibiae of X. ramesis and in the tibiae of P. chephrenis as well as significantly left-handed directional asymmetry in the femurs of P. chephrenis Host species identity significantly impacted asymmetry in leg segments of P. chephrenis but not in those of X. ramesis Offspring asymmetry increased when mother fleas fed on a host that was distantly related to the principal host. Fleas parasitizing multiple host species might compensate for developmental instability when utilizing a novel host species; therefore, host-switching events in host-specific parasites could be constrained by the relatedness between a novel and a principal host species.