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Differential diagnosis between Tunga penetrans (L., 1758) and T. trimamillata Pampiglione et al., 2002 (Insecta, Siphonaptera), the two species of the genus Tunga parasitic in man.

Of the ten currently known species of sand fleas, only two, Tunga penetrans and Tunga trimamillata, are known to be parasites of man, besides other warm blooded animals, most of which are peridomestic. The hosts of the other eight are limited to a few genera of wild mammals. T. trimamillata was only recently identified and differentiated from T. penetrans by features of the gravid female phase. In the present paper the different morphological characters of both for non-gravid females and males of the two species are described. In the non-gravid-females the distinguishing characters are: a) differences in the length of the first two segments of the maxillary palps (the first is the longest in T. trimamillata, whereas the second is the longest in T. penetrans), this feature is associated with another character i.e. the presence of short, thick spines in addition to the bristles on the surface of the segments only in T. trimamillata; b) the presence, only in T. trimamillata, of a row of spines on the antero-medial surface of the libia of the 3rd pair of legs; c) the last abdominal spiracle protrudes in T. trimamillata but not in T. penetrans; d) the hood of the hilla in spermatheca is surrounded by a papilla only in T. penetrans. The following morphological characters differentiate males of T. trimamillata and T. penetrans: a) T. trimamillata has a row of spines on the antero-medial surface of the tibia of the 3rd pair of legs; b) the diameter of the abdominal spiracles of T. trimamillata is smaller than that of T. penetrans and the edges of the spiracles are more regular; c) the claspers and aedeagus of T. trimamillata are shorter than those of T. penetrans; d) the profile of the oedeagal apodeme differs between the two species.

Animals↗

Molecular biological investigations of Brazilian Tunga sp. isolates from man, dogs, cats, pigs and rats.

Twenty-four isolates of Tunga were collected from afflicted humans, dogs, cats, pigs and rats in Brazil. To investigate genetic diversity, a hypervariable section of mitochondrial 16S rDNA was amplified using PCR and subsequently sequenced. In order to compare results with another species of the genus Tunga, three isolates of the recently described Tunga trimamillata were also checked. Whereas eleven isolates (five from cats, three from dogs and three from humans) were of identical sequence, thirteen isolates collected from dogs, humans, pigs and rats showed differences in sequence up to 49%, so that the existence of one or more new species of Tunga may be presumed.

Animals↗

Blood-feeding of Tunga penetrans males.

The jigger Tunga penetrans (Linnaeus, 1758: type-species of the family Tungidae) is the smallest known species of flea (Siphonaptera), causing serious ectoparasitosis of humans and domestic animals. The adult female Tunga lodges in the epidermis of the mammalian host, grows by neosomy, becomes gravid and expels eggs. Relatively little is known about the free-living male Tunga adults. Among impoverished communities of Fortaleza in north-east Brazil, we observed T. penetrans males as well as females penetrating the skin of human hosts. After penetrating the epidermis for a few hours, evidently for capillary feeding from the dermis, males withdrew their mouthparts and crawled away, whereas the females remained completely embedded, hypertrophying to become gravid, eventually dying in situ after oviposition. Caged rats were placed on the sandy soil and examined periodically for Tunga infestation. On five rats we obtained 140 females embedded and we detected 75 males biting, with rat erythrocytes observed in the proventriculus and midgut of all five males dissected and examined microscopically. This confirms that T. penetrans males are hamatophagous ectoparasites of mammals.

Animals↗

Additional description of a new species of Tunga (Siphonaptera) from Ecuador.

A new species of chigoe flea belonging to the genus Tunga Jarocki, 1838 (Siphonaptera, Pulicidae, Tunginae), Tunga trimamillata, was recently described by Pampiglione et al. (2002). A better description and more details of the epidemiology of the flea are now presented. It is a species found in goats, pigs and cattle in Santa Isabel in Andean Ecuador. This new species differs from its most similar congener, Tunga penetrans (L., 1758), by several features, the most important of which are a) the presence on the anterior extremity of the gravid female of three rounded humps surrounding the head and thorax (which, however, are not visible if viewed in profile), b) slightly larger dimensions and c) the length of the first segment of the maxillary palpi which is longer than each of the other three.

Animals↗

Wolbachia infection in the newly described Ecuadorian sand flea, Tunga trimamillata.

Wolbachia pipientis is an intracellular endosymbiont producing reproductive alterations in its hosts. This bacterium have been reported in many arthropods and nematodes. By PCR amplification and sequencing of the 16S rDNA and ftsZ genes we have identified a Wolbachia strain in the newly described sand-flea, Tunga trimamillata. Prevalence of this endosymbiont in the 26 individuals screened is equal to 35%. Sympatric and allopatric specimens of the related species Tunga penetrans were also analysed, but in contrast to literature data, Wolbachia appears absent in the presently analysed 24 specimens. Field studies evidence a female-biased sex-ratio in T. trimamillata, suggesting that Wolbachia may cause sex-ratio distortion in this species. By means of BLAST search and phylogenetic analysis we found that the Wolbachia strain from T. trimamillata pertains to the arthropod-infecting Wolbachia; this strain is highly differentiated from the Wolbachia strain of T. penetrans described in literature.

Animals↗

Tunga penetrans. A case report and review of the literature.

Tungiasis is an infection caused by infestation of Tunga Penetrans into the epidermis. Although, at the moment, presentation of this condition seems rare in Europe and Northern America, tungiasis is highly endemic in other parts of the world. We describe a case of Tunga Penetrans infection in the foot of a 22-year old Dutch female returning from Kenya, and present a review of the literature. Now that intercontinental travel is increasing, physicians in other parts of the world will be expected to recognize and treat this exotic infection. Knowledge of human tungiasis is mandatory in order to recognize the symptoms, achieve correct diagnosis and ensure adequate medical treatment to our patients.

Adult↗

Infestation by Tunga penetrans.

Infestation by the flea Tunga penetrans, although quite prevalent in Central America, South America, and Africa, is rarely recognized or reported in North America. This report describes a case of tungiasis occurring in a New York resident who acquired the disease in Africa. The patient manifested characteristic lesions of white papules with central black pits on the dorsa of her toes. Treatment consisted of removal of the offending parasites.

Adult↗

Investigations on the biology, epidemiology, pathology and control of Tunga penetrans in Brazil: I. Natural history of tungiasis in man.

Tungiasis is an important health problem in poor communities in Brazil and is associated with severe morbidity, particularly in children. The causative agent, the female flea Tunga penetrans, burrows into the skin of its host, where it develops, produces eggs and eventually dies. From the beginning of the penetration to the elimination of the carcass of the ectoparasite by skin repair mechanisms, the whole process takes 4-6 weeks. The present study is based on specimens from 86 patients, for some of whom the exact time of penetration was known. Lesions were photographed, described in detail and biopsied. Biopsies were examined histologically and by means of scanning electron microscopy (SEM). Based on clinical, SEM and histological findings, the "Fortaleza classification" was elaborated. This allows the natural history of tungiasis to be divided into five stages: (1) the penetration phase, (2) the phase of beginning hypertrophy, (3) the white halo phase, (4) the involution phase and (5) residues in the host's skin. Based on morphological and functional criteria, stages 3 and 4 are divided into further substages. The proposed Fortaleza classification can be used for clinical and epidemiological purposes. It allows a more precise diagnosis, enables the assessment of chemotherapeutic approaches and helps to evaluate control measures at the community level.

Aged↗

Investigations on the biology, epidemiology, pathology and control of Tunga penetrans in Brazil: III. Cytokine levels in peripheral blood of infected humans.

Tungiasis is caused by penetration of the female jigger flea, Tunga penetrans, into the skin of its host. This parasitic skin disease is almost invariably associated with intense inflammation around embedded fleas, the underlying mechanisms being unknown. A study was undertaken to determine whether the inflammatory process can be attributed to immune activation induced by a biologically active foreign body. We determined the concentrations of Th1-mediated (IFN-gamma, TNF-alpha) and Th2-mediated (IL-4) cytokines in the sera of patients with tungiasis. The results were compared with those of controls infected with different helminths or exposed to soil-transmitted helminths. The results show that tungiasis causes a mixed Th1 and Th2 immune response, characterized by significantly increased concentrations of the pro-inflammatory cytokines IFN-gamma and TNF-alpha, with a slightly increased concentration of IL-4. The preponderance of the Th1 immune response was indicated by a significantly increased TNF-alpha/IL-4 ratio in patients with tungiasis, as compared with the control groups.

Adolescent↗

Investigations on the biology, epidemiology, pathology and control of Tunga penetrans in Brazil: IV. Clinical and histopathology.

Tungiasis is a parasitic skin disease caused by the sand flea Tunga penetrans. This ectoparasitosis is endemic in South America, the Caribbean and sub-Saharan Africa, where it is an important but neglected health problem in resource-poor communities. As part of a study of tungiasis-related morbidity in a typical slum in Fortaleza, Brazil, we identified 86 individuals with tungiasis. Lesions were counted, classified according to the stage of development, and clinical pathology was documented. One hundred and nine lesions were biopsied and examined by histological sectioning. The patients had between 1 and 145 lesions (median 14.5), the majority occurring in clusters. In all, 77% of patients reported severe pain at the site of the lesion, and 52% had one or more nails lost or severely deformed. Intense inflammation and/or fissures hindered 45% of the patients from walking normally. Signs of superinfection were observed in 29%, and signs of generalized inflammation in 2% of patients. Clinical pathology was significantly related to the number of lesions, and the total number of parasites present correlated with the number of fleas occurring in clusters. Clinical pathology was frequently accompanied by a pathological alteration of the epidermis (predominantly hyperplasia, parakeratosis, hyperkeratosis, and spongiosis) and the dermis. Tungiasis causes a broad spectrum of clinical histopathological alterations, and is a serious health threat in a typical, impoverished community in northeast Brazil. The clinical pathology is closely related to the parasite burden of an individual and the clustering of embedded fleas at certain predilection sites.

Adolescent↗

Investigations on the biology, epidemiology, pathology and control of Tunga penetrans in Brazil. V. Cytokine concentrations in experimentally infected Wistar rats.

Tungiasis is caused by the penetration of the female sand flea Tunga penetrans into the skin of its host. This parasitic skin disease is almost invariably associated with an intense inflammation around embedded fleas, the underlying mechanisms being unknown. A study was undertaken to determine whether Wistar rats can be used as an animal model to assess cytokine kinetics during the natural course of the infection. Laboratory-raised Wistar rats were exposed in cages put on the soil in an area with high human attack rates. Rats were examined daily and blood samples were taken before exposure and at 2, 6, 10, 13, 16 and 20 days after flea penetration. TNF-alpha, IL-1 beta, IFN-gamma, IL-4, IL-10 and CINC (a rat cytokine- induced neutrophil chemoattractant and member of the IL-8 family) were determined by enzyme immunoassay. The results showed an increasing serum concentration of TNF-alpha and IL-1 beta 10-13 days after penetration and a rapid increase in IL-4 2 days after fleas became embedded. During the natural course of the infection, the ratio of the serum concentration of TNF-alpha to that of IL-10 decreased, indicating a relative increase in the secretion of the anti-inflammatory cytokine. The treatment of lesions with silicone oil abrogated the natural disease course and changed the pattern of cytokine secretion. We conclude that the Wistar rat is an appropriate model to study immune responses in tungiasis.

Animals↗

Field trial of the efficacy of a combination of imidacloprid and permethrin against Tunga penetrans (sand flea, jigger flea) in dogs in Brazil.

In a field trial in Brazil 17 dogs penetrated by females of the jigger flea, Tunga penetrans, were topically treated with a combination of 10% imidacloprid and 50% permethrin (Advantix), while 17 dogs remained untreated. The follow-up controls on days 7, 14, 21 and 28 post-treatment clearly showed that, beginning from day 7, the flea load in treated dogs decreased, so that most of the dogs became free of tungiasis lesions, while in the untreated group the flea load remained high. Since the dogs distribute the flea eggs throughout the village, leading to a high incidence of tungiasis in humans, treatment of dogs probably also decreases the number of cases of tungiasis in the latter.

Animals↗

Genetic variability of Tunga penetrans (Siphonaptera, Tungidae) sand fleas across South America and Africa.

Tunga penetrans is a widely distributed sand flea, infecting men and domestic animals. It originated in South America, but it is now also endemic of Sub-Saharan Africa due to a recent accidental introduction. Previous genetic analyses indicated a very limited variability in the Ecuadorian populations; on the other hand, samples from Madagascar resulted to be genetically isolated. To better examine the genetic variability of T. penetrans, a wider sampling was analysed for mitochondrial (cytochrome oxidase II) and nuclear (ribosomal internal transcribed spacer [ITS] 2) DNA sequences. The mitochondrial marker confirms the low genetic variability, with few haplotypes characterizing the majority of individuals from different populations. Haplotype distribution is in agreement with a recent colonization of Africa and with a rapid spreading across this continent. Moreover, a consistent gene flow between the Pacific and Atlantic South American populations emerges, possibly due to human and/or animal cross-Andean dispersal. On the other hand, the ITS2 marker depicts a sharply diverging pattern with samples collected in Ecuador unequivocally distinguished from the Brazilian and African ones. On the whole, taking into account also the molecular features of the marker used, data here presented are better interpreted in the light of a high dispersal ability of T. penetrans, probably reducing the phylogeographic signal.

Africa↗

High infection rate of Wolbachia endobacteria in the sand flea Tunga penetrans from Brazil.

Tunga penetrans is an ectoparasite causing considerable morbidity in endemic communities. Recently, endobacteria of the genus Wolbachia were identified also in T. penetrans. Since Wolbachia were suggested as targets for intervention of insect pests and human filariasis, sand fleas were collected from infested humans, dogs and rats in a hyperendemic area in northeastern Brazil, and screened for Wolbachia infections. Twenty-one adult fleas and four batches of flea eggs were examined by PCR using primers targeting the 16S rDNA, the DNA coding for FtsZ cell-cycle protein or a Wolbachia surface protein (WSP-1). Wolbachia were detected in all examined samples from eggs, free-living male and female fleas and from neosomic female fleas. No Wolbachia DNA was detected in two samples containing flea faeces. In addition, Wolbachia were labelled by immunohistology in the ovaries of 37 female fleas using antisera raised against WSP-1 of Wolbachia the filarial parasite Dirofilaria immitis. In the vicinity of the embedded fleas containing the Wolbachia, infiltrations of neutrophils and macrophages were observed. This study showed that Wolbachia endobacteria are abundant in T. penetrans and that all examined fleas were infected by these endobacteria. Our findings may have important implications for the future development of control strategies for human tungiasis.

Animals↗

A plant-based repellent protects against Tunga penetrans infestation and sand flea disease.

Tungiasis is a parasitic skin disease prevalent in impoverished populations in the tropics and associated with considerable morbidity. Treatment options are limited and prophylaxis has never been attempted. We assessed the effectiveness of a plant-based repellent to prevent infestation with Tunga penetrans and sand flea disease in an urban squatter settlement in Fortaleza, Northeast Brazil. Two cohorts were formed. One cohort started with the intervention while another served as control. The plant-based repellent Zanzarin was applied twice daily on the skin of the feet. After 4 weeks and a wash-out-phase of another 2 weeks, cross-over was performed. Study members were examined twice per week for the presence of newly embedded sand fleas. The number of viable and dead sand fleas and the total number of sand flea lesions were determined. Sentinel rats were used to assess local transmission dynamics. Outcome measures were the number of newly embedded sand fleas per individual per day, number of sand flea lesions per individual according to the stage, as well as type and degree of clinical pathology. Zanzarin applied twice daily on the feet reduced the number of newly embedded fleas by 92% (interquartile range 81-100%), the total number of lesions by 87% (81-98%) and reversed tungiasis-associated clinical pathology almost completely. In conclusion, in a setting in which tungiasis is an important health threat, the daily application of a plant-based repellent prevented the infestation with T. penetrans and sand flea disease.

Administration, Topical↗

Tunga penetrans: molecular identification of Wolbachia endobacteria and their recognition by antibodies against proteins of endobacteria from filarial parasites.

In search of Wolbachia in human parasites, Wolbachia were identified in the sand flea Tunga penetrans. PCR and DNA sequencing of the bacterial 16S rDNA, the ftsZ cell division protein, the Wolbachia surface protein (wsp) and the Wolbachia aspartate aminotransferase genes revealed a high similarity to the respective sequences of endosymbionts of filarial nematodes. Using these sequences a phylogenetic tree was generated, that indicates a close relationship between Wolbachia from T. penetrans and from filarial parasites, but possibly as a member of a new supergroup. Ultrastructural studies showed that Wolbachia are abundant in the ovaries of neosomic fleas, whereas other, smaller and morphologically distinct, bacteria were observed in the lumen of the intestine. Wolbachia were labeled by immunohistology and immunogold electron microscopy using polyclonal antibodies against wsp of Drosophila, of the filarial parasite Dirofilaria immitis, or against hsp 60 from Yersinia enterocolitica. These results show that as in filariasis, humans with tungiasis are exposed to Wolbachia. Furthermore, antisera raised against proteins of Wolbachia from arthropods or from filarial parasites can be immunologically cross-reactive.

Amino Acid Sequence↗