Prevalence of Sarcoptes scabiei in Patients with Suspected scabies

Objective: Scabies is caused by an ectoparasite called Sarcoptes scabiei (S. scabiei), which penetrates the epidermis through skin folds and burrows in the stratum corneum, following the development of tunnels (sillion). The disease is specifically characterised by keratosis, allergy and itching that increases at night-time. This study aimed to investigate the frequency of S. scabiei in patients with a pro-diagnosis of scabies. Objective: Between January 2012 and December 2019, a total of 746 [n=388 (52%), female; n=358 (48%) male] patients aged 0-80 years were admitted to Firat University Hospital Parasitology-mycology Laboratory. Skin scrapings were taken from suspected lesions on anatomic regions such as the hands (wrist, interdigital skin, fingertip and palm), abdomen, penis and legs (thigh and bottom foot). They were examined under a light microscope after adding 15% potassium hydroxide solution. Results: S. scabiei was positive in 139 (18.63%) of 746 patients including a mother and her daughter and a married couple, where 68 (9.11%) were female and 71 (9.52%) were male. Conclusion: To our best knowledge, this is the first comprehensive study of scabies in Elazig. Despite the recent socio-economic and cultural developments observed in our country, scabies and all other parasitic infestations still remain to be important problems. We believe that improvement of the public vigilance together with early diagnosis will improve sanitation and provide protection against scabies and parasitic infestations.

Costello syndrome model mice with a HrasG12S/+ mutation are susceptible to develop house dust mite-induced atopic dermatitis.

Costello syndrome is an autosomal dominant disorder that is caused by germline HRAS mutations. Patients with Costello syndrome present craniofacial abnormalities, cardiac defects, and cancer predisposition, as well as skin abnormalities, including papillomas, keratosis pilaris, and eczematous dermatitis. However, the mechanisms underlying the dermatological abnormalities remain unclear. Here, we demonstrated that knock-in mice expressing an Hras G12S mutation (HrasG12S/+ mice) are susceptible to develop atopic dermatitis (AD)-like skin lesions, including eczema, pruritus, elevated serum IgE levels, acanthosis, and the infiltration of mast cells, basophils, and type-2 innate lymphoid cells in the dermis, after stimulation with house dust mite allergens (Dermatophagoides farinae, Dfb). Reduced skin barrier function, increased proliferation of phosphorylated ERK (p-ERK)-positive epidermal cells, and increased Th2-type cytokines as well as epithelial cell-derived cytokines, including IL-33, were observed in the skin tissue of HrasG12S/+ mice compared with Hras+/+ mice. Cultured HrasG12S/+ keratinocytes exhibited increased IL-33 expression after Dfb stimulation. PD0325901, an MEK inhibitor, ameliorated AD-like symptoms in HrasG12S/+ mice, showing decreased proliferation of p-ERK-positive epidermal cells and decreased expression of IL-33. Our findings indicate that the epidermis of HrasG12S/+ mice stimulated by Dfb strongly induced IL-33 expression and type-2 innate lymphoid cells, resulting in AD-like skin lesions. These results suggest that the epidermis of HrasG12S/+ mice are prone to development of eczematous dermatitis stimulated with house dust mite allergens.

Transcriptional Analysis of Human Skin Lesions Identifies Tryptophan-2,3-Deoxygenase as a Restriction Factor for Cutaneous Leishmania.

Disease manifestation after infection with cutaneous Leishmania species is the result of a complex interplay of diverse factors, including the immune status of the host, the infecting parasite species, or the parasite load at the lesion site. Understanding how these factors impact on the pathology of cutaneous leishmaniasis (CL) may provide new targets to manage the infection and improve clinical outcome. We quantified the relative expression of 170 genes involved in a diverse range of biological processes, in the skin biopsies from patients afflicted with CL caused by infection with either L. major or L. tropica. As compared to healthy skin, CL lesions bear elevated levels of transcripts involved in the immune response, and conversely, present a significant downregulation in the expression of genes involved in epidermal integrity and arginine or fatty acid metabolism. The expression of transcripts encoding for cytotoxic mediators and chemokines in lesions was inversely correlated with the expression of genes involved in epidermal integrity, suggesting that cytotoxicity is a major mediator of CL pathology. When comparing the transcriptional profiles of lesions caused by either L. major or L. tropica, we found them to be very similar, the later presenting an aggravated inflammatory/cytotoxic profile. Finally, we identified genes positively correlated with the parasite load in lesions. Among others, these included Th2 or regulatory cytokines, such as IL4 or IL10. Remarkably, a single gene among our dataset, encoding for tryptophan-2,3-deoxygenase (TDO), presented a negative correlation with the parasite load, suggesting that its expression may restrict parasite numbers in lesions. In agreement, treatment of macrophages infected with L. major in vitro with a TDO inhibitor led to an increase in parasite transcripts. Our work provides new insights into the factors that impact CL pathology and identifies TDO as a restriction factor for cutaneous Leishmania.

[Henneguya wolinensis (Myxosporea: Myxobolidae), a new for Russian fauna parasite from the perch Perca fluviatilis L.].

The infection of the perch Perea fluviatilis L. with myxosporean Henneguya wolinensis Romuk-Wodoracki, 1990 has been detected. This is the second finding of this parasite after its original descriptin and the first for Russia. Plasmodium of this species develops in the epidermis under scales throughout the body causing the formation of white cysts up to 1 mm. Spores are fusiform, large, their average length constitutes 25.5 µm without the caudal appendages and 62 µm with them. Slight morphological differences in spore structure comparing to original description have been revealed.

Cutaneous histoplasmosis with prominent parasitization of epidermal keratinocytes: report of a case.

Disseminated histoplasmosis most commonly occurs in immunosuppressed individuals and involves the skin in approximately 6% of patients. Cutaneous histoplasmosis with an intraepithelial-predominant distribution has not been described. A 47-year-old man was admitted to our institution with fever and vancomycin-resistant enterococcal bacteremia. He had been diagnosed with T-cell prolymphocytic leukemia 4 years earlier and had undergone matched-unrelated-donor stem cell transplant 2 years earlier; on admission, he had relapsed disease. His medical history was significant for disseminated histoplasmosis 6 months before admission, controlled with multiple antifungal regimens. During this final hospitalization, the patient developed multiple 2-5 mm erythematous papules, a hemorrhagic crust across the chest, shoulders, forearms, dorsal aspect of the fingers, abdomen and thighs. Skin biopsy revealed clusters of oval yeast forms mostly confined to the cytoplasm of keratinocytes and within the stratum corneum; scattered organisms were present in the underlying superficial dermis without any significant associated inflammatory infiltrate. Special stains and immunohistochemical studies confirmed these to be Histoplasma organisms. We highlight this previously unrecognized pattern of cutaneous histoplasmosis to ensure its prompt recognition and appropriate antifungal therapy.

Teleost skin, an ancient mucosal surface that elicits gut-like immune responses.

Skin homeostasis is critical to preserve animal integrity. Although the skin of most vertebrates is known to contain a skin-associated lymphoid tissue (SALT), very little is known about skin B-cell responses as well as their evolutionary origins. Teleost fish represent the most ancient bony vertebrates containing a SALT. Due to its lack of keratinization, teleost skin possesses living epithelial cells in direct contact with the water medium. Interestingly, teleost SALT structurally resembles that of the gut-associated lymphoid tissue, and it possesses a diverse microbiota. Thus, we hypothesized that, because teleost SALT and gut-associated lymphoid tissue have probably been subjected to similar evolutionary selective forces, their B-cell responses would be analogous. Confirming this hypothesis, we show that IgT, a teleost immunoglobulin specialized in gut immunity, plays the prevailing role in skin mucosal immunity. We found that IgT(+) B cells represent the major B-cell subset in the skin epidermis and that IgT is mainly present in polymeric form in the skin mucus. Critically, we found that the majority of the skin microbiota are coated with IgT. Moreover, IgT responses against a skin parasite were mainly limited to the skin whereas IgM responses were almost exclusively detected in the serum. Strikingly, we found that the teleost skin mucosa showed key features of mammalian mucosal surfaces exhibiting a mucosa-associated lymphoid tissue. Thus, from an evolutionary viewpoint, our findings suggest that, regardless of their phylogenetic origin and tissue localization, the chief immunoglobulins of all mucosa-associated lymphoid tissue operate under the guidance of primordially conserved principles.

The nipple-areola complex epidermis: a prospective systematic study in adult autopsies.

The prevalence of different types of clear cells and of the mite Demodex in the nipple-areola complex of adult autopsies of both sexes not suffering from breast cancer was studied in a total of 140 nipples. The epidermis of the nipple-areola complex shows squamous cells and 3 types of clear cells: Toker cells, pagetoid dyskeratosis cells, and signet ring-like cells. Toker cells were identified by standard light microscopy in 13 of 140 nipples (9.3%). Reactivity of these cells for CK7 was observed in 35 nipples (25%). They are derived from the lactiferous duct epithelium. Pagetoid dyskeratosis cells were identified in 56 of 140 nipples (40%). In 12 nipples, these cells were conspicuous (8.6%). It is suggested that the proliferation of these cells is induced by friction. Signet ring-like cells were identified in 71 nipples (50.7%). In 2 nipples, these cells were conspicuous (1.4%). They are a consequence of artefact related to formalin fixation. The prevalence of all these clear cells has no relationship with gender. Routine histopathological examination is usually enough to distinguish the characteristic features of the clear cells involving the nipple epidermis and permits differentiation of other entities with epidermal pale cells. Demodex mites were observed in 58 nipple-areola complexes (41.4%). They were more common in male nipple-areola complexes (P < 0.05). The prevalence of these mites was seen to remain steady along the years since the third decade. Demodex mites are common parasites of human nipple and are apparently of no pathologic significance.

Paranucleospora theridion n. gen., n. sp. (Microsporidia, Enterocytozoonidae) with a Life Cycle in the Salmon Louse (Lepeophtheirus salmonis, Copepoda) and Atlantic Salmon (Salmo salar).

Paranucleospora theridion n. gen, n. sp., infecting both Atlantic salmon (Salmo salar) and its copepod parasite Lepeophtheirus salmonis is described. The microsporidian exhibits nuclei in diplokaryotic arrangement during all known life-cycle stages in salmon, but only in the merogonal stages and early sporogonal stage in salmon lice. All developmental stages of P. theridion are in direct contact with the host cell cytoplasm or nucleoplasm. In salmon, two developmental cycles were observed, producing spores in the cytoplasm of phagocytes or epidermal cells (Cycle-I) and in the nuclei of epidermal cells (Cycle-II), respectively. Cycle-I spores are small and thin walled with a short polar tube, and are believed to be autoinfective. The larger oval intranuclear Cycle-II spores have a thick endospore and a longer polar tube, and are probably responsible for transmission from salmon to L. salmonis. Parasite development in the salmon louse occurs in several different cell types that may be extremely hypertrophied due to P. theridion proliferation. Diplokaryotic merogony precedes monokaryotic sporogony. The rounded spores produced are comparable to the intranuclear spores in the salmon in most aspects, and likely transmit the infection to salmon. Phylogenetic analysis of P. theridion partial rDNA sequences place the parasite in a position between Nucleospora salmonis and Enterocytozoon bieneusi. Based on characteristics of the morphology, unique development involving a vertebrate fish as well as a crustacean ectoparasite host, and the results of the phylogenetic analyses it is suggested that P. theridion should be given status as a new species in a new genus.

Subcutaneous palisading granulomatous pseudocysts of Echinococcus granulosus origin.

BACKGROUND: Palisading granulomatous reactions are documented in many diseases. Although subcutaneous cystic echinococcosis (CE) is documented rarely, a subcutaneous palisading, granulomatous, pseudocystic (PGP) reaction to elusive Echinococcus granulosus membranous components, in the absence of cutaneous fistulization, is undocumented. METHODS: Seven-year clinicopathological review of subcutaneous echinococcal PGP reactions. RESULTS: Gross: seven thick-walled 'cysts' containing clear or straw-colored fluid were investigated. Histopathology: the pseudocysts contained a palisade of epithelioid histiocytes and giant cells. Focal periodic acid Schiff (PAS)-positive eosinophilic fragments, some resembling keratin 'flakes', were identified within the lumen or within the cellular palisade, consistent with a PGP reaction to fragmented E. granulosus membrane. Clinical correlation: the initial histopathological diagnosis of two patients was ruptured epidermoid cysts with an assumed granulomatous reaction to eosinophilic keratinous debris. Subsequent diagnosis of CE in the liver and cervical soft tissue necessitated review of the 'epidermoid cysts'; PAS-positive E. granulosus membranous fragments masqueraded as keratinous debris. Renal, hepatic and pleuropulmonary CE were confirmed in the remaining patients following confirmation of an echinococcal PGP reaction. CONCLUSION: Heightened awareness and obsessive appraisal of subcutaneous PGP reactions for subtle, focal, PAS-positive and echinococcal membranous fragments are pivotal to the diagnosis that also serves as a clue to visceral CE.

Epidermal 'alarm substance' cells of fishes maintained by non-alarm functions: possible defence against pathogens, parasites and UVB radiation.

Many fishes possess specialized epidermal cells that are ruptured by the teeth of predators, thus reliably indicating the presence of an actively foraging predator. Understanding the evolution of these cells has intrigued evolutionary ecologists because the release of these alarm chemicals is not voluntary. Here, we show that predation pressure does not influence alarm cell production in fishes. Alarm cell production is stimulated by exposure to skin-penetrating pathogens (water moulds: Saprolegnia ferax and Saprolegnia parasitica), skin-penetrating parasites (larval trematodes: Teleorchis sp. and Uvulifer sp.) and correlated with exposure to UV radiation. Suppression of the immune system with environmentally relevant levels of Cd inhibits alarm cell production of fishes challenged with Saprolegnia. These data are the first evidence that alarm substance cells have an immune function against ubiquitous environmental challenges to epidermal integrity. Our results indicate that these specialized cells arose and are maintained by natural selection owing to selfish benefits unrelated to predator-prey interactions. Cell contents released when these cells are damaged in predator attacks have secondarily acquired an ecological role as alarm cues because selection favours receivers to detect and respond adaptively to public information about predation.

Amebiasis cutis revisited.

BACKGROUND: Amebiasis cutis (AC) is reported infrequently. This study assesses the clinicopathological spectrum, co-existent visceral involvement and impact of human immunodeficiency virus (HIV) co-infection on AC. METHODS: An 8-year prospective clinicopathological evaluation of patients with AC. RESULTS: Thirty-one biopsies of ulcers, fistulae, fissures, abscesses, polypoid and warty lesions in perianal, penile, scrotal, vulval, buttock, chest and abdominal wall sites were evaluated. Of these, 11 had a 'superficial' (superficial AC) and 20 a 'deep' (deep AC), histopathological pattern. Superficial AC showed predominant epidermal spongiosis, liquefactive necrosis, ulceration and fissures with hematophagous amebic trophozoites (HATs). Deep AC had confluent deep dermal and subcutaneous liquefactive, coagulative or suppurative necrosis and HATs. Seven biopsies showed vasculitis or thrombosis with luminal HATs. OUTCOME: Fourteen patients died; 9 had concomitant visceral amebiasis, 5 had other co-infections. Six who died were HIV seropositive, three were seronegative; all had deep AC. Of the 17 survivors, 11 (8 HIV positive) had superficial AC that healed with metronidazole treatment; the remaining 6 (one HIV seropositive) required additional surgical intervention. CONCLUSION: Deep AC is predictive of co-existent, contiguous visceral disease. The effective management, histopathological mimickers and diagnostic pitfalls of superficial and deep AC differ. The outcome in HIV-infected patients is dependent on co-existent systemic diseases.

Cutaneous leishmaniasis: evaluation by polymerase chain reaction in the Cukurova region of Turkey.

The diagnosis of cutaneous leishmaniasis (CL) may depend on the detection of the parasite in histologic sections, the growth of the promastigotes in culture, or the identification of parasite by other techniques. We performed polymerase chain reaction (PCR) on paraffin-embedded biopsies to determine the validity of this technique for diagnosis of CL. PCR was used to detect the parasite using 2 different DNA extraction methods. PCR was positive in all 20 cases when the Leishmania parasite was detected by light microscopy. Twenty-seven of 34 cases that were negative microscopically for the parasite were positive using PCR. The first extraction method of DNA identified leishmanial DNA in 41 of 54 cases (75.9%); the second extraction of DNA was positive in 47 of 54 cases (87%). PCR was negative in all of the nonleishmaniasis cases. The PCR-based method appears to be a useful diagnostic approach for identification of suspected cases of CL.

Frequency of epidermal langerhans cells in site of intradermal injection promastigotes of leishmania braziliensis in mice

We utilized inmunostaining assay with avidein boitin peroxidase technique and primary rat monoclonal antibody NLDC-145 specific for mouse dentritic cell, characterized langerhans cells (LC) in epidermis sheet of 1 mm² of skin of the footpads of inbred females BALB/c mice, intradermally (id) injected with 1.10ü cultured promastigotes of leishmania braziliensis. The result showed that the injection of the parasites in the skin of the animals produced a progressive increment of epidermal LC in the site of the previus injection, and statistically significant (P<0.05) in each study period. The density of epidermal LC was going up by parasite insult in early time, after the 15 minutes (m) post-infection (pi) with 162ñ31.2 LC/mm², reaching a maximal value of 4503ñ713 LC/mm² to 2 week (w) pi until 898ñ481 LC/mm² to 6 w pi. The number of LC was always higher in epidermis of the control healthy mice, these skin samples showed 120ñ28.9 LC/mm². Morphological changes of the promastigotes injected could to be detected in skin giemsa-stained imprints, the parasite showed form ovoid and a short flagel, between 15m and 2 hour (hr) pi. No parasites were even seen in the imprint samples than of as: activated macrfophages (33 percent) to 15 m pi, neutrophils (46,33 percent) to 4 hr pi, eosinophils (2,33 percent) to 4 hr pi, lymphocytes (15,67 percent) to 6 hr pi, degraded lymphocytes (8,33 percent) to 1 w pi, monocytes (4,33 percent) to 4 day (d) pi and activated monocytes (19,33 percent) to 1 d pi. The stained sections of skin inoculated, revealed amastigotes into macrophages dermal near of the perivascular area and inflammatory process in the dermis consisted of lymphocytes, monocytes, plasma cell and polymorphonuclear cells between 1 w and 6 w pi. No parasites were detected in the epidermis. the results showed that the promastigotes in the skin survived the first 2 hr out of the macrophages, and on the other hand, stinuled various cell types in sites of injection of the parasites, and the proliferation of antigen presentation by epidermalk langerhans cells, necessary for the initiation of the specific T cell inmune response

Shell disease in river cooters (Pseudemys concinna) and yellow-bellied turtles (Trachemys scripta) in a Georgia (USA) lake.

A disfiguring shell disease was detected in river cooters (Pseudemys concinna) and yellow-bellied turtles (Trachemys scripta) from Lake Blackshear, Georgia (USA). The turtles used were part of a mark-recapture study conducted from September 1991 to June 1993. Histologic changes on four turtles included acute segmental necrosis of the epidermis, followed by ulceration, necrosis of the underlying dermis and dermal bone, and exaggerated remodeling of bone. Additional findings included visceral inflammatory lesions and bacterial infection, sepsis and marked trematode ova granulomatosis. The cause of the shell lesions was not determined.

In situ characterization of the human host response to Leishmania panamensis.

Both host and parasite determinants influence the outcome of Leishmania infections. Human host responses in cutaneous leishmaniasis of limited duration caused by a single species of the Viannia (V) subgenus were studied in skin biopsies obtained from lesions caused by Leishmania (V) panamensis in 31 male patients from the Colombian Pacific Coast. Dermal infiltrates and histopathologic changes were characterized using monoclonal antibodies and an indirect immunoperoxidase method. Dermal distribution of T-cell subpopulations and B-lymphocytes was nonrandom: CD4+ and CD8+ T cells were most frequent in the upper dermis, and B cells were most abundant in the lower dermis. Parasites, macrophages, and neutrophils were localized predominantly in the middermis. Multiple regression analyses to establish associations between lesion type (ulcer, nodule, or papule), immune response data (Montenegro skin test, indirect fluorescence antibody test titers, lymphocyte blastogenesis), and particular cell populations demonstrated statistically significant correlations between CD4+ lymphocytes and macrophages (p < 0.05). CD8+ lymphocytes were associated with plasma cells (p < 0.001), as was the presence of amastigotes (p < 0.05). These associations and the in situ divergence of CD4 and CD8 ratios suggest that prognostic indicators for disease evolution could be identified by prospective analysis of cellular relationships and response to therapy.

Leishmaniasis presenting as a dermatomyositis-like eruption in AIDS.

Three patients are described with leishmaniasis and AIDS, with cutaneous lesions mimicking dermatomyositis. Leishmania organisms were observed in great numbers in the dermis of lesional skin biopsy specimens. They were also present inside keratinocytes in all layers of the epidermis in one patient. Skin cultures from all patients and bone marrow culture in patients 1 and 3 revealed Leishmania infantum. Leishmania organisms were also found in nonlesional skin. The absence of proximal symmetric muscle weakness, elevated muscle enzymes, myopathic electromyograms, or characteristic histopathologic and immunologic features of dermatomyositis, and the rapid and complete clearance or marked improvement of the cutaneous lesions after treatment for leishmaniasis, make us consider true dermatomyositis unlikely. We suggest that leishmaniasis be included in the list of diseases capable of inducing a dermatomyositis-like eruption.

[Histopathology of mucocutaneous leishmaniasis caused by Leishmania (Vianna) braziliensis]. / Histopathologie de la leishmaniose cutanéo-muqueuse à Leishmania (Viannia) braziliensis.

A histopathological study of mucocutaneous leishmaniasis due to Leishmania (Viannia) braziliensis was carried out on 28 cutaneous and 114 mucosal biopsies, taken from Bolivian and Peruvian patients. This study showed similar histopathological findings in cutaneous and mucosal lesions. The cutaneous biopsies showed a strong epidermal hyperplasia occasionnally budding in the dermis. In the ulcerative area, the epidermis was totally necrosed and replaced by a fibrino-leucocytic edge. In the dermis, histio-lympho-plasmocytic infiltration was constantly found. The histiocytes often gathered in follicles sometimes with diffuse fibrosis. The parasites were encountered in 28.6 p. 100 of the biopsies. Whatever the mucosa concerned (i.e. nasal, palatal or lingual), the mucosal lesion was not different from the cutaneous lesion. The malpighian epithelium is either absent or the seat of a pseudo-epitheliomatous hyperplasia. Major histio-lympho-plasmocytic infiltration was found and extended through the depth of the lamina propria. Suppurative and fibrinoid necroses coexisted superficially and sometimes penetrated in depth. The parasites were found in about 30 p. 100 of the cases.

Microfilaricidal effect of amocarzine in skin punch biopsies of patients with onchocerciasis from Latin America.

Skin punch biopsies were performed in 54 selected patients with onchocerciasis participating in a clinical trial with amocarzine (CGP 6140) in Ecuador and Guatemala. Skin snipping for counting microfilariae of Onchocerca volvulus was done before treatment (day 0) and day 4 and 8 following start of the therapy which consisted of 3 mg/kg amocarzine postprandially twice daily for three consecutive days. The mean microfilarial skin density has been reduced by 45% on day 4 and 95% on day 8. Skin punch biopsies were taken on day 5, within 1 cm from the snip site on the iliac crest. Histopathologic examination revealed that the vast majority of the microfilariae in the upper as well as in the deeper dermis were degenerated or necrotic, surrounded often (57%) by minute foci of fibrinoid change of the collagen. There was usually slight, less frequently moderate eosinophilic, lympho-plasmocytic and initial histocytic inflammatory reaction in the vicinity. Microfilariae were frequently (69%) found at the dermal-epidermal junction and in the epidermis. Occasionally (7%) intra-epidermal microabscesses were noted. Microfilariae were detected also in the lumen of some dermal lymphatic vessels. Therefore it is concluded that amocarzine showed marked microfilaricidal effects in the skin of patients with onchocerciasis as evidenced histologically by mainly destroyed or moribund microfilariae which induced a mild to moderate inflammatory cell reaction.

Leishmania (Viannia) braziliensis: comparative pathology of golden hamsters infected with isolates from cutaneous and mucosal lesions of patients residing in Tres Bracos, Bahia, Brazil.

The histopathology of primary forepaw and metastatic lymph node, spleen, and liver lesions produced in golden hamsters infected with cutaneous leishmaniasis (CL) strains (LTB 111 and LTB558) and mucocutaneous leishmaniasis (MCL) strains (LTB12 and LTB201) of Leishmania (Viannia) braziliensis isolated from patients residing in Tres Bracos, Bahia, Brazil is described. No pathological features providing clear differentiation of the CL and MCL strains were found. Although amastigotes were plentiful early in the development of primary forepaw lesions, they were either absent or could not be identified with certainty in sections of late stage lesions. Similarly, amastigotes were not found in histologic lesions at metastatic sites; however, leishmanial DNA was detected in both early and late stage forepaw lesions and metastatic lesions using Leishmania kinetoplast DNA and the gene coding for gp63 as hybridization probes. The DNA recovered from metastatic lesions was extracted from formalin-fixed paraffin-embedded tissues that had been stored at room temperature for prolonged periods.

The epidermal location and possible feeding site of Psorergates ovis, the sheep itch mite.

Skin biopsies from two Merino sheep heavily infested with Psorergates ovis were immersed in liquid nitrogen and cut into vertical frozen sections stained with lipophilic Sudan IV or Oil Red O and haematoxylin. A survey of the lateral distribution of mite sections showed a majority (ca 80%) were in or within 0.2 mm of the follicle mouth. A survey of vertical distribution showed no mite penetration deeper than inner stratum corneum where 57% of mite sections were seen; 30% were within outer stratum corneum or scurf; 13% were on the outer surface and less than 1% were detached. Lipid was the only material seen within stained mites at a location considered to be gut. This was supported by dosing sheep with quinacrine, taking biopsies at Day 6 and Day 14 and examining frozen sections under blue light. Fluorescent lipid was seen at a location considered to be mite gut. From these results it was recommended that acaricide treatments against P. ovis be lipophilic and administered transepidermally because less than 15% of mites were at a superficial location likely to be reached by topical application.