Isolation, identification, and phylogenetic analysis of potentially pathogenic free-living amoebae isolated from nasal and oral mucosa of HIV/AIDS patients in Iran.

The burden of HIV/AIDS in Iran is not as high as in the other countries with high prevalence; however, the number of cases of HIV/AIDs is increasing in this region. According to a recent report, Iran had 5000 (1400-13,000) new cases of HIV infection with 4000 (2500-6200) AIDS-related deaths. Individuals affected by HIV/AIDS are highly susceptible for developing opportunistic infections, e.g. the cerebral complications related to pathogenic free-living amoebae and colonization of free-living amoebae (FLA) can be a serious hazard for patients living with HIV/AIDS. In the present study, a total of 70 oral and nasal mucosal samples were obtained from HIV/AIDS patients referred to the reference hospitals in Iran and tested for the presence of potentially pathogenic FLA using culture and PCR/sequencing-based methods. To discern the taxonomic status of Acanthamoeba genotypes a maximum likelihood phylogenetic tree was constructed and tolerance assays were performed for the positive Acanthamoeba strains. Among the patients with HIV/AIDS referred to the reference hospitals from 2017 to 2019, 7.1% were found positive for pathogenic free-living amoebae. Three strains (HA3, HA4, and HA5) belonged to the T4 genotype, one strain (HA1) was related to the T5 genotype assigned as A. lenticulata, and another strain (HA2) had high homology to Vermamoeba vermiformis. The tolerance assay used for Acanthamoeba strains (HA1, HA3, and HA4) classified these amoebae as highly pathogenic strains. For the most part, the encephalitis cases occurring in HIV/AIDS patients in Iran remain undiagnosed due to lack of awareness of the practitioners on the available diagnostic tools for this lethal infection; therefore, the true incidence of GAE in this region is unknown. A possible colonization with FLA should be considered in the differential diagnosis of suspected cases of CNS infections among HIV/AIDS patients. This study is the first worldwide comprehensive study attempting to isolate and identify the FLA colonization in HIV/AIDS patients. This study highlights the fact that clinicians should be aware of the differential diagnosis of cerebral disease related to FLA in patients with HIV/AIDS.

Cutaneous leishmaniasis by Leishmania infantum: behind granulomatous lesions of unknown aetiology.

BACKGROUND: Cutaneous leishmaniasis (CL) is underestimated in Spain as in other European countries due to the polymorphism of its clinical manifestations and histopathological features discouraging doctors from suspecting leishmaniasis. Mucosal manifestations (ML) are misdiagnosed due to the fact that they often mimic cancer. OBJECTIVES: Given that leishmaniasis may be masked as different granulomatous diseases in Leishmania infantum endemic areas, the aim of this study was to verify this misdiagnosing and contributes to the improvement of CL/ML diagnosis. METHODS: A retrospective study involving formalin-fixed paraffin-embedded tissue biopsies with histopathological features of granulomatous lesions of unknown origin (GLUO) detected in 17 patients. This study included 13 patients with CL that was used as positive controls, nine patients with other confirmed diseases used as negative controls and seven patients with histological features suggestive of CL or ML without confirmation. Molecular analysis was blindly performed using two different PCR techniques. RESULTS: The PCR detected 15 CL cases in which the diagnosis was neither clinically nor histologically suspected. Leishmaniasis was confirmed in seven suspected patients in whom the classical techniques failed to detect the parasite. L. infantum was identified in all cases. A systematic review of CL cases in GLUO patients from European countries identified 45 reported cases. CONCLUSIONS: In L. infantum endemic areas, a high percentage of GLUO are due to Leishmania infection. The main consequences are delayed diagnosis and underestimation of the real incidence. PCR performed on paraffin-embedded tissue proved to be a reliable tool for diagnosis of CL/ML and must be performed routinely in any granulomatous dermatitis, even when the morphological features are no stereotypical of leishmaniasis.

Three new species of Clinostomum Leidy, 1856 (Trematoda) from Middle American fish-eating birds.

We recently engaged in a two-part study of Clinostomum Leidy, 1856 across a geographic range comprising central Mexico southwards to Costa Rica, in Central America. In the first study, we investigated the species boundaries by using DNA sequences of mitochondrial and nuclear molecular markers, implementing several analytical tools and species delimitation methods. The result of that approach revealed five highly divergent genetic lineages that were interpreted as independent evolutionary units, or species. Here, we present the second part of the study, where we describe three of the five species for which we have sexually mature adult specimens obtained from the mouth cavity of fish-eating birds. Additionally, we characterise morphologically the metacercariae of the other two species, collected from freshwater fishes; these species cannot be formally described since no adults were found in their definitive hosts. We further discuss the characters that are more reliable for species identification within Clinostomum, such as the cirrus sac shape and relative position with respect to testes and ovary, the shape of the reproductive organs, and the diverticulated condition of the caeca.

Specific noninvasive detection of Leishmania donovani in desquamated buccal cell swab samples from human visceral Leishmaniasis-HIV coinfected patients.

Diagnosis of human immunodeficiency virus (HIV) infection with visceral leishmaniasis (VL) coinfection is challenging. Specific diagnosis of VL in HIV-coinfected patients was evaluated by molecular methods in desquamated buccal swab samples, demonstrating 86.3% sensitivity and 98.3% specificity in controls. This test holds significant potential for development as a noninvasive diagnostic tool for VL in HIV-coinfected patients.

A case of larva migrans representing a pitfall in oral diagnosis.

The present work reports a case of a female patient complaining of itching and painful lesions affecting the oral mucosa for 7 months. Buccal and lip mucosa showed swelling and erythema, with serpiginous tracks. The patient was diagnosed with oral larva migrans, and the lesions resolved after ivermectin administration. At 18-month follow-up, no sign of recurrence was observed. Larva migrans can represent a pitfall in oral diagnosis and a stressful condition for the patient. Oral health care providers should be aware of this and keep this disease in mind as a possible differential diagnosis in oral mucosa lesions.

Importance of the CCR5-CCL5 axis for mucosal Trypanosoma cruzi protection and B cell activation.

Trypanosoma cruzi is an intracellular parasite and the causative agent of Chagas disease. Previous work has shown that the chemokine receptor CCR5 plays a role in systemic T. cruzi protection. We evaluated the importance of CCR5 and CCL5 for mucosal protection against natural oral and conjunctival T. cruzi challenges. T. cruzi-immune CCR5(-/-) and wild-type C57BL/6 mice were generated by repeated infectious challenges with T. cruzi. CCR5(-/-) and wild-type mice developed equivalent levels of cellular, humoral, and protective mucosal responses. However, CCR5(-/-)-immune mice produced increased levels of CCL5 in protected gastric tissues, suggesting compensatory signaling through additional receptors. Neutralization of CCL5 in CCR5(-/-)-immune mice resulted in decreased mucosal inflammatory responses, reduced T. cruzi-specific Ab-secreting cells, and significantly less mucosal T. cruzi protection, confirming an important role for CCL5 in optimal immune control of T. cruzi replication at the point of initial mucosal invasion. To investigate further the mechanism responsible for mucosal protection mediated by CCL5-CCR5 signaling, we evaluated the effects of CCL5 on B cells. CCL5 enhanced proliferation and IgM secretion in highly purified B cells triggered by suboptimal doses of LPS. In addition, neutralization of endogenous CCL5 inhibited B cell proliferation and IgM secretion during stimulation of highly purified B cells, indicating that B cell production of CCL5 has important autocrine effects. These findings demonstrate direct effects of CCL5 on B cells, with significant implications for the development of mucosal adjuvants, and further suggest that CCL5 may be important as a general B cell coactivator.

Post-kala-azar dermal leishmaniasis with mucosal involvement: an unusual case presentation including successful treatment with miltefosine.

Post-kala-azar dermal leishmaniasis (PKDL) is a dermatologic manifestation that usually occurs after visceral leishmaniasis (VL) caused by Leishmania donovani. It is characterized by hypopigmented patches, a macular or maculopapular rash and nodular skin lesions on the body surface. Involvement of the mucosae is very rare and unusual in PKDL. We report a case of PKDL that presented with polymorphic skin lesions, along with involvement of peri-oral mucosa and tongue from an endemic area for kala-azar in Bangladesh. In the absence of a definite past history of kala-azar, a clinical suspicion for PKDL was confirmed by positive rapid serological tests against two recombinant (rK39 and rK28) leishmanial antigens, demonstration of Leishmania donovani (LD) body in the slit skin smear, and isolation of promastigotes by culture from a nodular lesion. The patient was treated with oral Miltefosine for three consecutive months and showed significant clinical improvement as demonstrated by a negative slit skin smear at two months after initiation of therapy. We report this case as an unusual presentation of mucosal involvement in PKDL and subsequent treatment success with Miltefosine.

Mixed mucosal leishmaniasis infection caused by Leishmania tropica and Leishmania major.

Mixed infections with different Leishmania species could explain differences in the clinical courses of these infections. On identification of Leishmania parasites from Iranian patients with mucosal leishmaniasis (ML), a patient with both oral and nasal lesions was found to be concomitantly infected with Leishmania tropica and L. major. Mixed infection was identified by PCR amplification of Leishmania kinetoplast DNA on scraping of cytological smears and histopathological sections. L. major and L. tropica were isolated from the nasal and oral lesions, respectively. These species were also confirmed by immunohistochemistry. This seems to be the first reported case of concurrent ML infection with two Leishmania species. It indicates that, at least in this patient, previous infection with one of these Leishmania species did not protect against infection with the other. This result has important implications for the development of vaccines against leishmaniases and implies careful attention in the treatment of this infectious disease.

Molecular diagnosis and species identification of mucosal leishmaniasis in Iran and correlation with cytological findings.

OBJECTIVE: Mucosal leishmaniasis (ML) is a rare destructive disease that mainly affects the mucous membranes of the mouth and nose. The etiologic agent(s) of ML are not well known in the Middle East. STUDY DESIGN: Cytologic smears of ML from the mucosal lesions of 7 patients were prepared by scraping. In 2 patients with nasal lesions, exfoliative cytology was made by washing the nasal cavity. The smears were both air dried and fixed in alcohol and stained. Scrapings from the same smears were then tested for leishmanial DNA by nested PCR. RESULTS: This study characterized 9 isolates of ML, with 7 cases identified as Leishmania major and 2 as Leishmania tropica. While 6 patients were found to be positive by the cytology technique, the nested PCR was positive for all of these samples. CONCLUSIONS: Presence of granuloma and multinucleated giant cells in the negative smears of the patients who showed clinical manifestation of ML was an important clue for diagnosis of this disease. The PCR-based method not only appears to be a precise diagnostic approach in the identification of suspected cases of ML but is also efficient in determining the species of the parasite. L. major and L. tropica can lead to ML, but they result in different cytologic features.

Oral myiasis in an elderly patient.

BACKGROUND: Myiasis is the invasion of living tissue of humans and other mammals by eggs or maggots of flies of the order of Diptera. It occurs mainly in the Tropics and is associated with inadequate public and personal hygiene. Oral myiases in an older man appears to be rare. OBJECTIVE: To relate a case of oral myiases in a debilitated older man treated by mechanical removal of the maggots, identifying the adult insect that caused the infestation. METHODS: The diagnosis of oral myiasis was established by the clinical examination and it was detected that the infestation involved only soft tissue and the sinus cavity. The patient was submitted to two mechanical removal of the visible maggots. RESULTS: Total of 110 maggots was removed from the oral cavity of the patient and adult insects was identified as belonging to the Calliphoridae Family, Cochliomyia hominivorax species. The patient died two days after the second procedure by severe systemic complications. CONCLUSIONS: The mechanic removal and the identification of the maggots must be adopted as soon as possible to prevent further tissue damage and bacterial infection in cases of oral myiasis. Special attention should be given to the debilitated old patients that are particularly susceptible to oral myiasis infestation.

Social exclusion and myiasis of the upper lip in a Brazilian rural resident.

CONTEXT: People living in rural or remote Brazil, as in other sub-tropical or tropical nations, are vulnerable to infections that would not normally occur in urban areas or wealthier nations. Brazil is a geographically extensive nation, historically marked by vast socioeconomic inequalities. Approximately 16% of the population live in rural areas. ISSUE: This clinical case report demonstrates the vulnerability of rural residents who are underserved by Brazil's 'universal' public healthcare system, despite social and economic challenges that increase their risk for disease. Myiasis (especially oral myiasis) is a rare health condition in humans caused by fly larvae. Oral myiasis usually appears in periodontal pockets and open wounds, such as after dental extractions. It is associated with poverty, lack of access to health care, and very poor overall health status. While myiasis has a worldwide distribution, it is particularly associated with the tropical and sub-tropical regions of North and South America. LESSONS LEARNED: This article describes a rare case of myiasis in the upper lip of a rural male patient. The case report demonstrate that rural and remote residents can be socially excluded from the benefits of technology and biomedicine, making them vulnerable to rare infections.

Noninvasive molecular diagnosis of human visceral leishmaniasis.

Previously developed methods for noninvasive PCR diagnosis of visceral leishmaniasis (VL) have significant limitations. Diagnosis of VL using PCR and buccal swabs was evaluated in 307 subjects, including 148 patients confirmed to have VL. This method is simple and well tolerated and has good potential for development, showing 83% sensitivity with 90.56% specificity in control groups.

High prevalence of buccal ulcerations in largemouth bass, Micropterus salmoides (Centrarchidae) from Michigan inland lakes associated with Myzobdella lugubris Leidy 1851 (Annelida: Hirudinea).

Widespread mouth ulcerations were observed in largemouth bass collected from eight inland lakes in the Lower Peninsula of Michigan during the summer months of 2002 and 2003. These ulcerations were associated with, and most likely caused by, leech parasitism. Through the use of morphological dichotomous keys, it was determined that all leeches collected are of one species: Myzobdella lugubris. Among the eight lakes examined, Lake Orion and Devils Lake had the highest prevalence of leech parasitism (34% and 29%, respectively) and mouth ulcerations (53% and 68%, respectively). Statistical analyses demonstrated that leech and ulcer prevalence varied significantly from one lake to the other. Additionally, it was determined that the relationship between the prevalence of ulcers and the prevalence of leech attachment is significant, indicating that leech parasitism is most likely the cause of ulceration. The ulcers exhibited deep hemorrhagic centers and raised irregular edges. Affected areas lost their epithelial lining and submucosa, with masses of bacteria colonizing the damaged tissues. Since largemouth bass is a popular global sportfish and critical to the food web of inland lakes, there are concerns that the presence of leeches, damaged buccal mucosa, and general unsightliness may negatively affect this important sportfishery.

Primary oral myiasis due to Chrysomya bezziana treated with Ivermectin. A case report.

Primary oral myiasis due to Chrysomya bezziana is a rare condition caused by invasion of tissues by larvae of the flies. A case of Oral myiasis is presented in a 12 year old boy with neuro-degenerative disease with seizures. Intra orally, a soft tissue pocket in the left buccal mucosa and a pocket under the palatal mucosa was seen containing maggots. Extra orally indurated erythematous swelling was present near the commissure of the mouth. Predisposing factors identified in the present case were mouth breathing, incompetent lips, low socioeconomic condition, malnutrition, and inability of the child to perform daily activities due to his neurodegenerative disease. Treatment consisted of manual removal of maggots following irrigation and application of turpentine oil along with a single dose Ivermectin 3 mg that was given systemically. Complete healing of the lesion was observed within 10 days without any recurrence for a follow up period of 1 year Medical personnel taking care of old / debilitated / unconscious patients need to bear in mind the possibility of Chrysomya bezziana infestation in them.

Oral myiasis caused by Cochliomyia hominivorax treated with ivermectin and nitrofurazone: case report.

Myiasis is the infestation of tissues and organs of animals and humans by certain dipteran fly larvae. This phenomenon is well-documented, especially among animals and people in tropical and subtropical areas. Oral myiasis is a rare condition that can be caused by several species of dipteran fly larvae. When the larvae are removed from the mouth, the tissues seem to recover with no subsequent complications and no need for further treatment. This article presents a case involving a patient with oral myiasis caused by Cochliomyia hominivorax and treated with ivermectin and nitrofurazone.

Oral myiasis in a captive hippopotamus.

Causes of dental infections can be related to failed dental eruption, malocclusion, abrasion, fractures with or without exposure of the dental pulp, and periodontal disease. Reports of oral myiasis in megavertebrates in captivity are infrequent, perhaps due to the difficulty in observing the oral cavity in such species. This report describes a case of oral myiasis in an adult male hippopotamus in the gingival area and alveolar mucosa of the left mandibular canine tooth.

Human dirofilariosis of buccal mucosa - First molecularly confirmed case and literature review.

When presented in unusual manner, human dirofilariosis is hard to recognize and often leads to misdiagnosis and unjustified use of various medications. Herein, we report a rare case of Dirofilaria infection localized in buccal mucosa. A 45-year-old male, residing in the Central Serbia, developed striking edema of buccal mucosa followed by leucocytosis, with both neutrophylia and eosinophylia. Thirty days after first symptoms occurred, nodule was formed along the lateral edge of right maxilla. Therefore, patient underwent surgical extirpation and subsequent histopathological analysis of the specimen revealed the presence of a nematode. Based on morphological characteristics of detected parasite, the diagnosis of submucosal Dirofilaria repens-like infection was made. The diagnosis was confirmed, being the first case in the world analyzed by molecular methods. So far, only 13 such cases have been reported worldwide and our paper brings attention to rare form of this zoonosis and updates the data about human dirofilariosis in buccal mucosa.

Fulminant presentation of oral mucosal leishmaniasis as severe stomatitis and periodontitis.

This case report shows an atypical presentation of mucosal leishmaniasis infantum in the oral cavity resulting in severe stomatitis and periodontitis. The patient was immunocompromised because of rheumatoid arthritis for which he used prednisone and methotrexate. He was treated with intravenous liposomal amphotericin B and recovered within four weeks.