Epidemiological study of sporotrichosis and histoplasmosis in captive Latin American wild mammals,São Paulo,Brazil

Sporotrichosis and histoplasmosis are deep mycosis with a high incidence in human beings in Brazil. In domestic animals histoplasmosis has been described only in dogs, but the occurence of sporotrichosis among domestic animals in Brazil has been described in dogs, cats, mules and asses. There is also a case of this disease reported in a chimpanzee (Pan troglodites). The purpose of this research was to perform an epidomiological study of these mycoses using delayed hypersensitivity tests (histoplasmin and sporotrichin) in Latin American wild mammals. This research was assayed using 96 healthy animals at Parque Zoológico de São Paulo, Brazil: Primates: 33Cebus apella — weeping-capuchin and 16Callithrix jacchus — marmoset; Procyonidae: 37Nasua nasua — coatimundi and 10 Felidae (Panthera onca — jaguar;Felis pardalis — ocelotFelis wiedii — margay;Felis tigrina — wild cat). For intradermic tests, the following antigens were used:Sporothrix schenkii cell suspension (sporotrichin, histoplasmin-filtrate),Histoplasma capsulatum cell suspension (histoplasmin), andHistoplasma capsulatum (polysaccharide). The positivity to histoplasmin was 44.79% (Cebidae 15.15%; Callithricidae 6.25%; Procyonidae 86.49% and Felidae 50.00%, respectively). With respect to sporotrichin, 30.21% (Cebidae 6.06%, Callithricidae 0.0%; Procyonidae 64.86% and Felidae 30.00% respectively). The pattern of infection is similar to that shown by human beings and this may suggest that these animals could be involved in the epidemiologic chain of sporotrichosis and histoplasmosis, the second most prevalent human deep mycoses in Brazil. It is important to point out the absence of similar studies in Latin American wild animals.     

The status of histoplasmosis in Brazil

Thirty four cases of the progressive form of histoplasmosis, five epidemics of the mycosis, many histoplasmin skin test surveys and few clinical or histological evidences of the benign form of the disease were reported in Brazil. The occurrence of the infection in animals and the isolation ofH. capsulatum from soil have also been reported.Pesquisador 1 B do CNPq.     

Additional studies of histoplasmin formation

Summary Culture filtrates of 20 strains ofHistoplasma capsulatum were studied to determine the effect of certain growth conditions on histoplasmin formation. The presence of histoplasmin was denoted by an antigenic titer of 1:4 or higher with the complement fixation test.The data indicated that, in addition to verifying that the strain used affected histoplasmin formation, the morphological condition of the inoculum was extremely important. It was found that most strains which converted readily to the yeast phase at 37° C produced histoplasmin poorly. Tests with different volumes of media also showed that 500 ml volumes of culture media produced histoplasmin with higher titers than 3 liter volumes when cultured at 25° C for six months.Some additional histoplasmin could be liberated by sonification of the mycelial pad from culture filtrates which contained histoplasmin. A few strains produced high titer histoplasmin by the shake method if incubated for three months, but they had low titers after only six weeks.Complement fixation tests with sera from proven cases of histoplasmosis indicated that histoplasmin from a single strain ofH. capsulatum can give identical results with those obtained with histoplasmin from a pool ofH. capsulatum strains if H and M antigen components are present.     

A preliminary note on inter-reaction of skin test sensitivity between Histoplasmin and Chrysosporin in experimental animals

Summary Species ofChrysosporium have been isolated from soil in Iran. Guinea pigs inoculated withChrysosporium keratinophilum gave a positive skin test to histoplasmin and vice versa. This would suggest thatChrysosporium species might be reponsible for positive skin tests to histoplasmin in areas known not to be endemic for histoplasmosis.     

Epidemiologic skin test survey of sensitivity to paracoccidioidin,histoplasmin and sporotrichin among gold mine workers of Morro Velho Mining,Brazil

Skin tests with paracoccidioidin, histoplasmin and sporotrichin were applied to 417 workers of Morro Velho Mining in the State of Minas Gerais, Brazil, with the main purpose of detecting the prevalence of paracoccidioidomycosis-infection, histoplasmosis capsulate-infection and sporotrichosis-infection. The rates of positivity to the skin tests were 13.43% for paracoccidioidin, 17.50% for histoplasmin and 13.67% for sporotrichin. Several epidemiological factors were investigated for a better interpretation of the results. Paracoccidioides brasiliensis, Histoplasma capsulatum var. capsulatum and Sporothrix schenkii were not isolated from the soil samples from the mines investigated.     

An unusual presentation of disseminated histoplasmosis in a non-HIV patient from Vietnam

BackgroundHistoplasmosis is a systemic infection caused by the dimorphic fungus Histoplasma capsulatum, naturally found in nitrogen-rich soil, whose main transmission route is the inhalation of conidia. Up to 95% of histoplasmosis cases are asymptomatic or transient, and the remaining 5% of cases have pathological manifestations in the lungs, bone marrow, liver, spleen, intestine, mucous membranes, and rarely on the skin. This mycosis has been reported from many endemic areas, mainly in immunosuppressed patients, such as HIV-positive patients, and its disseminated form is rarely reported.Case reportHistoplama capsulatum was isolated and identified by means of microscopy, culture characteristics and nested PCR from the cutaneous lesions of a non-HIV patient from Vietnam. The patient improved significantly with systemic itraconazole treatment.ConclusionsDisseminated histoplasmosis with cutaneous involvement in non-HIV patients is an extremely unusual presentation.     

Histoplasmosis in Europe

After reviewing the reports ofHistoplasma capsulatum in man, animal and soil, and the results of histoplasmin sensitivity tests in Europe, we have concluded that autochthonous histoplasmosis is present but rare in Europe.Only the European parts of Turkey and of U.S.S.R. are considered in this report.Paper read at the Eighth International Congresses for Tropical Medicine and Malaria, September 1968, Teheran (Iran).     

The asteroid body of lobomycosis

The epidemiology of histoplasmosis duboisii (African histoplasmosis) is not well understood. The present study was carried out to investigate the prevalence of skin sensitivity and to determine by immunodiffusion the presence of antibodies among humans to histoplasmin around a recently discovered natural focus of Histoplasma capsulatum var. duboisii in a bat cave in Ogbunike in the Anambra State of Nigeria. Out of the 40 subjects, all young adults aged 18–30 years, comprising cave guides, traders and farmers examined in the immediate vicinity of the cave, 14 (35.0%) gave a positive skin test. In another population of the same age group, comprising 620 persons, viz. traders, farmers, palm oil workers and some patients attending rural clinics, examined in other nearby areas in Anambra State, 55 (8.8%) reacted positively to histoplasmin. In the immunodiffusion tests, 2 (2.08%) of the 96 school children and 17 (9.4%) of the 181 young adults, including farmers, palm oil workers and traders tested amongst the population around the cave, demonstrated precipitating antibodies to histoplasmin in their sera. Only 5 (0.79%) of the 630 adults of the same age group with similar occupations examined from other areas in Anambra State had precipitating antibodies. Out of another 50 subjects examined, viz.; wood workers, traders, farmers, and school teachers in Nsukka in the Enugu State, two (4.0%) demonstrated antibodies. It is suggested that asymptomatic infections due to the duboisii variety of H. capsulatum may be common in the human population around the cave. A diligent search with the help of local hospitals and public health officials may reveal clinical cases of histoplasmosis duboisii with cutaneous and systemic lesions.     

Histoplasmosis in northwestern Argentina II: Prevalence ofHistoplasmosis capsulati and Paracoccidioidomycosis in the population south of Chuscha,Gonzalo and Potrero in the province of Tucuman

The present work was undertaken to obtain epidemiological data on the extent and distribution ofHistoplasma capsulatum var.capsulatum andParacoccidiodes brasiliensis infections south of the Chuscha, Gonzalo and Potrero areas of Argentina. Skin tests surveys of the human population with histoplasmin and paracoccidioidin were carried out in the permanent population of those localities. The infection index of the population showed that the area south of Chuscha has a high-prevalence of histoplasmosis capsulati. The Gonzalo and Potrero areas, according to their rates of infection also can be considered to have a relatively high prevalence of this disease. The frequency of individuals infected withP. brasiliensis suggests that the level of exposure to this fungus is considerable, especially in Gonzalo where the frequency of infection was 9.23%. The endemic areas of both diseases can be superimposed, as occurs in the northeastern part of Argentina.     

Chronic Cavitary Histoplasmosis of the Lung

Four main clinical types of disease result from infection by the fungus Histoplasma capsulatum: (1) the primary complex; (2) acute pulmonary histoplasmosis; (3) chronic cavitary pulmonary histoplasmosis; (4) disseminated systemic histoplasmosis.Chronic cavitary histoplasmosis of the lung is indistinguishable clinically and radiographically from pulmonary tuberculosis. In this disease, however, the histoplasmin skin test and the histoplasmin complement fixation test are positive in more than 80% of cases and sputum cultures are usually positive for Histoplasma capsulatum. The mortality rate of the chronic cavitary type is greater than 30%. The antibiotic, amphotericin B, is the most effective drug in treatment. If surgical resection of the cavitary lesion is feasible, under amphotericin coverage, this is the treatment of choice.During the past year, two patients with chronic cavitary histoplasmosis illustrating the aforementioned features were diagnosed at the Toronto Hospital for Tuberculosis.     

Immunology of histoplasmosis: Humoral and cellular activity from a polysacchari-de-protein complex and its deproteinized fraction in experimentally immunized mice

In a previous publication it was reported that a polysaccharide-protein complex (PPC), sensitive to -glucosidase, was isolated from Histoplasma capsulatum. This complex was strongly reactive in an agar gel diffusion assay with sera from patients with histoplasmosis, but was unreactive with sera from patients with coccidioidomycosis. Here, the studies with human sera have been expanded and attempts were made to determine the response of mice immunized with nonviable H. capsulatum or Cocccidioides immitis to PPC or its deproteinized fraction (D-PPC) using more sensitive tests for antibody and including also test for cell-mediated immunity. Histoplasmin and coccidioidin were compared with PPC or its deproteinized fraction (D-PPC) in all assays. In a counterimmunoelectrophoresis (CIE) assay, PPC and D-PPC reacted only with sera from patients with histoplasmosis, whereas cross reactions were noted with histoplasmin and coccidioidin using heterologous sera. Cross-reaction were observed with all four antigen preparations and both types of antisera using a micro complement fixation assay. The assay for macrophage migration inhibitory factor (MIF) was also relatively nonspecific, in that inhibition occurred with cells from animals sensitized with Histoplasma or Coccidioides using both homologous and heterologous antigens. In the footpad assay, histoplasmin and coccidioidin were highly cross-reactive in animals sensitized with the heterologous fungus, but the PPC and D-PPC from H. capsulatum elicited significant reactions only in animals sensitized with Histoplasma.     

Deep mycoses in India

Available published reports on deep mycoses in India have been critically and exhaustively reviewed. So far there seem to be only 9 cases of actinomycosis reported, mostly of thoracic type and diagnosed on the basis of the presence of sulphur granules in the lesions. Nocardiosis and its chief causal agentNocardia asteroides have received particular attention in recent studies. To-date there are 18 authentic cases reported from India and significantly 12 of these have been diagnosed by applying the paraffin bait technique to the isolation ofN. asteroides from sputa and other clinical specimens. In most of these 12 cases timely diagnosis allowed for the successful treatment of the disease with heavy doses of sulphadiazine. Case reports on cryptococcosis which include 26 adequately documented cases, have been published from various parts of the country. Occurrence ofCryptococcus neoformans in soil and its association with old pigeon excreta have also been confirmed by studies done in some northern and western regions of this country. The status of histoplasmosis in India still remains a debatable subject although there is a suggestive evidence that the disease may be endemic in the northeastern parts. There are 9 case reports in which diagnosis has been supported by histopathologic findings and in 3 cultures have also been positive. However, attempts to isolateHistoplasma capsulatum from soil or any other extra-human source have remained futile and the limited surveys have revealed only low skin sensitivity to histoplasmin and none to blastomycin and coccidioidin. As yet there is no authentic case of blastomycosis, coccidioidomycosis or paracoccidioidomycosis reported from India. Two cases of invasive aspergillosis and 6 of bronchopulmonary aspergillomas have been published. In the latterAspergillus fumigatus, A. niger andA. flavus have been found to be the aetiologic agents. In addition, a recent report on a series of 8 patients recognises for the first time the occurrence of allergic aspergillosis in this country. Two cases of phycomycoses, involving the lungs in one and brain in the other case have been described. Diagnosis of bronchopulmonary candidiasis has been claimed in as many as 16 patients by several authors but in none the evidence is unequivocal. The isolation ofCandida viswanathii from the cerebrospinal fluid of two fatal cases is suggestive of the possible aetiologic role of this new yeast in human meningitis. Besides, there are 3 cases of brain mycoses described in Indian literature, two due toCladosporium trichoides while in the third caseUstilago maydis, the causal agent of maize smut, has been implicated.     

THE DISTRIBUTION OF COCCIDIOIDOMYCOSIS IN SOUTHERN CALIFORNIA

In coccidioidin skin test surveys among persons of high school age in Saugus, Canoga Park, Banning and Palm Springs areas the average incidence of positive reaction was 15 per cent. Although considerably less than the 68 per cent incidence reported among high school students of Kern County, it is high enough to indicate pockets of relatively high endemicity in Southern California below the San Joaquin Valley.Histoplasmin tests were performed on most of the persons tested with coccidioidin in this survey. The over-all incidence of positive reaction in the group was 7.6 per cent. Most of the subjects with positive reaction to histoplasmin gave a history of having previously lived in some area in the central United States where histoplasmosis is known to be endemic.A few subjects who had positive reaction to coccidioidin tests and who had lived in areas known to be endemic for coccidioidomycosis but not for histoplasmosis, also had positive reaction to histoplasmin. However, the induration produced was always smaller than that caused by the coccidioidin reaction, and there was minimal confusion in interpreting the tests.     

Studies on histoplasmosis IV. Elution characteristics of the Histo-inhibitory factor (HIF)

Summary The histo-inhibitory factor (HIF) derived from homogenates of liver or kidney from hamsters infected withHistoplasma capsulatum has been fractionated by column chromatography. It shows maximum absorption at 280 mµ, has a molecular weight of 142,000 and can be eluted from DEAE-cellulose or DEAE-Sephadex A—50 with 0.02 M phosphate — 4 M sodium chloride (1 : 1)HIF can be eluted fromHistoplasma yeast cells at pH 10.0, thus a greater number of positive cultures from chronic histoplasmosis could be expected to result from pretreatment of clinical specimens in glycine buffer pH 10.0 prior to culture.     

Histoplasma capsulatum in the environment of sporadic histoplasmosis cases

Conclusions 1. Acute pulmonary histoplasmosis in adults is demonstrated to be associated with exogenous sources of infection in one-half of a series of cases. 2. No exogenous sources of infection were found in chronic pulmonary histoplasmosis in adults by methods comparable to those used in acute pulmonary histoplasmosis. 3. The significance of these findings for the pathogenesis of the three clinical forms of histoplasmosis is discussed. 4. In acute pulmonary histoplasmosis in adults, the isolation ofH. capsulatum from the environment at sites of exposure aided in specific diagnosis.Two acute pulmonary histoplasmosis cases were reported previously (2–3).     

Gastrointestinal disseminated histoplasmosis in HIV-infected patients: A descriptive and comparative study

Disseminated histoplasmosis is one the main AIDS-defining opportunistic infections in HIV-infected patients, notably in Latin America. The non-specific and proteiform clinical presentation leads to diagnostic delays that may lead to fatal outcomes. This retrospective multicentric study aimed to describe the frequency and manifestations of gastrointestinal histoplasmosis in French Guiana, and to compare patients with disseminated histoplasmosis with or without gastrointestinal involvement.Between January 1, 1981 and October 1, 2014 co-infections with HIV and histoplasmosis were enrolled. Inclusion criteria were: age >18 years, confirmed HIV infection; first proven episode of histoplasmosis.Among 349 cases of disseminated histoplasmosis, 245 (70%) had a gastrointestinal presentation. Half of patients with gastrointestinal signs had abdominal pain or diarrhea, mostly watery. Half of patients with abdominal pain had diarrhea (63/124) and half of those with diarrhea (63/123) had abdominal pain. A significant proportion of patients also had hepatomegaly and, to a lesser degree, splenomegaly. After adjusting for potential confounding, the presence of lymphadenopathies >2cm (AOR = 0.2, IC95 = 0.04–0.7, P = 0.01), Haitian origin (AOR = 0.04, IC95 = 0.004–0.4, P = 0.006) were associated with a lower prevalence of gastrointestinal signs and positive gastrointestinal presence of H. capsulatum. Persons with a gastrointestinal H. capsulatum were more likely to have a decreased prothrombin time, lower ferritin, lower liver enzymes, and lower concentrations of LDH than those without gastrointestinal signs and symptoms. They also had a shorter interval between symptoms onset and diagnosis. Patients with a positive gastrointestinal identification of H. capsulatum were less likely to die at 1 month than those without a gastrointestinal presentation (respectively, 4.6% vs 18.5%, P = 0.01).Subacute or chronic gastrointestinal presentations are very frequent during disseminated histoplasmosis, they seem less severe, and should lead to suspect the diagnosis in endemic areas. There were populational or geographic differences in the frequency of gastrointestinal manifestations that could not be explained.     

Epididymal histoplasmosis diagnosed by isolation ofHistoplasma capsulatum from semen

An autochthonous case of epididymal histoplasmosis masquerading as tuberculosis in a 55-year-old male patient is reported from India. It was diagnosed by culture ofHistoplasma capsulatum from semen and by demonstration of the fungus upon re-examination of epididymal biopsy sections previously misinterpreted as tuberculous granuloma. The patient's main complaints were painful epididymal swelling, occasional fever and cough. He was treated successfully by excision of epididymis and vas deferens combined with amphotericin B therapy. This is believed to be the first case of epididymal histoplasmosis to be reported outside the American continent and the fourth of its type reported in the English literature. The case is also noteworthy in thatH. capsulatum was isolated for the first time from semen, and it underlines the importance of mycological culture of semen specimens for diagnosis of genitourinary infections of obscure etiology.Presented at the XII Congress of the International Society for Human and Animal Mycology, Adelaide, Australia, March 13–18, 1994.     

A redescription ofDiaphanosoma modigliani Richard, 1894 (Crustacea : Daphniiformes : Sididae)and an evaluation of its occurence outside Lake Toba (Sumatra)

A pelagic species of Diaphanosoma, D. modigliani is redescribed on material from the type locality (Lake Toba, Sumatra, Indonesia). Other records of this species in tropical Asia (India, Philippine Islands, Malaysia, Sri Lanka, Thailand) are either unsubstantiated or erroneous.     

In situ localization of antigens of Histoplasma capsulatum using colloidal gold immune electron microscopy

Histoplasma capsulatum contains multiple antigens, among them the H antigen and M antigen, which are useful in serologic testing for histoplasmosis. We prepared 7 mouse monoclonal antibodies (5 IgG, 2 IgM) to histoplasmin, and compared these with polyclonal histoplasmin antibodies raised in rabbits and mice. Both monoclonal and polyclonal antibodies were high titered by ELISA. Colloidal gold immune electron microscopy (CGIEM) showed that polyclonal antibodies to histoplasmin or H antigen bound at multiple sites in the cell wall, cytoplasm, and nucleus of Histoplasma yeast cells. In contrast, antibodies to M antigen selectively label the cell membrane and antibodies to alkali soluble cell wall antigen label only the cell wall. Polyclonal antibodies cross reacted extensively with other fungi, both by ELISA and CGIEM. Monoclonal antibodies stained only cytoplasmic epitopes, but also cross reacted with other fungi by electron microscopy. Only periodate treated H antigen elicited polyclonal antibodies which were more specific than those of untreated H antigen or histoplasmin.     

A natural focus ofHistoplasma capsulatum var.duboisii is a bat cave

The natural reservoir ofHistoplasma capsulatum var.duboisii, the etiological agent of histoplasmosis duboisii (African histoplasmosis) is not yet known. We report the isolation ofH. capsulatum var.duboisii from soil admixed with bat guano and from the intestinal contents of a bat in a sandstone cave in a rural area, Ogbunike in Anambra State of Nigeria. Eight of 45 samples of soil admixed with bat guano yieldedH. capsulatum var.duboisii. Of the 35 bats belonging to the speciesNycteris hispida andTadirida pumila examined, only one (N. hispida) yielded this fungus from its intestinal contents. Identification of the isolates asHistoplasma was confirmed by exoantigen tests and by mating with tester strains ofH. capsulatum. In vitro conversion to large yeast from suggestive ofH. capsulatum var.duboisii was obtained on brain heart infusion agar supplemented with sheep blood and glutamine or cysteine. Pathogenicity tests with mice for all the isolates confirmed their identity by the demonstration of large yeast forms (8–15 µm in diameter) within giant cells in the infected tissues. Investigations on the possible occurrence of human infections in the area are in progress.A poster based on this work was presented at the 11th ISHAM Congress in Montreal, Canada (22–28 June 1991), La-Hoffman Roche, Basel, Switzerland kindly financed the trip of one of us (H.C.G) for the Congress.