EFFECTS OF SINGLE,BINARY AND TERTIARY COMBINATIONS WITH Jatropha gossypifolia AND OTHER PLANT-DERIVED MOLLUSCICIDES ON REPRODUCTION AND SURVIVAL OF THE SNAIL Lymnaea acuminata

The effect of sub-lethal doses (40% and 80% ofLC₅₀/24h) of plant derived molluscicides of singly,binary (1:1) and tertiary (1:1:1) combinations of the Rutin,Ellagic acid, Betulin and taraxerol with J. gossypifolia latex,leaf and stem bark powder extracts and their active component on thereproduction of freshwater snail Lymnaea acuminata have beenstudied. It was observed that the J. gossypifolia latex, stembark, individual leaf and their combinations with other plant derived activemolluscicidal components caused a significant reduction in fecundity,hatchability and survival of young snails. It is believed that sub-lethalexposure of these molluscicides on snail reproduction is a complex processinvolving more than one factor in reducing the reproductive capacity.     

SURVIVAL,INDUCTION AND RESUSCITATION OF Vibrio cholerae FROM THE VIABLE BUT NON-CULTURABLE STATE IN THE SOUTHERN CARIBBEAN SEA

The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.     

A PRELIMINARY STUDY ON THE OCCURRENCE OF KERATINOPHILIC FUNGI IN SOILS OF JAMAICA

This report represents the first study of keratinophilic fungi present insoils of Jamaica. Out of the 40 soil samples examined from different habitats, 30(75%) were positive for the presence of keratinophilic fungi, yielding 36 isolates ofkeratinophilic fungi. Microsporum gypseum complex (represented by 16isolates of M. gypseum, and four of M. fulvum) wasmost frequent, being present in 50% of the samples. A very high occurrence of thisdermatophyte in Jamaican soil is of public health significance. The remainingisolates of keratinophilic fungi were represented by Chrysosporiumspp (mainly C. indicum and C. tropicum) andSepedonium sp.     

Anopheles (Kerteszia) cruzii (DIPTERA: CULICIDAE) IN PERIDOMICILIARY AREA DURING ASYMPTOMATIC MALARIA TRANSMISSION IN THE ATLANTIC FOREST: MOLECULAR IDENTIFICATION OF BLOOD-MEAL SOURCES INDICATES HUMANS AS PRIMARY INTERMEDIATE HOSTS

Anopheles (Kerteszia) cruzii has beenimplicated as the primary vector of human and simian malarias out of theBrazilian Amazon and specifically in the Atlantic Forest regions. The presenceof asymptomatic human cases, parasite-positive wild monkeys and the similaritybetween the parasites infecting them support the discussion whether theseinfections can be considered as a zoonosis. Although many aspects of the biologyof An. cruzii have already been addressed, studies conductedduring outbreaks of malaria transmission, aiming at the analysis of bloodfeeding and infectivity, are missing in the Atlantic Forest. This study wasconducted in the location of Palestina, Juquitiba, where annually the majorityof autochthonous human cases are notified in the Atlantic Forest of the state ofSão Paulo. Peridomiciliary sites were selected for collection of mosquitoesin a perimeter of up to 100 m around the residences of human malaria cases. Themosquitoes were analyzed with the purpose of molecular identification ofblood-meal sources and to examine the prevalence of Plasmodium.A total of 13,441 females of An. (Ker.) cruzii werecollected. The minimum infection rate was calculated at 0.03% and0.01%, respectively, for P. vivax and P.malariae and only human blood was detected in the blood-fedmosquitoes analyzed. This data reinforce the hypothesis that asymptomatic humancarriers are the main source of anopheline infection in the peridomiciliaryarea, making the probability of zoonotic transmission less likely to happen.     

EXPERIMENTAL INFECTION WITH Toxocara cati IN PIGS: MIGRATORY PATTERN AND PATHOLOGICAL RESPONSE IN EARLY PHASE

Experimental inoculations of approximately 100,000 infectiveToxocara cati larval eggs were done in twelve pigs. TheT. cati eggs used for inoculation were collected from cat''sfeces. Another group of three pigs served as an uninfected control. Groups ofinfected pigs were euthanized at seven, 14, 21, and 28 days post-inoculation (dpi).Tissue samples were taken for digestion and histopathology changes in early phase.The number of larvae recovered from the lungs peaked at seven and 14 dpi and werealso present at 21, and 28 dpi. Larvae of T. cati were present inthe lymph nodes of the small and large intestine at seven, 14, and 28 dpi and atseven, 14, 21, and 28 dpi respectively. In other studied tissues, no larvae or lessthan one larva per gram was detected. The pathological response observed in the liverand lungs at seven and 14 dpi, showed white spots on the liver surface and areas ofconsolidation were observed in the lungs. The lungs showed an inflammatory reactionwith larvae in center at 28 dpi. In the liver we observed periportal and perilobularhepatitis. The lymph nodes of the intestines displayed eosinophil lymphadenitis withreactive centers containing parasitic forms in some of them. The granulomatousreaction was not observed in any tissues. The role of the other examined tissues hadless significance. The relevance of this parasite as an etiological agent that leadsto disease in paratenic hosts is evident.     

DETECTION OF Leishmania (Viannia) IN Nyssomyia neivai AND Nyssomyia whitmani BY MULTIPLEX POLYMERASE CHAIN REACTION, IN SOUTHERN BRAZIL

Sandflies transmit pathogens of leishmaniasis. The natural infectionof sandflies by Leishmania (Viannia) was assessed inmunicipalities, in the state of Paraná, in Southern Brazil. Sandflies werecollected with Falcão and Shannon traps. After dissection in search offlagellates in digestive tubes and identification of the species, femalesandflies were submitted to the Multiplex Polymerase Chain Reaction(multiplex PCR) for detection of the fragment of the kDNA ofLeishmania (Viannia) and the fragment from theIVS6 cacophony gene region of the phlebotomine insects. Theanalysis was performed in pools containing seven to 12 guts from females of thesame species. A total of 510 female sandflies were analyzed, including nineMigonemyia migonei, 17 Pintomyia fischeri,216 Nyssomyia neivai, and 268 Nyssomyiawhitmani. Although none of the females was found naturally infectedby flagellates through dissection, the fragment of DNA from Leishmania(Viannia) was shown by multiplex PCR in one sample ofNy. neivai (0.46%) and three samples ofNy. whitmani (1.12%). It was concluded thatNy. neivai and Ny. whitmani aresusceptible to Leishmania infection, and that multiplex PCR canbe used in epidemiological studies to detect the natural infection of thesandfly vector, because of its sensitivity, specificity and feasibility.     

PREVALENCE OF Calodium hepaticum (SYN. Capillaria hepatica) IN Rattus norvegicus IN THE URBAN AREA OF RIO DE JANEIRO, BRAZIL

The nematode Calodium hepaticum (syn.Capillaria hepatica) is a zoonotic helminth foundmainly infecting rats. It was studied the prevalence of C.hepaticum infection in Rattus norvegicus in anurban area of Rio de Janeiro (Brazil), with low urban planning andsanitation. The presence of C. hepaticum was identified throughvisible yellowish-white lesions in liver tissue and histological analyses. Thetotal prevalence of infection was 45%, with no significant differencesbetween sex and age. The presence of infected rodents near the peridomestic areaposes substantial risk to human health.     

RAW TROPICAL OYSTERS AS VEHICLES FOR MULTIDRUG-RESISTANT Vibrio parahaemolyticus

The following study aimed to determine the antimicrobial susceptibility profile ofVibrio parahaemolyticus strains from fresh and frozen oystersCrassostrea rhizophorae sold in Fortaleza-Brazil. An antibiogramwas performed on 87 isolates using nine antibiotics: gentamicin (Gen 10 µg),ampicillin (Amp 10 µg), penicillin G (Pen 10U), ciprofloxacin (Cip 5 µg),chloramphenicol (Chl 30 µg), nalidixic acid (Nal 30 µg), tetracycline (Tet 30 µg),vancomycin (Van 30 µg) and erythromycin (Ery 15 µg). All strains were resistant to atleast one antibiotic, and 85 (97.7%) were multi-resistant, with predominance of theVan+ Pen+Amp resistance profile (n = 46). Plasmid resistance to Pen, Amp and Ery wasdetected. Thus, the risk that raw oyster consumption poses to the health of consumersis highlighted, due to the fact that these bivalves may host antibacterial-resistantmicroorganisms.     

PREVALENCE OF Entamoeba histolytica/Entamoeba dispar IN THE CITY OF CAMPINA GRANDE,IN NORTHEASTERN BRAZIL

There is a clear need to perform epidemiological studies to find the true prevalence of Entamoeba histolytica around the world. The evaluation of this prevalence has been hindered by the existence of two different species which are morphologically identical, but genetically different, namely E. histolytica, which causes amebiasis, and E. dispar, which is non-pathogenic. In Brazil, the E. dispar has been detected in communities in the Southeastern (SE) and Northeastern (NE) regions with poor sanitation. However, individuals infected with E. histolytica have been identified in other regions. There is an absence of reports on the prevalence of these parasites in the state of Paraíba, which also has areas with poor sanitary conditions where a high prevalence of the E. histolytica/E. dispar complex has been detected in children from urban slums. The present study evaluated the prevalence of E. histolytica and E. dispar in 1,195 asymptomatic children between two and 10 years of age, living in a sprawling urban slum in Campina Grande, in the state of Paraíba, in Northeastern Brazil. These children were examined and their feces samples were analyzed microscopically. A total of 553 children tested positive for the E. histolytica/E. dispar complex, and 456 of the positive samples were tested with the E. histolytica II® ELISA kit. All 456 samples were negative for the presence of the adhesin E. histolytica specific antigen. The evidence suggests that in this community E. histolytica is absent and E. dispar is the dominant species.     

ANTHELMINTIC ACTIVITY OF LAPACHOL, β-LAPACHONE AND ITS DERIVATIVES AGAINST Toxocara canis LARVAE

Anthelmintics used for intestinal helminthiasis treatment are generally effective;however, their effectiveness in tissue parasitosis (i.e. visceral toxocariasis) ismoderate. The aim of this study was to evaluate the in vitroactivityof lapachol, β-lapachone and phenazines in relation to the viability ofToxocara canis larvae. A concentration of 2 mg/mL (in duplicate)of the compounds was tested using microculture plates containing Toxocaracanis larvae in an RPMI-1640 environment, incubated at 37 °C in 5%CO₂ tension for 48 hours. In the 2 mg/mL concentration, fourphenazines, lapachol and three of its derivatives presented a larvicide/larvistaticactivity of 100%. Then, the minimum larvicide/larvistatic concentration (MLC) testwas conducted. The compounds that presented the best results were nor-lapachol (MLC,1 mg/mL), lapachol (MLC 0.5 mg/mL), β-lapachone, and β-C-allyl-lawsone (MLC, 0.25mg/mL). The larvae exposed to the compounds, at best MLC with 100% invitro activity larvicide, were inoculated into healthy BALB/c mice andwere not capable of causing infection, confirming the larvicide potential invitro of these compounds.     

IDENTIFICATION OF Pseudomonas spp. AS AMOEBA-RESISTANT MICROORGANISMS IN ISOLATES OF Acanthamoeba

Acanthamoeba is a “Trojan horse” of the microbial world. The aim ofthis study was to identify the presence of Pseudomonas as anamoeba-resistant microorganism in 12 isolates of Acanthamoeba. Allisolates showed the genus Pseudomonas spp. as amoeba-resistantmicroorganisms. Thus, one can see that the Acanthamoeba isolatesstudied are hosts of Pseudomonas.     

USE OF MUELLER-HINTON BROTH AND AGAR IN THE GERM TUBE TEST

Candida albicans is often isolated from clinical samples, thus its presumptive differentiation from other species of the same genus can be based on its ability to form the germ tube in human serum. Nevertheless, there are two other species that share this characteristic: C. dubliniensis and C. africana. The aim of this study was to compare four different substrates to perform the germ tube (GT) test. The Candida spp. isolates were identified using a manual system (135 C. albicans, 24 C. tropicalis and one C. dubliniensis). The germ tube test was performed with fresh, previously frozen serum and Mueller-Hinton (MH) broth and agar. GT was observed in 96% (130/136) of the isolates through the fresh serum technique, 94% (128/136) through previously frozen serum, 92% (125/136) in MH agar, and 90% (122/136) in MH broth. The sensitivity of each test was higher than 90%, with 100% specificity. Both the MH agar and broth were able to identify the true positives, and false positives were not found. However, some C. albicans isolates were not identified. MH agar and broth may be used in laboratory for the rapid presumptive identification of C. albicans, as an alternative method for germ tube test.     

TWO NEW RECORDS OF Isomyia paurogonita FANG AND FAN, 1986 AND Sumatria latifrons Malloch, 1926 (DIPTERA: CALLIPHORIDAE) FROM NORTHERN THAILAND,WITH REVISED KEY TO THE SPECIES OF Isomyia

During the annual fly survey at Doi Nang Kaew in Doi Saket District,Chiang Mai Province of Thailand in 2011, Isomyia paurogonita Fang& Fan, 1986 (Diptera: Calliphoridae) and Sumatria latifronsMalloch, 1926 (Diptera: Calliphoridae) were collected for the first time in Thailand.They are the rare species of the subfamily Rhiniinae (tribe Cosminini). Prior to thisfinding, fifteen species of Isomyia and two species ofSumatria were recorded from Thailand. Therefore, 96 blow flyspecies have been found in this country. These new locality records of both flies arevery important for further research on their biology and ecology in Thailand.     

SEROLOGICAL AND MOLECULAR SURVEY OF Leptospira spp. AMONG CART HORSES FROM AN ENDEMIC AREA OF HUMAN LEPTOSPIROSIS IN CURITIBA,SOUTHERN BRAZIL

Introduction: Cart horses are a re-emerging populationemployed to carry recyclable material in cities.Methods: Sixty-two horses were sampled in an endemic areaof human leptospirosis. The microscopic agglutination test (MAT) andreal-time polymerase chain reaction (qPCR) were performed.Results: A seropositivity of 75.8% with serovarIcterohaemorrhagiae in 80.8% of the horses was observed. Blood and urinewere qPCR negative. MAT showed positive correlations with rainfall(p = 0.02) and flooding(p = 0.03).Conclusions: Although horses may be constantly exposed toLeptospira spp. in the environment mostly because ofrainfall and flooding, no leptospiremia or leptospiruria were observed in thisstudy.     

DETECTION OF Leptospira spp. AND Brucella abortus ANTIBODIES IN FREE-LIVING JAGUARS (Panthera onca) IN TWO PROTECTED AREAS OF NORTHERN PANTANAL,BRAZIL

 This study aimed to assess the exposure of free-living jaguars (Panthera onca) to Leptospira spp. and Brucella abortus in two conservation units in the Pantanal of Mato Grosso, Brazil. The presence of antibodies in blood samples of eleven jaguars was investigated using autochthonous antigens isolated in Brazil added to reference antigen collection applied to diagnosis of leptospirosis by Microscopic Agglutination Test (MAT). The Rose Bengal test was applied for B. abortus antibodies. Two (18.2%) jaguars were seroreactive for the Leptospira spp. antigen and the serovar considered as most infective in both animals was a Brazilian isolate of serovar Canicola (L01). All jaguars were seronegative for B. abortus. These data indicate that the inclusion of autochthonous antigens in serological studies can significantly increase the number of reactive animals, as well as modify the epidemiological profile of Leptospira spp. infection.     

Smqnr VARIANTS IN CLINICAL ISOLATES OF Stenotrophomonas maltophilia IN BRAZIL

SUMMARY

Stenotrophomonas maltophilia contains a novel chromosomally-encoded qnr gene named Smqnr that contributes to low intrinsic resistance to quinolone. We described Smqnr in 13 clinical isolates of S. maltophilia from two Brazilian hospitals, over a 2-year period. The strains were identified by API 20 NE (bioMérieux, France). Susceptibility by microdilution method to trimetroprim/sulfamethoxazole, ciprofloxacin, levofloxacin, minocycline, ceftazidime, chloramphenicol and ticarcillin/clavulanate was performed according to CLSI. PCR detection of Smqnr gene was carried out. The sequence of Smqnr was compared with those deposited in GenBank. Pulsed-field gel electrophoresis (PFGE) of all strains was performed. Thirteen Smqnr positives isolates were sequenced and three novel variants of Smqnr were identified. All 13 Smqnr isolates had distinguishable patterns by PFGE. This is the first report of Smqnr in S. maltophilia isolated in Brazil.     

ANIMAL MODELS FOR THE STUDY OF LEISHMANIASIS IMMUNOLOGY

Leishmaniasis remains a major public health problem worldwide and isclassified as Category I by the TDR/WHO, mainly due to the absence of control. Manyexperimental models like rodents, dogs and monkeys have been developed, each withspecific features, in order to characterize the immune response toLeishmania species, but none reproduces the pathology observed inhuman disease. Conflicting data may arise in part because different parasite strainsor species are being examined, different tissue targets (mice footpad, ear, or baseof tail) are being infected, and different numbers (“low” 1×10² and “high”1×10⁶) of metacyclic promastigotes have been inoculated. Recently, newapproaches have been proposed to provide more meaningful data regarding the hostresponse and pathogenesis that parallels human disease. The use of sand fly salivaand low numbers of parasites in experimental infections has led to mimic naturaltransmission and find new molecules and immune mechanisms which should be consideredwhen designing vaccines and control strategies. Moreover, the use of wild rodents asexperimental models has been proposed as a good alternative for studying thehost-pathogen relationships and for testing candidate vaccines. To date, usingnatural reservoirs to study Leishmania infection has beenchallenging because immunologic reagents for use in wild rodents are lacking. Thisreview discusses the principal immunological findings againstLeishmania infection in different animal models highlighting theimportance of using experimental conditions similar to natural transmission andreservoir species as experimental models to study the immunopathology of thedisease.     

THE EFFECT OF HOOKWORM INFECTION AND URINARY SCHISTOSOMIASIS ON BLOOD HEMOGLOBIN CONCENTRATION OF SCHOOLCHILDREN LIVING IN NORTHERN MOZAMBIQUE

This study aims to assess the association between schistosomiasis and hookworm infection with hemoglobin levels of schoolchildren in northern Mozambique. Through a cross-sectional survey, 1,015 children from five to 12 years old in the provinces of Nampula, Cabo Delgado and Niassa were studied. Hookworm infection and urinary schistosomiasis were diagnosed, through Ritchie and filtration methods, with a prevalence of 31.3% and 59.1%, respectively. Hemoglobin levels were obtained with a portable photometer (Hemocue^®). The average hemoglobin concentration was 10.8 ± 1.42 g/dL, and 62.1% of the children presented levels below 11.5 g/dL, of which 11.8% of the total number of children had hemoglobin levels below 9 g/dL. A multiple linear regression analysis demonstrated negative interactions between hemoglobin levels and ancylostomiasis, this being restricted to the province of Cabo Delgado (β = -0.55; p < 0.001) where an independent interaction between hemoglobin levels and urinary schistosomiasis was also observed (β = -0.35; p = 0.016). The logistical regression model indicated that hookworm infection represents a predictor of mild (OR = 1.87; 95% CI = 1.17-3.00) and moderate/severe anemia (OR = 2.71; 95% CI = 1.50 - 4.89). We concluded that, in the province of Cabo Delgado, hookworm and Schistosoma haematobium infections negatively influence hemoglobin levels in schoolchildren. Periodical deworming should be considered in the region. Health education and improvements in sanitary infrastructure could achieve long-term and sustainable reductions in soil-transmitted helminthiases and schistosomiasis prevalence rates.     

EVALUATION OF METHODS IN DETECTING VANCOMYCIN MIC AMONG MRSA ISOLATES AND THE CHANGES IN ACCURACY RELATED TO DIFFERENT MIC VALUES

INTRODUCTION: Methicillin-ResistantStaphylococcus aureus (MRSA) presenting reducedsusceptibility to vancomycin has been associated to therapeutic failure. Somemethods used by clinical laboratories may not be sufficiently accurate to detectthis phenotype, compromising results and the outcome of the patient.OBJECTIVES: To evaluate the performance of methods in thedetection of vancomycin MIC values among clinical isolates of MRSA.MATERIAL AND METHODS: The Vancomycin Minimal InhibitoryConcentration was determined for 75 MRSA isolates from inpatients of Mãe deDeus Hospital, Porto Alegre, Brazil. The broth microdilution (BM) wasused as the gold-standard technique, as well as the following methods:E-test^® strips (BioMérieux),M.I.C.E^® strips (Oxoid), PROBAC^®commercial panel and the automated system MicroScan^®(Siemens). Besides, the agar screening test was carried out with 3µg/mL of vancomycin.RESULTS: All isolates presented MIC ≤ 2µg/mL for BM. E-test^® had higher concordance(40%) in terms of global agreement with the gold standard, andthere was not statistical difference among E-test^® and brothmicrodilution results. PROBAC^® panels presented MICs, ingeneral, lower than the gold-standard panels (58.66% majorerrors), while M.I.C.E.^® MICs were higher (67.99%minor errors).CONCLUSIONS: For the population of MRSA in question,E-test^® presented the best performance, although with aheterogeneous accuracy, depending on MIC values.     

EVALUATION OF THE MOLLUSCICIDAL POTENTIAL OF HYDROALCOHOLIC EXTRACTS OF Jatropha gossypiifolia Linnaeus, 1753 ON Biomphalaria glabrata (Say, 1818)

The action of extracts from the stem, leaves, and fruit of Jatropha gossypiifolia on Biomphalaria glabrata was studied by analyzing survival, feeding capacity and oviposition ability. The extracts were obtained by macerating the plant parts in 92% ethanol, which were then evaporated until a dry residue was obtained and phytochemically studied. The molluscicidal activity on B. glabrata was investigated using the procedures recommended by WHO (1965). The amount of food ingested and oviposition were measured during each experiment. The extract of leaves from J. gossypiifolia was shown to be a strong molluscicidal agent, causing 100% mortality of B. glabrata, even in the lowest concentration tested, of 25 ppm. Regarding the fruit extract, there was variation in the mortality, depending on the concentration used (100, 75, 50 and 25 ppm). The snails that were in contact with the fruit extract had significant reduction in feeding and number of embryos in comparison to the control. The stem extract did not present molluscicidal activity nor had any influence on the feeding and oviposition abilities of B. glabrata, in the concentrations tested. In conclusion, the extracts of leaves and fruits of J. gossypiifolia investigated in this work show molluscicidal effect and may be sources of useful compounds for the schistosomiasis control.