Abstract in English:In recent years, the polymerase chain reaction (PCR) technique has significantly advanced towards expanding its use and versatility by working with quantitative real-time PCR (qPCR). Data from the literature show that both methods present interesting characteristics for the diagnosis of visceral leishmaniasis. The benefits of qPCR in relation to conventional PCR include speed, reproducibility and quantitative ability. In addition to operational advantages, qPCR is more sensitive and reproducible and may replace conventional PCR in diagnostic routines. Regarding visceral leishmaniasis, the possibility of deployment of real-time PCR in highly complex diagnoses (reference services) in endemic areas will facilitate a swift and safe return for patients. Moreover, the use of a technique that possesses elevated diagnostic sensitivity, and can monitor therapy and prevent relapses promotes broader prospects for the disease control.
Abstract in English:A comprehensive and up to date summary of the literature on the helminth parasites of lizards and amphisbaenians from South America is herein presented. One-hundred eighteen lizard species from twelve countries were reported in the literature harboring a total of 155 helminth species, being none acanthocephalans, 15 cestodes, 20 trematodes and 111 nematodes. Of these, one record was from Chile and French Guiana, three from Colombia, three from Uruguay, eight from Bolivia, nine from Surinam, 13 from Paraguay, 12 from Venezuela, 27 from Ecuador, 17 from Argentina, 39 from Peru and 103 from Brazil. The present list provides host, geographical distribution (with the respective biome, when possible), site of infection and references from the parasites. A systematic parasite-host list is also provided.
Abstract in English:Clostridium perfringens is an important pathogen that provokes numerous different diseases. This bacterium is classified into five different types, each of which capable of causing a different disease. There are various methods for the bacterial identification, many are labor-intensive, time-consuming, expensive and also present low sensitivity and specificity. The aim of this research was to identify the different types of C. perfringens using PCR molecular method. In this study, 130 sheep-dung samples were randomly collected from areas around the city of Kerman, southeastern Iran. After processing and culturing of samples, the produced colonies were morphologically studied, gram stain test was also carried out and the genera of these bacteria were identified through biochemical tests. DNA extracted from isolated bacteria for genotyping was tested by multiplex PCR with specific primers. Based on length of synthesized fragments by PCR, toxin types and bacterial strains were detected. C. perfringens isolated types were divided as follows: 17.39% type A, 21.74% type B, 34.78% type C and 26.09% type D. It should be emphasized that, up to the present moment, C. perfringens type A has not been reported in Iran.
Abstract in English:The venom of poisonous snakes comprises a complex mixture of several proteins with other less significant constituents, resulting in principles capable of changing viable tissues. The hemotoxic factor is the main responsible for necrosis and tissue sloughing. Envenomations are common in rural areas of Turkey caused by snake species that present hepatotoxic venom, which causes local swelling, ecchymosis and alterations in blood profile. The epidemiological and clinical findings of snake envenomations in Turkey were evaluated based on data recorded by the National Poison Information Center (NPIC) between 1995 and 2004, in a total of 550 snakebite cases. The month of peak incidence was June (24.3%) while most incidents occurred in Marmara, Central Anatolia and Black Sea regions of Turkey. The victims were mainly adults (54.1%). Hospitalized patients displayed clinical signs of local (75.2%) and systemic effects (24.7%). Local clinical symptoms comprised edema, pain, hyperemia, numbness and ecchymosis, while systemic clinical symptoms included nausea, vomiting, hypotension, tachycardia, dyspnea, dry mouth, paresthesia, generalized edema, cyanosis and compartment syndrome. Occasionally, convulsions, confusion, loss of consciousness, hyperthermia, hepatic and circulation failure, hematoma, drowsiness, epistaxis, chest and abdominal pain, venous spasm, thrombocytopenia and bradycardia were recorded. Approximately one third (34.2%) of the patients were treated symptomatically, while 10.5% required antivenom therapy along with symptomatic treatments and 26.3% of all patients were exclusively treated with antivenom. Although a significant number of incidents were reported, no deaths occurred. These findings emphasize the presence of multiple medically important snake species in Turkey and that public awareness and therapeutic approaches appear sufficient to manage snakebite incidents.
Abstract in English:A total of 310 blood smears were collected from sheep of the Livestock Experiment Station, Qadirabad, Sahiwal district, Pakistan, and surrounding areas. The samples were examined microscopically and 30 (9.67%) were positive for babesiosis. The animals were divided into two groups (A and B) for chemotherapy. Group A sheep were treated with diminazene diaceturate while group B animals received imidocarb dipropionate. Drug efficacy was determined by negative blood smear examination. Diminazene diaceturate effectiveness against babesiosis was 80% while that of imidocarb dipropionate was 100%. Hematological studies revealed a significant decrease in hemoglobin concentrations and hematocrit values for Babesia-positive animals compared to healthy controls.
Abstract in English:Interactions among microorganisms may be the cause of morphological modifications, particularly in fungal cells. The aim of this work was to examine the changes that occur in cells of the fungus Fonsecaea pedrosoi after in vitro co-culturing with Bacillus subtilis and to explore the results of this interaction in vivo in an experimental murine infection. B. subtilis strain was inoculated into a 15-day pure culture of F. pedrosoi. In vitro, after 48 hours of co-culturing, the fungal cells were roundish. The secretion of fungal dark pigments and production of terminal chlamydoconidia were observed in hyphae after one week. In the in vivo study, two animal groups of 30 BALB/c mice each were employed. One group was inoculated intraperitoneally with hyphal fragments from the co-culture of bacteria and fungi; the other group was infected only with F. pedrosoi hyphae. After seven days of infection, both animal groups developed neutrophilic abscesses. Phagocytosis of bacilli by macrophages occurred at three days. At later periods, generally after 25 days, only roundish cells similar to sclerotic bodies remained in the tissues while hyphae were eliminated by 15 to 20 days. These fungal forms originated mainly from terminal chlamydoconidia. The co-culturing between bacteria and fungi may constitute a mechanism to rapidly obtain resistant fungal forms for host defenses, especially for chromoblastomycosis (CBM) experimental infections.
Abstract in English:Mitochondrial DNA (mtDNA) has been widely employed in phylogeographic and phylogenetic studies. In the present study, the genetic identification of the scorpion Androctonus crassicauda (Olivier, 1807) was carried out by using the 16S mitochondrial gene, since this scorpion represents the most important species in Turkey regarding scorpionism and antivenom production. Two genetic groups were found according to the sequence analysis results, while five different loci at the nucleotide level presented genetic variation in the 16S region when compared to a known A. crassicauda sequence data (GenBank, AJ277598). Nucleotide variations found in the current work constitute the first descriptive report for A. crassicauda. Moreover, future studies may enlighten the genetic and venom composition variations for this scorpion species.
Abstract in English:Road-killed wild animals have been for years used for surveillance of vectors of zoonotic pathogens and may offer new opportunities for eco-epidemiological studies. In the current study, fungal infection was evaluated by PCR and nested-PCR in tissue samples collected from 19 road-killed wild animals. The necropsies were carried out and samples were collected for DNA extraction. Results, using PCR with a panfungal primer and nested PCR with specific primers, indicated that some animals are naturally infected with Amauroascus aureus, Metarhizium anisopliae, Aspergillus flavus, Aspergillus oryzae, Emmonsia parva, Paracoccidioides brasiliensis or Pichia stipitis. The approach employed herein proved useful for detecting the environmental occurrence of several fungi, as well as determining natural reservoirs in wild animals and facilitating the understanding of host-pathogen relationships.
Abstract in English:Some snake venom proteins present enzymatic activities, such as L-amino acid oxidase (LAAO). The aim of this paper was to investigate the effect of Bothrops leucurus total venom (BleuTV) and its fraction LAAO (BleuLAAO) on bacteria, yeast, and promastigote forms of Leishmania amazonensis and Leishmania chagasi, and epimastigote forms of Trypanosoma cruzi. BleuTV was isolated with a Protein Pack 5PW® (Waters Corporation, USA), and several fractions were obtained. BleuLAAO was purified to high molecular homogeneity, and its N-terminal amino acid sequence shared a high degree of amino acid conservation with other LAAOs. BleuTV inhibited Staphylococcus aureus growth in a dose-dependent manner, with a minimum inhibitory concentration (MIC) of 25 μg/mL, which corresponded to its minimum lethal concentration (MLC). BleuTV also inhibited the growth of promastigote forms of L. chagasi and L. amazonensis, with respective IC50 values of 1.94 μg/mL and 5.49 μg/mL. Furthermore, it repressed T. cruzi growth with an IC50 of 1.14 μg/mL. However, BleuLAAO did not inhibit the growth of the microorganisms studied and was not toxic to macrophages. BleuTV had low toxicity against macrophages at the concentrations studied. In conclusion, whole venom from Bothrops leucurus inhibited the growth of some microorganisms, including S. aureus, Leishmania sp., and T. cruzi.
Abstract in English:Snake venoms comprise a highly complex mixture of proteins, which requires for their characterization the use of versatile two-dimensional electrophoresis techniques. In the present study, venoms obtained from eight snakes (Ophiophagus hannah, Naja kaouthia, Naja sumatrana, Bungarus fasciatus, Trimeresurus sumatranus, Tropidolaemus wagleri, Enhydrina schistosa and Calloselasma rhodostoma) commonly found in Malaysia were separated based on two independent properties, isoelectric point (pI) and molecular weight (MW). Many differences in snake venoms at the inter-family, inter-subfamily, inter-genus and inter-species levels were revealed. Notably, proteins from individuals of the Viperidae family - Trimeresurus sumatranus, Tropidolaemus wagleri and Calloselasma rhodostoma - were found to be numerous and scattered by the two-dimensional gel electrophoresis (2DE) specifically in regions between 37 and 100 kDa compared to the Elapidae venom proteins. The latter were clustered at the basic and lower molecular mass region (less than 20 kDa). Trains of spots were commonly observed, indicating that these proteins may be derived from post-translational modifications. Ophiophagus hannah (Elapidae) revealed a great amount of protein spots in the higher molecular mass range when compared to Enhydrina schistosa, Naja kaouthia, Naja sumatrana and Bungarus fasciatus. Overall 2DE showed large differences in the venom profile of each species, which might be employed as an ancillary tool to the identification of venomous snake species.
Abstract in English:Naja naja snake venom presents abundant thermostable peptides. Many of them possess useful pharmacological activity that may be employed for drug development. For the proteomic characterization of such toxins, in the present study, Naja naja venom solution was heated up to 100°C for 10, 30, 60, 120, 180 and 300 minutes and protein fractions of non-heated and heated venom were analyzed by two-dimensional nano-liquid chromatography coupled online with tandem mass spectrometry. After heating for 300 minutes, a total of 32 peptides were still detected in the supernatant. The identified peptides belong to the following groups: cardiotoxins, neurotoxins and cytotoxins. It was found that thermostable peptides are able to preserve their analgesic activity after a long heating time in formalin test. Mice injected with 15 μg/g of 60-minute heated venom or with 25 μg/g of 300-minute heated venom revealed even a better analgesic activity than those treated with lidocaine.
Abstract in English:Iranian scorpions belong mainly to the Buthidae and Scorpionidae families, distributed into 16 genera and 25 species. In Iran, similar to other parts of the world, there are a few known species of scorpions responsible for severe envenoming; amongst which Mesobuthus eupeus is the most common. Its venom contains several toxin fractions that may affect the ion channel. In the present study purification, labeling and biological evaluation of M. eupeus venom are described. For separation, soluble venom was loaded on a chromatography column packed with Sephadex G-50 gel. Subsequently, the fractions were collected according to UV absorption at a wavelength of 280 nm. Toxic fraction (F3) was loaded on an anionic ion exchanger resin and then on a cationic resin. Finally, toxic subfractions F3.1.6 and F3.1.9 were labeled with 99mTc and injected into normal mice to distinguish excretion pathway. The venom toxic fraction was successfully obtained in its purified form. Radiolabeling of toxic fractions was performed at high specific activity with radiochemical purity of more than 97 and 95% respectively for F3.1.6 and F3.1.9. Biodistribution studies in normal mice with two toxic fractions usually show rapid clearance of the compounds from blood and tissue except for kidneys. Since tissue distribution studies are very important for clinical purpose, the present findings suggest that 99mTc labeling of venom is a useful tool for in vivo studies and comprises an excellent approach to monitoring the process of biodistribution and kinetics of toxins.
Abstract in English:The presence of bees (Apis mellifera L.) in urban areas has increased in recent years due to environmental disturbances caused by humans. Bee migration to cities may provoke serious accidents, since some people present allergic reactions to their venoms. In Rio Claro city, São Paulo state, Brazil, the number of calls to the fire brigade for removal of bee swarms, and the number admissions in local hospitals due to bee stings were investigated during 2002 and 2003, and a correlation between these data and the average temperature, rainfall and relative humidity was found. The study period was divided into three phases according to the number of times that the fire brigade was called to remove swarms (263 times): January to July 2002 - 51 calls (19.39%); August 2002 to July 2003 - 140 calls (53.23%); and August to December 2003 - 72 calls (27.38%). A significant correlation among the number of calls, the local temperature and rainfall was detected. The number of accidents was not associated with environmental variables. Based on the current results, public activities for prevention of bee attacks may be developed to avoid unwanted contact between humans and these insects, and/or provide the appropriate management of the colonies.
Abstract in English:The aim of the current study was to evaluate the infection rate by Brucella spp. in wild and in captive animals. Serum samples from 121 animals (94 free-ranging and 27 captive) of different mammal species were evaluated. Sera were submitted to rose Bengal test (RBT) for screening and serum agglutination tests (SAT) and 2-mercaptoethanol test (2-ME) for confirmatory results. Nine animals (five free-ranging and four captive) tested positive in RBT, but negative in the confirmatory tests. Several domestic animal diseases that have control programs are not focused on wild reservoirs, such as brucellosis in Brazil. The study of new reservoirs in wildlife is essential to prevent emerging diseases.
Abstract in English:In the period between June 2008 and August 2009, three cases of stings of Euscorpius scorpions indigenous to Italy were treated at two different emergency departments (ED) in hospitals of the Piedmont region, northwest Italy: Santa Croce e Carle General Hospital in Cuneo, and Santissima Annunziata Hospital in Savigliano. Scorpion stings in Italy are rare and not well documented in the literature; this situation may raise doubts among medical personnel as to how such lesions are best treated. Analysis of the incidents confirms that the venom of Euscorpius do not provokes systemic poisoning in humans and in these cases even dermatological reactions were not significant.