RESUMO
In the fall of 2021, a significant mortality event in free-ranging Southern Lapwing (Vanellus chilensis) occurred on a soccer field in southern Brazil. Approximately 130 adult southern lapwings died after showing weakness and flaccid paralysis, characterized by the inability to move or fly and drooped wings. Due to the large number of animals affected, there was concern that they had been criminally poisoned. The affected birds were found to have ingested maggots in fresh poultry litter incorporated into the grass surface. Postmortem examinations of four southern lapwings revealed no significant gross and histological findings. Polymerase Chain Reaction (PCR) for influenza A virus, flavivirus, and paramyxovirus was negative. Based on the epidemiological and clinical findings and the negative viral results, a presumptive diagnosis of botulism was made. This diagnosis was confirmed through mouse bioassay and seroneutralization, which detected botulinum toxin type C. Maggots loaded with botulinum neurotoxins were the probable vehicle for intoxication in the outbreak. Considering the impact of avian botulism on wild bird populations, our results may help prevent similar outbreaks in the future.
Assuntos
Doenças das Aves , Botulismo , Charadriiformes , Doenças dos Roedores , Camundongos , Animais , Botulismo/diagnóstico , Botulismo/epidemiologia , Botulismo/veterinária , Doenças das Aves/epidemiologia , Animais Selvagens , Aves , Larva , Surtos de Doenças/veterinária , Doenças dos Roedores/epidemiologiaRESUMO
Paralytic Shellfish Poisoning is a potentially fatal syndrome, resulting from the filter-feeding activities of marine molluscs accumulating harmful neurotoxins naturally occurring in microalgae. Outbreaks are well recognised throughout most regions of the world, but with the highest levels of toxicity to date recorded in mussels from Argentina. Whilst toxicity has been documented for selected outbreaks over the years, testing has been conducted using a mouse bioassay. Consequently there is a need to establish baseline data utilising modern chemical detection methods, which also facilitate the quantification of individual toxin analogues, giving useful data on toxin profiles as well as total sample toxicity. In this study, 151 shellfish samples harvested from the marine waters of Argentina between 1980 and 2012 were subjected to analysis by liquid chromatography with fluorescence detection, since Jan 2019 the European Union reference method for PSP determination. Total PST concentrations were found to vary enormously throughout the coastline of Argentina, with higher levels of toxins found in the central regions of Rio Negro and Chubut. Toxin profiles in terms of molar percentage of total concentrations were dominated by the gonyautoxins GTX1&4 and GTX2&3, followed by C1&2, STX and dcGTX2&3, with minor levels of other analogues previously not reported in the country. Profiles were found to vary significantly, with statistical clusters of profile types associated with a wide range of factors, including species, spatial and temporal differences, as well as likely source microalgae species and potential toxin transformation pathways. Overall application of the chemical detection method has confirmed both the significant risk to shellfish consumers in Argentina with periodic outbreaks of extremely high toxin levels and a large variability in toxin profiles relating in part to previously reported variabilities in microalgal toxin content. The study has demonstrated the potential for the method to systematically study the relationships between toxicity, toxin profile, source phytoplankton and other environmental factors.
Assuntos
Bivalves , Intoxicação por Frutos do Mar , Animais , Argentina , Toxinas Marinhas , Frutos do Mar/análiseRESUMO
Free-range chickens may ingest oocysts of T. gondii present in the environment and consequently harbor virulent strains of this parasite in different tissues, without any clinical signs. Isolation of T. gondii through bioassays on mice and cats from naturally infected chicken tissues has been described in several countries, demonstrating the importance of free-range chickens in the transmission of this parasite. The aim of this study was the genotypic characterization of T. gondii isolates obtained from naturally infected free-range chickens in a rural area of the state of Rio Grande do Sul, Brazil. Brain and heart tissue from 12 chickens seropositive for T. gondii were processed using peptic digestion technique for parasite isolation. From 12 samples subjected to mouse bioassay, nine isolates were obtained. RFLP-PCR genotypic characterization was performed using 11 genetic markers: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genetic characterization of the isolates revealed the presence of five atypical genotypes according to ToxoDB (# 11, # 55, # 64, # 140 and # 163). Our results showed a wide genetic diversity of T. gondii in free-range chickens in this region.(AU)
Galinhas criadas ao ar livre podem ingerir oocistos de T. gondii presentes no ambiente e, com isso, albergar cepas virulentas desse parasita em diferentes tecidos, sem sinais clínicos. O isolamento de T. gondii por meio de bioensaios em camundongos e gatos, a partir de tecidos de galinhas naturalmente infectadas, tem sido descrito em vários países. Isso demonstra a importância das galinhas caipiras na epidemiologia desse parasita. O objetivo deste trabalho foi caracterizar genotipicamente isolados de T. gondii obtidos de galinhas caipiras naturalmente infectadas em uma área rural do município de Santa Maria, estado do Rio Grande do Sul, Brasil. Fragmentos de cérebro e de coração, de 12 galinhas soropositivas para T. gondii, foram processados pela técnica de digestão péptica para isolamento do parasita. Das 12 amostras submetidas a bioensaio com camundongos, nove isolados foram obtidos. A caracterização genotípica por RFLP-PCR foi realizada utilizando-se 11 marcadores genéticos: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 e Apico e revelou a presença de cinco genótipos atípicos de acordo com o ToxoDB (# 11, # 55, # 64, # 140 e # 163). Os resultados mostraram uma ampla diversidade genética de T. gondii em galinhas caipiras nessa região.(AU)
Assuntos
Animais , Camundongos , Toxoplasma , Bioensaio/veterinária , Galinhas/virologia , Toxoplasmose Animal , Técnicas de Genotipagem/veterinária , Zona Rural , Reação em Cadeia da Polimerase/veterináriaRESUMO
Free-range chickens may ingest oocysts of T. gondii present in the environment and consequently harbor virulent strains of this parasite in different tissues, without any clinical signs. Isolation of T. gondii through bioassays on mice and cats from naturally infected chicken tissues has been described in several countries, demonstrating the importance of free-range chickens in the transmission of this parasite. The aim of this study was the genotypic characterization of T. gondii isolates obtained from naturally infected free-range chickens in a rural area of the state of Rio Grande do Sul, Brazil. Brain and heart tissue from 12 chickens seropositive for T. gondii were processed using peptic digestion technique for parasite isolation. From 12 samples subjected to mouse bioassay, nine isolates were obtained. RFLP-PCR genotypic characterization was performed using 11 genetic markers: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico. Genetic characterization of the isolates revealed the presence of five atypical genotypes according to ToxoDB (# 11, # 55, # 64, # 140 and # 163). Our results showed a wide genetic diversity of T. gondii in free-range chickens in this region.(AU)
Galinhas criadas ao ar livre podem ingerir oocistos de T. gondii presentes no ambiente e, com isso, albergar cepas virulentas desse parasita em diferentes tecidos, sem sinais clínicos. O isolamento de T. gondii por meio de bioensaios em camundongos e gatos, a partir de tecidos de galinhas naturalmente infectadas, tem sido descrito em vários países. Isso demonstra a importância das galinhas caipiras na epidemiologia desse parasita. O objetivo deste trabalho foi caracterizar genotipicamente isolados de T. gondii obtidos de galinhas caipiras naturalmente infectadas em uma área rural do município de Santa Maria, estado do Rio Grande do Sul, Brasil. Fragmentos de cérebro e de coração, de 12 galinhas soropositivas para T. gondii, foram processados pela técnica de digestão péptica para isolamento do parasita. Das 12 amostras submetidas a bioensaio com camundongos, nove isolados foram obtidos. A caracterização genotípica por RFLP-PCR foi realizada utilizando-se 11 marcadores genéticos: SAG1, 5'-3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 e Apico e revelou a presença de cinco genótipos atípicos de acordo com o ToxoDB (# 11, # 55, # 64, # 140 e # 163). Os resultados mostraram uma ampla diversidade genética de T. gondii em galinhas caipiras nessa região.(AU)
Assuntos
Animais , Camundongos , Toxoplasma , Bioensaio/veterinária , Galinhas/virologia , Toxoplasmose Animal , Técnicas de Genotipagem/veterinária , Zona Rural , Reação em Cadeia da Polimerase/veterináriaRESUMO
We report a case of severe congenital toxoplasmosis that involved an atypical T. gondii genotype in a newborn baby from Alagoas state in Northeastern Brazil. A pregnant woman presented IgM and IgG anti-T. gondii antibodies, as detected by the chemiluminescence immunoassay on the second trimester of pregnancy. A mouse bioassay was performed using umbilical cord blood and one isolate was obtained. The isolate was designated TgCTBrAL1 and genetic characterization revealed genotype ToxoDB #162. Genotype results of the rhoptry genes, ROP5 and ROP18, could predict the high virulence of the isolate in mice, which was confirmed by an in vivo virulence assay. This is the first report of generating a T. gondii isolate from a newborn baby with congenital toxoplasmosis in Northeastern Brazil.
Assuntos
Toxoplasma/isolamento & purificação , Toxoplasmose Congênita/parasitologia , Animais , Brasil , Genótipo , Humanos , Recém-Nascido , Masculino , Camundongos , Proteínas de Protozoários/genética , Toxoplasma/genética , Toxoplasma/patogenicidade , Virulência/genéticaRESUMO
The present study evaluated the viability of Toxoplasma gondii tissue cysts in dry-aged pork loins (m. longissimus) after 14, 21 and 28 days under controlled temperature (0⯰C⯱â¯1⯰C). The pigs (nâ¯=â¯9) were orally inoculated with 3,000 T. gondii oocysts. The right loin of each pig was aged for a predetermined period, and the left loin was kept unprocessed as a control. Two experiments were performed. In Experiment 1, the loins of three pigs were aged for 14 days and then bioassayed in both cats and mice. In Experiment 2, the loins of six pigs were bioassayed only in mice, and the ageing periods were 14, 21, and 28 days. Toxoplasma gondii tissue cysts remained viable in loins aged up to 14 days, as confirmed by bioassays in cats and mice. Viable T. gondii was not recovered by bioassays in mice from loins that were aged for 21 or 28 days. These results demonstrate that T. gondii remained viable in vacuum-packed dry-aged pork loins for 14 days at controlled temperature but not for 21 days or longer.
Assuntos
Carne/parasitologia , Doenças dos Suínos/parasitologia , Toxoplasma/crescimento & desenvolvimento , Toxoplasmose Animal/parasitologia , Animais , Bioensaio , Gatos , Embalagem de Alimentos/instrumentação , Camundongos , Oocistos/crescimento & desenvolvimento , Suínos , VácuoRESUMO
Toxoplasma gondii is a zoonotic parasite which can infect almost all warm-blooded animals. Toxoplasma gondii isolates from Brazil have greater genetic diversity with a predominance of virulent and atypical genotypes, compared with the Northern Hemisphere. Considering that previous studies have demonstrated a high seroprevalence of T. gondii antibodies in animals from Fernando de Noronha Island, the aim of this study was to isolate, genetically characterize, and determine mouse virulence of isolates of T. gondii from livestock from this Brazilian island. Two T. gondii isolates were obtained by mouse bioassay from brain from one sheep and one pig. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22- 8, c29-2, PK1, L358, and Apico) and an atypical genotype of T. gondii (ToxoDB #146) was identified for both isolates. Genotyping of four ROP loci indicated different alleles for ROP16 and mouse virulence analysis revealed different profiles (intermediate and low virulence). This is the first report of this genotype being described in a pig and a sheep.
Assuntos
Toxoplasma/genética , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/parasitologia , Animais , Brasil , Variação Genética , Genótipo , Ilhas , Camundongos , Proteínas de Protozoários/genética , Ovinos , Suínos , Toxoplasma/classificação , Virulência/genéticaRESUMO
The present study aimed to isolate and genotype strains of T. gondii from pigs slaughtered for human consumption in South Brazil. Blood and tissues (heart, diaphragm, liver, tongue, and masseter) from 400 animals were collected at two slaughterhouses. Sera were obtained, and antibodies against T. gondii were detected by both indirect fluorescence antibody test (IFAT) and modified agglutination test (MAT). The tissues of animals that tested positive in MAT, IFAT, or both (cut-off ≥ 64) were bioassayed. Twenty-six (6.5%) of the 400 animals were positive by serology. A total of 18 (69.2%) out of those 26 were positive in the mouse bioassay. The isolates were characterized by using 10 PCR-RFLP genetic markers. Fourteen isolates were fully genotyped, and four isolates were genotyped using nine of the 10 markers. All isolates belonged to ToxoDB PCR-RFLP genotype #206. The present study reports on genotype #206 in pigs for the first time, and it confirms the atypical nature of the Brazilian T. gondii isolates. Additionally, even with low levels of antibodies detected in pig herds, pork presents a T. gondii infection risk for humans.
Assuntos
Doenças Transmitidas por Alimentos/parasitologia , Doenças dos Suínos/parasitologia , Toxoplasma/genética , Toxoplasma/isolamento & purificação , Toxoplasmose Animal/parasitologia , Matadouros , Testes de Aglutinação/veterinária , Animais , Anticorpos Antiprotozoários/sangue , Bioensaio , Brasil , Técnica Indireta de Fluorescência para Anticorpo , Marcadores Genéticos , Variação Genética , Genótipo , Coração/parasitologia , Humanos , Fígado/parasitologia , Camundongos , Reação em Cadeia da Polimerase/veterinária , Polimorfismo de Fragmento de Restrição , Carne Vermelha/parasitologia , Suínos , Toxoplasmose Animal/sangueRESUMO
Botulinum neurotoxin type-A (BoNTA) is one of the seven different serotypes (A to G) produced by Clostridium botulinum. A stability-indicating size-exclusion chromatography (SEC) method was developed and validated, and the specificity was confirmed by forced degradation study, interference of the excipients, and peaks purity. The method was applied to assess the content and high-molecular-weight (HMW) forms of BoNTA in biopharmaceutical products, and the results were compared with those of the LD50 mouse bioassay, the T-47D cell culture assay, and the reversed-phase chromatography (RPC) method, giving mean values of 0.71% higher, 0.36% lower, and 0.87% higher, respectively. Aggregated forms showed significant effects on cytotoxicity, as well as a decrease in the bioactivity (p < 0.05). The employment of the proposed method in conjunction with the optimized analytical technologies for the analysis of the intact and altered forms of the biotechnology-derived medicines, in the correlation studies, enabled the demonstration of the capability of each one of the methods and allowed for great improvements, thereby assuring their safe and effective use.
Assuntos
Toxinas Botulínicas Tipo A/análise , Toxinas Botulínicas Tipo A/toxicidade , Animais , Bioensaio , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Cromatografia em Gel , Cromatografia Líquida , Cromatografia de Fase Reversa , Feminino , Humanos , Dose Letal Mediana , Camundongos , Reprodutibilidade dos TestesRESUMO
A large outbreak of botulism in feedlot steers fed corn silage contaminated with Clostridium botulinum neurotoxin type C (BoNT/C) is reported occurring in Midwestern Brazil in August 2017. The onset of the outbreak occurred 15 days after 1700 steers started to be fed the contaminated corn silage. Affected steers were alert and afebrile with varying degrees of flaccid paralysis in various muscle groups. A total of 1100 steers were affected, 1090 of which died within four days. Ten steers recovered after treatment with antitoxin. No gross or microscopic lesions were found in affected steers. The diagnosis was based on epidemiological data, characteristic clinical signs, and positive mouse bioassay results. This outbreak is interesting due to the high number of fatally affected cattle and the on-site diagnostic approach. This case report demonstrates the difficulties in diagnosing and treating botulism in cattle.
Assuntos
Botulismo/veterinária , Doenças dos Bovinos/epidemiologia , Clostridium botulinum tipo C/isolamento & purificação , Surtos de Doenças , Silagem/microbiologia , Zea mays/microbiologia , Animais , Botulismo/epidemiologia , Botulismo/patologia , Brasil , Bovinos , Doenças dos Bovinos/patologia , Silagem/efeitos adversos , Zea mays/efeitos adversosRESUMO
Toxoplasma gondii isolates from Brazil have a different phenotypic and genotypic pattern, with predominance of virulent isolates and recombinant genotypes, compared to the North Hemisphere. Considering that a new T. gondii genotype, non-pathogenic to mice, was previously identified from free-range chickens from the Fernando de Noronha Island, Brazil, this study aimed to identify genotypes of this parasite in tissue samples of feral cats (Felis catus) from this Brazilian Island. Anti-T. gondii IgG antibodies were detected in 18/31 (58%) feral cats. Two non-virulent T. gondii isolates were obtained by mouse bioassay. Genotyping was performed by PCR-RFLP using 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, L358 and Apico) and an atypical strain of T. gondii (ToxoDB #146) was identified. This is the first report of this genotype in feral cats.
Assuntos
Toxoplasma/genética , Toxoplasma/patogenicidade , Toxoplasmose Animal/parasitologia , Animais , Anticorpos Antiprotozoários/sangue , Brasil , Gatos , DNA de Protozoário/genética , Marcadores Genéticos/genética , Genótipo , Ilhas , Camundongos , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Toxoplasma/isolamento & purificação , Virulência/genéticaRESUMO
From 2010 to 2012, 35 ciguatera fish poisoning (CFP) events involving 87 individuals who consumed locally-caught fish were reported in Guadeloupe (French West Indies). For 12 of these events, the presence of ciguatoxins (CTXs) was indicated in meal remnants and in uncooked fish by the mouse bioassay (MBA). Caribbean ciguatoxins (C-CTXs) were confirmed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. Using a cell-based assay (CBA), and the only available standard Pacific ciguatoxin-1 (P-CTX-1), the lowest toxins level detected in fish samples causing CFP was 0.022 µg P-CTX-1 equivalent (eq.)·kg(-1) fish. Epidemiological and consumption data were compiled for most of the individuals afflicted, and complete data for establishing the lowest observable adverse effects level (LOAEL) were obtained from 8 CFP events involving 21 individuals. Based on toxin intakes, the LOAEL was estimated at 4.2 ng P-CTX-1 eq./individual corresponding to 48. 4 pg P-CTX-1 eq.kg(-1) body weight (bw). Although based on limited data, these results are consistent with the conclusions of the European Food Safety Authority (EFSA) opinion which indicates that a level of 0.01 µg P-CTX-1 eq.kg(-1) fish, regardless of source, should not exert effects in sensitive individuals when consuming a single meal. The calculated LOAEL is also consistent with the U.S. Food and Drug Administration guidance levels for CTXs (0.1 µg C-CTX-1 eq.kg(-1) and 0.01 µg P-CTX-1 eq.kg(-1) fish).
Assuntos
Ciguatera/induzido quimicamente , Ciguatoxinas/análise , Ciguatoxinas/toxicidade , Peixes/metabolismo , Alimentos Marinhos/análise , Alimentos Marinhos/toxicidade , Animais , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Ciguatera/epidemiologia , Relação Dose-Resposta a Droga , Guadalupe , Humanos , Masculino , Camundongos , Testes de Toxicidade/métodosRESUMO
The presence of phytoplankton responsible for the production of lipophilic marine biotoxins is well recognised throughout parts of South America. To date, the quantitation of lipophilic toxins in Argentinean shellfish has been limited to select and highly focussed geographical studies. This work reports the analysis for lipophilic marine biotoxins in shellfish harvested across five regions of Argentina between 1992 and 2012. LC-MS/MS analysis was used for the quantitation of all regulated lipophilic toxins. High concentrations of okadaic acid group toxins were quantified, with a clear dominance of the parent okadaic acid and more than 90% of the toxin present as esters. Results showed DSP toxins in shellfish from the Buenos Aires Province during 2006 and 2007, earlier than previously described. There was also strong evidence linking the presence of okadaic acid to human intoxications. Other lipophilic toxins detected were yessotoxin, pectenotoxin-2 and 13-desMeC spirolide. With evidence published recently for the presence of azaspiracid producers, this work reports the detection of low concentrations of azaspiracid-2 in shellfish. As such the data provides the first published evidence for yessotoxins and azaspiracids in Argentinean shellfish and further evidence for the continuing presence of lipophilic marine toxins in Argentinean waters.
Assuntos
Contaminação de Alimentos/análise , Toxinas Marinhas/análise , Ácido Okadáico/análise , Fitoplâncton/química , Frutos do Mar/análise , Animais , Argentina , Cromatografia Líquida , Furanos/análise , Humanos , Macrolídeos , Venenos de Moluscos , Oxocinas/análise , Piranos/análise , Intoxicação por Frutos do Mar , Compostos de Espiro/análise , Espectrometria de Massas em TandemRESUMO
Here, we report different lipophilic toxins (LTs) detected by LC-MS/MS in Mediterranean mussels (Mytilus galloprovincialis) collected through 2012 in Todos Santos Bay, northwest Baja California, Mexico. The concentration of okadaic acid (OA), dinophysistoxin 2 (DTX2), and pectenotoxin 2 (PTX2) reached 500 µg kg(-1) during July and increased to 1647 µg kg(-1) in October. These toxins were associated with the presence of Dinophysis fortii and Dinophysis acuminata and a strong stratification of the water column. Other LTs present were yessotoxins, with a maximum concentration of 1080 µg kg(-1) in June. Cyclic imines (13-desmethyl spirolide and gymnodimine) and azaspiracid 1 were also detected in the mussels but at low concentrations. Diarrhetic toxins concentrations evaluated by LC-MS/MS were compared with the results of two mouse bioassay protocols. Positive results were obtained with both MBA protocols in several samples that presented toxicities below 160 µg OA-eq kg(-1), as estimated by LC-MS/MS results whereas other samples returned negative MBA results in samples with concentrations above this level. Therefore, analytical methods need to be applied to confirm the presence of regulated LTs. This is the first report of LTs in mussels cultivated in Mexico. The occurrence of these toxins represents an emerging problem in the region.
Assuntos
Lipídeos/química , Mytilus/química , Ácido Okadáico/análise , Piranos/análise , Animais , Cromatografia Líquida , Macrolídeos , Toxinas Marinhas/análise , Toxinas Marinhas/toxicidade , Camundongos , Camundongos Endogâmicos ICR , Ácido Okadáico/toxicidade , Piranos/toxicidade , Espectrometria de Massas em TandemRESUMO
Cnidarians comprise an old and diverse animal phylum, and possess a wide variety of biologically active substances. Sea anemones contain a diversity of interesting biologically active compounds including some potent toxins. In the present work, the sea anemones Stichodactyla mertensii and Stichodactyla gigantea, collected from the Mandapam coast, are characterized biomedically and pharmacologically. The crude protein was obtained by using methanol and aqueous extracts. The respective protein contents of S. mertensii and S. gigantea were found to be 2.10 µg/mL and 1.87 µg/mL. The methanol and aqueous extracts of S. mertensii and S. gigantea yielded six and nine bands by SDS-PAGE on 12 percent gel. In the hemolytic assay, both extracts exhibited hemolytic effect on chicken, goat, cow and human erythrocytes ('A', 'B' and 'O'). The neurotoxic effects of these crude extracts were determined in vivo using the sea shore crab Ocypode macrocera and mortality was observed. The mouse bioassay for lethality was performed on male albino mice. The crude extract of S. mertensii showed higher lethality (58 seconds at 1 mL-dose) than that of S. gigantea (2 minutes and 10 seconds at 0.75 mL-dose). The analgesic activity test was also carried out on albino mice by Eddy's hot plate and tail-flick methods. The extracts showed moderate analgesic effect by both hot-plate and tail-flick methods. These characteristics emphasize the need for the isolation and molecular characterization of new active toxins in S. mertensii and S. gigantea.(AU)
Assuntos
Animais , Venenos de Cnidários/administração & dosagem , Venenos de Cnidários/farmacologia , Fauna Aquática/análise , Metanol/análise , MortalidadeRESUMO
Cnidarians comprise an old and diverse animal phylum, and possess a wide variety of biologically active substances. Sea anemones contain a diversity of interesting biologically active compounds including some potent toxins. In the present work, the sea anemones Stichodactyla mertensii and Stichodactyla gigantea, collected from the Mandapam coast, are characterized biomedically and pharmacologically. The crude protein was obtained by using methanol and aqueous extracts. The respective protein contents of S. mertensii and S. gigantea were found to be 2.10 µg/mL and 1.87 µg/mL. The methanol and aqueous extracts of S. mertensii and S. gigantea yielded six and nine bands by SDS-PAGE on 12 percent gel. In the hemolytic assay, both extracts exhibited hemolytic effect on chicken, goat, cow and human erythrocytes ('A', 'B' and 'O'). The neurotoxic effects of these crude extracts were determined in vivo using the sea shore crab Ocypode macrocera and mortality was observed. The mouse bioassay for lethality was performed on male albino mice. The crude extract of S. mertensii showed higher lethality (58 seconds at 1 mL-dose) than that of S. gigantea (2 minutes and 10 seconds at 0.75 mL-dose). The analgesic activity test was also carried out on albino mice by Eddy's hot plate and tail-flick methods. The extracts showed moderate analgesic effect by both hot-plate and tail-flick methods. These characteristics emphasize the need for the isolation and molecular characterization of new active toxins in S. mertensii and S. gigantea.(AU)
Assuntos
Animais , Masculino , Ratos , Anêmonas-do-Mar/química , Antivenenos , Venenos de Cnidários/toxicidade , Neurotoxinas/química , Bioensaio/métodos , Proteínas Hemolisinas/isolamento & purificação , Proteínas Hemolisinas/efeitos dos fármacos , Analgésicos/farmacologiaRESUMO
The distribution of T. gondii in commercial cuts of pork (ham, tenderloin, spareribs and arm picnic) by PCR and bioassay from experimentally infected pigs, was evaluated. Eighteen mixed breed pigs were divided into two groups (G). The G1 animals (n=10) were infected with 4 x104 oocysts of the T. gondii VEG strain and the G2 animals (n=8) were used as control. Pigs of both groups were slaughtered at 59th day after infection, and meat samples were collected for bioassay and PCR. All animals from G1 were positive by at least one or both tests, and all control animals were negative. T. gondii was identified in pork by mouse bioassay and PCR in 27/40 (67.5%) and in 9/40 (22.5%) of the evaluated samples, respectively. There were no statistical differences in the distribution of tissue cysts from commercial cuts of pork by bioassay (P>0.05). However, statistical differences were observed when mouse bioassay and PCR were compared (P<0.01).(AU)
Avaliou-se a presença de T. gondii em cortes comerciais de carne suína (pernil, lombo, costela e paleta), por meio do bioensaio e PCR, em animais experimentalmente inoculados. Dois grupos (G) foram formados. Os animais do G1 (n=10) foram inoculados com 4 x104 oocistos da cepa VEG e os do G2 (n=8) permaneceram como grupo-controle, não inoculado. Todos os animais foram abatidos no dia 59 após a infecção, quando foram colhidas as amostras de carne para a realização das provas de bioensaio e da PCR. Todos os suínos do G1 apresentaram-se positivos a pelo menos um dos testes de diagnóstico ou a ambos, e os do grupo-controle permaneceram negativos. Não houve diferenças significativas em relação aos tipos de cortes comerciais e à presença do parasita no bioensaio (P>0,05). O bioensaio foi capaz de detectar T. gondii em 27/40 (67,5%) amostras e a PCR em 9/40 (22,5%). O estudo mostrou diferença entre o bioensaio e a PCR (P<0,01).(AU)