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1.
Rev Soc Bras Med Trop ; 57: e00806, 2024.
Article in English | MEDLINE | ID: mdl-39082524

ABSTRACT

This report describes the occurrence of the rabies virus in two species of wild animals in the urban area of Montes Claros (MOC), Minas Gerais State, Brazil, in May 2023. The virus has been detected in frugivorous chiropterans (Artibeus sp) and marmosets (Callithrix penicillata). This is the first notified case of the rabies virus in the species C. penicillata in the urban area of MOC. Our findings show that the rabies virus is circulating in the urban area of MOC; therefore, permanent preventive measures must be adopted to avoid infection of other animals and humans.


Subject(s)
Callithrix , Rabies virus , Rabies , Animals , Rabies virus/isolation & purification , Brazil/epidemiology , Callithrix/virology , Rabies/veterinary , Rabies/epidemiology , Chiroptera/virology , Animals, Wild/virology
2.
Anaerobe ; 85: 102817, 2024 Feb.
Article in English | MEDLINE | ID: mdl-38163631

ABSTRACT

OBJECTIVES: This study aimed to produce and purify Clostridium perfringens type C beta-toxin, sheep anti-beta toxin immunoglobulin G (IgG) and chicken immunoglobulin Y (IgY). METHODS: Two methods were used for beta-toxin purification: single-step metal affinity chromatography (MAC) using zinc as a chelator and ion exchange chromatography (IEX). The purified and inactivated beta-toxoids were then administered to sheep and chickens in order to produce IgG and IgY. RESULTS: All assays using the IEX failed. In contrast, MAC purified more than 21 mg of toxin per run in a single-step protocol. The purified and inactivated beta-toxoids were then administered to sheep and chickens, and IgG and IgY were purified with a high yield, medium antibody titer of 50 IU/mL, and high avidity (73.2 %). CONCLUSIONS: C. perfringens type C beta-toxin and sheep or chicken anti-beta toxin IgG and IgY antibodies were successfully produced and purified using a simple protocol. This protocol can be used for the production of components used in the diagnosis and research of necrotic enteritis caused by C. perfringens type C, as well as for the evaluation of existing vaccines and the development of new preventive methods against this disease.


Subject(s)
Antitoxins , Clostridium Infections , Enteritis , Immunoglobulins , Poultry Diseases , Animals , Sheep , Clostridium perfringens , Clostridium Infections/veterinary , Enteritis/veterinary , Chickens , Toxoids , Immunoglobulin G , Poultry Diseases/prevention & control
3.
Rev. Soc. Bras. Med. Trop ; Rev. Soc. Bras. Med. Trop;57: e00806, 2024. graf
Article in English | LILACS-Express | LILACS | ID: biblio-1569578

ABSTRACT

ABSTRACT This report describes the occurrence of the rabies virus in two species of wild animals in the urban area of Montes Claros (MOC), Minas Gerais State, Brazil, in May 2023. The virus has been detected in frugivorous chiropterans (Artibeus sp) and marmosets (Callithrix penicillata). This is the first notified case of the rabies virus in the species C. penicillata in the urban area of MOC. Our findings show that the rabies virus is circulating in the urban area of MOC; therefore, permanent preventive measures must be adopted to avoid infection of other animals and humans.

4.
Anaerobe ; 72: 102465, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34662696

ABSTRACT

Herd vaccination is an important preventive measure against enterotoxemia in ruminants. Vaccination in goats should be performed every four months, and recent studies have shown that immunity in cattle lasts for less than one year. One of the mechanisms for increasing the duration of the immune response is to use purified toxoids as immunogens. The aim of the present study was to evaluate the humoral response in cattle and goats after vaccination with purified and semi-purified Clostridium perfringens type D epsilon toxoid. The following three different vaccines were used: vaccine 1 (V1), a semi-purified toxoid adsorbed to aluminum hydroxide; vaccine 2 (V2), a purified toxoid adsorbed to aluminum hydroxide; and vaccine (V3), a purified toxoid adsorbed on chitosan microparticles. Groups of cattle (n = 6-7) and goats (n = 6-7) were vaccinated on days 0 and 30, and serum samples for antitoxin titration were collected every 30 days for one-year post-vaccination. Goats were revaccinated on day 360, and their serum was evaluated on days 367 and 374. The antibody peaks ranged between 6.90 and 11.47 IU/mL in cattle and from 1.11 to 4.40 IU/mL in goats. In cattle administered with the V1 and V2 vaccines, we observed that the antibody titers were maintained above 0.2 IU/mL until the end of the experiment. In goats, V2 elicited long-lasting antibodies, and all animals maintained the protective titers for 210 days after the first dose. In conclusion, the purified toxoid vaccine with aluminum hydroxide adjuvant was able to induce strong and long-lasting humoral responses in both species and could be an alternative for improving the immunization schedule against enterotoxemia in goats and cattle.


Subject(s)
Bacterial Toxins/immunology , Cattle Diseases/immunology , Cattle Diseases/microbiology , Clostridium Infections/veterinary , Clostridium perfringens/immunology , Goat Diseases/microbiology , Goat Diseases/prevention & control , Toxoids/administration & dosage , Animals , Antibodies, Bacterial/blood , Antibodies, Bacterial/immunology , Bacterial Toxins/administration & dosage , Bacterial Toxins/chemistry , Bacterial Vaccines/administration & dosage , Bacterial Vaccines/chemistry , Bacterial Vaccines/immunology , Cattle , Clostridium perfringens/classification , Enterotoxemia/prevention & control , Goats , Immunity, Humoral , Immunization , Rabbits
5.
J Anim Sci ; 99(5)2021 May 01.
Article in English | MEDLINE | ID: mdl-33822982

ABSTRACT

The aim of this study was to identify possible effects of different vaccination strategies (concomitantly or not) against brucellosis and clostridia on intake, performance, feeding behavior, blood parameters, and immune responses of dairy heifers calves. Fifty heifers calves were enrolled [38 Gyr (Zebu, Bos taurus indicus) and 12 5/8 Holstein × Gyr]. At 120 d of age, animals were randomly distributed among 3 groups: B (n = 18), vaccinated against brucellosis; C (n = 14), vaccinated against clostridia and CB (n = 18), vaccinated concomitantly for both. Rectal and thermographic temperatures were evaluated on days -1, 0, 1, 2, 3, 5, 7,10, 14, and 28 relatives to the vaccination day. Feed and water intake, body weight (BW), and feeding behavior were monitored daily by an electronic feeding system. Blood was sampled on days 0, 3, 7, 14, and 28, relative to the vaccination day for determination of glucose and ß -hydroxybutyrate (BHBA) concentrations. Blood sampled on day 0 (prevaccination) and on days 28 and 42 were used to evaluate the immune response against Brucella abortus and clostridia. There was an increase in rectal temperature between the first and the third day postvaccination in the 3 groups. The thermography revealed an increase of local temperature for 7 d on groups B and CB. Group C had increased local temperature for a longer period, lasting for up to 14 d. Dry mater intake was reduced for groups B and CB, but no alteration was observed for group C. No alterations regarding initial BW, final BW, average daily weight gain, and feed efficiency were observed. No differences were observed for the 3 vaccination groups for blood parameters throughout the evaluation period. The concomitant vaccination against brucellosis and clostridia led to lower neutralizing antibody titers against epsilon toxin of Clostridium perfringens and botulinum toxin type C of C. botulinum (C > CB > B). When cellular proliferation assay and serological tests to B. abortus were evaluated, no differences were observed between groups B and CB. The present results indicate that the concomitant vaccination against brucellosis and clostridia has no relevant impact on the intake, performance, and feeding behavior of dairy calves. However, the concomitant vaccination of vaccines against these 2 pathogens impacts animal immunity against clostridial infections.


Subject(s)
Brucellosis , Cattle Diseases , Animal Feed/analysis , Animals , Body Weight , Brucellosis/veterinary , Cattle , Cattle Diseases/prevention & control , Feeding Behavior , Female , Immunity , Vaccination/veterinary
6.
Anaerobe ; 69: 102354, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33675994

ABSTRACT

The aim of this study was to purify Clostridium perfringens type D epsilon toxin and produce and purify anti-epsilon chicken immunoglobulin Y (IgY). A single-step ion exchange chromatography resulted in a high-yield and high-purity toxin, while ion exchange chromatography followed by gel filtration resulted in the highest purity of the toxin, but at a lower yield. Purified and inactivated epsilon toxin were then administered in chickens via four inoculations and IgY was obtained at a high purity and yield, with an antibody titer of 50 IU/mL and high levels of avidity (73.2%). In summary, C. perfringens type D epsilon toxin and chicken anti-epsilon IgY were successfully produced and purified, and may be used for the diagnosis of enterotoxemia caused by the epsilon toxin, as well as in potency tests of existing and future vaccines against enterotoxemia.


Subject(s)
Antibodies, Bacterial/blood , Bacterial Toxins/biosynthesis , Bacterial Toxins/isolation & purification , Chickens/microbiology , Clostridium perfringens/pathogenicity , Enterotoxemia/immunology , Enterotoxemia/physiopathology , Immunoglobulins/blood , Animals
7.
Anaerobe ; 59: 72-75, 2019 Oct.
Article in English | MEDLINE | ID: mdl-31129336

ABSTRACT

The aim of this study was to evaluate the titers of neutralizing antibodies in cattle inoculated with multivalent commercial clostridial vaccines containing C. botulinum type C (BoNTC), C. botulinum type D (BoNTD), and C. perfringens epsilon (ETX) toxoids for a period of one year. Cattle (Bos taurus), aged 4-6 months and not previously immunized, were vaccinated under four different protocols at days 0 and 30 and followed over one year. Individual serum titration was performed by a serum neutralization test in mice or in MDCK cells. The number of animals with detectable neutralizing antibodies ranged from 40.6% to 78.1%, but only 12.5% of animals showed neutralizing antibodies against all tested antigens. Neutralizing antibodies were found only until 60 days for ETX, 120 days for BoNTC, and 180 days for BoNTD. The absence of detectable neutralizing antibodies against the three antigens before 360 days, suggests that cattle remained unprotected for a long period before the recommended booster vaccination.


Subject(s)
Bacterial Toxins/immunology , Botulinum Toxins/immunology , Immunity, Humoral , Toxoids/immunology , Animals , Antitoxins/blood , Cattle , Dogs , Madin Darby Canine Kidney Cells , Mice , Neutralization Tests , Time Factors , Toxoids/administration & dosage
8.
Anaerobe ; 51: 47-49, 2018 Jun.
Article in English | MEDLINE | ID: mdl-29621603

ABSTRACT

Botulism is a well-known intoxication that affects humans and animals. The disease is endemic in cattle in Brazil and recently emerged as an important disease in commercial laying hens and broiler chickens in Europe. Dogs and other animal species can also be affected. Although antitoxins are commonly administered to humans diagnosed with botulism, in animals this is rarely the case and the treatment of botulism is still based only on support therapy. In the present work, we report an outbreak of type C botulism in Brazil that simultaneously affected domestic chickens, dogs and a black-pencilled marmoset (Callithrix penicillata). The successful use of Clostridium botulinum types C and D antitoxin for the treatment of an affected dog is also described.


Subject(s)
Botulism/veterinary , Clostridium botulinum type C/isolation & purification , Disease Outbreaks , Dog Diseases/epidemiology , Poultry Diseases/epidemiology , Animals , Antitoxins/therapeutic use , Botulism/epidemiology , Botulism/therapy , Brazil/epidemiology , Callithrix , Chickens , Dog Diseases/microbiology , Dog Diseases/therapy , Dogs , Poultry Diseases/microbiology , Treatment Outcome
9.
Semina Ci. agr. ; 37(1): 257-262, jan.-fev. 2016. tab
Article in English | VETINDEX | ID: vti-23158

ABSTRACT

Clostridium perfringens is a microorganism commonly found in the microbiota of humans and animals and a potential cause of enteric, muscle or nervous diseases. The treatment of these diseases is based on antimicrobial therapy and it is extremely important to know the antimicrobial susceptibility profile of the strains present in the region. The aim of this study was to evaluate the antimicrobial susceptibility of C. perfringens isolated from domestic and wild animals in Brazil against seven different antimicrobials. Forty-one strains from the stool samples of cattle (n = 12), buffalo (n = 2), goat (n = 3), dogs (n = 12) and wild carnivores (n = 12) were examined. The minimum inhibitory concentration was determined by the agar dilution method using Brucella agar supplemented with 5% of sheep blood, 0.1% of vitamin K, 0.1% of hemin and concentrations ranging from 0,25 to 256,0 mg L-¹ of the following antibiotics: erythromycin, florfenicol, metronidazole, oxytetracycline, penicillin, tylosin, and vancomycin. All C. perfringens strains were susceptible to florfenicol, metronidazole, penicillin and vancomycin. Two strains (4.9%) were resistant to erythromycin and tylosin, while five (12.2%) were resistant to oxytetracycline, one of which (2.4%) from an ocelot.(AU)


Clostridium perfringens é um microrganismo comumente encontrado na microbiota de seres humanos e animais e potencial causador de enfermidades entéricas, musculares ou neurológicas. O tratamento das enfermidades é baseado em terapia antimicrobiana, sendo de extrema importância conhecer o perfil de susceptibilidade antimicrobiana das estirpes presentes na região em questão. O presente estudo teve como objetivo avaliar a susceptibilidade de estirpes de C. perfringens isolados de animais domésticos e carnívoros silvestres no Brasil frente a sete diferentes antimicrobianos. Foram utilizados 41 isolados originários de bezerros (n = 12), búfalos (n = 2), caprinos (n = 3), cães (n = 12) e espécies de carnívoros silvestres (n = 12). A concentração inibitória minima foi determinada pelo método de diluição seriada em agar, utilizando-se o agar Brucella suplementado com 5% de sangue ovino, 0,1% de vitamina K, 0,1% de hemina e concentrações variando de 0,25 a 256,0 mg L-¹ dos seguintes antimicrobianos: eritromicina, florfenicol, metronidazol, oxitetraciclina, penicilina, tilosina e vancomicina. Todas as estirpes de C. perfringens testadas foram sensíveis ao florfenicol, metronidazol, penicilina e vancomicina. Dois isolados (4,9 %) foram resistentes a eritromicina e a tilosina, enquanto que cinco (12,2 %) estirpes foram resistentes a oxitetraciclina, sendo uma delas (2,4 %) proveniente de uma jaguatirica.(AU)


Subject(s)
Animals , Clostridium Infections/veterinary , Clostridium perfringens/genetics , Anti-Infective Agents/analysis , Drug Resistance, Microbial
10.
Semina ciênc. agrar ; 37(1): 257-262, 2016. tab
Article in English | VETINDEX | ID: biblio-1500277

ABSTRACT

Clostridium perfringens is a microorganism commonly found in the microbiota of humans and animals and a potential cause of enteric, muscle or nervous diseases. The treatment of these diseases is based on antimicrobial therapy and it is extremely important to know the antimicrobial susceptibility profile of the strains present in the region. The aim of this study was to evaluate the antimicrobial susceptibility of C. perfringens isolated from domestic and wild animals in Brazil against seven different antimicrobials. Forty-one strains from the stool samples of cattle (n = 12), buffalo (n = 2), goat (n = 3), dogs (n = 12) and wild carnivores (n = 12) were examined. The minimum inhibitory concentration was determined by the agar dilution method using Brucella agar supplemented with 5% of sheep blood, 0.1% of vitamin K, 0.1% of hemin and concentrations ranging from 0,25 to 256,0 mg L-¹ of the following antibiotics: erythromycin, florfenicol, metronidazole, oxytetracycline, penicillin, tylosin, and vancomycin. All C. perfringens strains were susceptible to florfenicol, metronidazole, penicillin and vancomycin. Two strains (4.9%) were resistant to erythromycin and tylosin, while five (12.2%) were resistant to oxytetracycline, one of which (2.4%) from an ocelot.


Clostridium perfringens é um microrganismo comumente encontrado na microbiota de seres humanos e animais e potencial causador de enfermidades entéricas, musculares ou neurológicas. O tratamento das enfermidades é baseado em terapia antimicrobiana, sendo de extrema importância conhecer o perfil de susceptibilidade antimicrobiana das estirpes presentes na região em questão. O presente estudo teve como objetivo avaliar a susceptibilidade de estirpes de C. perfringens isolados de animais domésticos e carnívoros silvestres no Brasil frente a sete diferentes antimicrobianos. Foram utilizados 41 isolados originários de bezerros (n = 12), búfalos (n = 2), caprinos (n = 3), cães (n = 12) e espécies de carnívoros silvestres (n = 12). A concentração inibitória minima foi determinada pelo método de diluição seriada em agar, utilizando-se o agar Brucella suplementado com 5% de sangue ovino, 0,1% de vitamina K, 0,1% de hemina e concentrações variando de 0,25 a 256,0 mg L-¹ dos seguintes antimicrobianos: eritromicina, florfenicol, metronidazol, oxitetraciclina, penicilina, tilosina e vancomicina. Todas as estirpes de C. perfringens testadas foram sensíveis ao florfenicol, metronidazol, penicilina e vancomicina. Dois isolados (4,9 %) foram resistentes a eritromicina e a tilosina, enquanto que cinco (12,2 %) estirpes foram resistentes a oxitetraciclina, sendo uma delas (2,4 %) proveniente de uma jaguatirica.


Subject(s)
Animals , Anti-Infective Agents/analysis , Clostridium perfringens/genetics , Clostridium Infections/veterinary , Drug Resistance, Microbial
11.
Clín. Vet. ; 19(108): 46-51, 2014.
Article in English, Spanish, Portuguese | VETINDEX | ID: vti-10361

ABSTRACT

O botulismo é uma intoxicação atribuída à ingestão de carne deteriorada ou crua, provocada por potentes toxinas de Clostridium botulinum, que causam tetraparesia flácida ascendente em cães. São sete tipos: A, B, C(?,?), D, E, F e G, com predomínio do C e D nos animais. Embora seja considerado raro na maioria dos países desenvolvidos, ele ainda pode ser encontrado com certa frequência. Este trabalho relata dez casos de botulismo canino tipo C, oriundos de área urbana e rural, atendidos entre os meses de maio de 2009 e outubro de 2010 em clínicas particulares, na cidade de Petrolina, Pernambuco. No presente trabalho, foram analisados a abordagem do histórico, os sinais clínicos, o diagnóstico laboratorial e o tratamento. Foi utilizada a técnica de inoculação em camundongo na identificação da toxina botulínica, que se mostrou sensível neste trabalho.(AU)


Botulism is a poisoning caused by ingestion of potent toxins of Clostridium botulinum, causing upward flaccid quadriplegia in dogs. There are seven types of botulism: A, B, C (?, ?), D, E, F and G, with the predominance of C and D in animals, attributed to the ingestion of raw or decaying meat. Although considered rare in most developed countries, it can still be found with some frequency. This article reports ten cases of canine type C botulism, originating from urban and rural areas and seen between the months of May 2009 and October 2010 in private veterinary practices in the city of Petrolina, Pernambuco, Brazil. The approach of anamnesis, clinical signs, and laboratory diagnosis were analyzed. The technique of inoculation in mice was used for the identification of the botulinum toxin, which has shown to be sensitive in this work.(AU)


El botulismo es una intoxicación que suele producirse por el consumo de carne en mal estado o cruda, provocada por la ingestión de potentes toxinas de Clostridium botulinum, que en los perros causan una tetraparesia fláxida ascendente. En los animales existen siete tipos de toxinas: A, B, C (?,?), D, E, F e G, con predominio de la C y D. A pesar de ser considerada una enfermedad de aparición rara en los países desarrollados, la misma puede ser encontrada con cierta frecuencia. Este trabajo relata diez casos de botulismo canino tipo C, originados en áreas urbanas y rurales, que fueron atendidos entre mayo de 2009 y octubre de 2010 en clínicas particulares de la ciudad de Petrolina, Pernambuco, Brasil. En este trabajo se analizaron el histórico del paciente, sus signos clínicos, exámenes de laboratorio y el tratamiento. Se utilizó una técnica de inoculación en ratones, que fue sensible para la identificación de la toxina botulínica.(AU)


Subject(s)
Animals , Dogs , Botulism , Poisoning/complications , Diagnosis , Dogs/classification
12.
Article in English, Spanish, Portuguese | VETINDEX | ID: biblio-1480952

ABSTRACT

O botulismo é uma intoxicação atribuída à ingestão de carne deteriorada ou crua, provocada por potentes toxinas de Clostridium botulinum, que causam tetraparesia flácida ascendente em cães. São sete tipos: A, B, C(?,?), D, E, F e G, com predomínio do C e D nos animais. Embora seja considerado raro na maioria dos países desenvolvidos, ele ainda pode ser encontrado com certa frequência. Este trabalho relata dez casos de botulismo canino tipo C, oriundos de área urbana e rural, atendidos entre os meses de maio de 2009 e outubro de 2010 em clínicas particulares, na cidade de Petrolina, Pernambuco. No presente trabalho, foram analisados a abordagem do histórico, os sinais clínicos, o diagnóstico laboratorial e o tratamento. Foi utilizada a técnica de inoculação em camundongo na identificação da toxina botulínica, que se mostrou sensível neste trabalho.


Botulism is a poisoning caused by ingestion of potent toxins of Clostridium botulinum, causing upward flaccid quadriplegia in dogs. There are seven types of botulism: A, B, C (?, ?), D, E, F and G, with the predominance of C and D in animals, attributed to the ingestion of raw or decaying meat. Although considered rare in most developed countries, it can still be found with some frequency. This article reports ten cases of canine type C botulism, originating from urban and rural areas and seen between the months of May 2009 and October 2010 in private veterinary practices in the city of Petrolina, Pernambuco, Brazil. The approach of anamnesis, clinical signs, and laboratory diagnosis were analyzed. The technique of inoculation in mice was used for the identification of the botulinum toxin, which has shown to be sensitive in this work.


El botulismo es una intoxicación que suele producirse por el consumo de carne en mal estado o cruda, provocada por la ingestión de potentes toxinas de Clostridium botulinum, que en los perros causan una tetraparesia fláxida ascendente. En los animales existen siete tipos de toxinas: A, B, C (?,?), D, E, F e G, con predominio de la C y D. A pesar de ser considerada una enfermedad de aparición rara en los países desarrollados, la misma puede ser encontrada con cierta frecuencia. Este trabajo relata diez casos de botulismo canino tipo C, originados en áreas urbanas y rurales, que fueron atendidos entre mayo de 2009 y octubre de 2010 en clínicas particulares de la ciudad de Petrolina, Pernambuco, Brasil. En este trabajo se analizaron el histórico del paciente, sus signos clínicos, exámenes de laboratorio y el tratamiento. Se utilizó una técnica de inoculación en ratones, que fue sensible para la identificación de la toxina botulínica.


Subject(s)
Animals , Dogs , Botulism , Diagnosis , Poisoning/complications , Dogs/classification
13.
Braz J Microbiol ; 44(1): 133-7, 2013.
Article in English | MEDLINE | ID: mdl-24159295

ABSTRACT

The objective of this study was to detect C. difficile A/B toxins and to isolate strains of C. perfringens and C. difficile from diarrheic and non-diarrheic dogs in Brazil. Stool samples were collected from 57 dogs, 35 of which were apparently healthy, and 22 of which were diarrheic. C. difficile A/B toxins were detected by ELISA, and C. perfringens and C. difficile were identified by multiplex PCR. C. difficile A/B toxins were detected in 21 samples (36.8%). Of these, 16 (76.2%) were from diarrheic dogs, and five (23.8%) were from non-diarrheic dogs. Twelve C. difficile strains (21.1%) were isolated, of which ten were A(+)B(+) and two were A(-)B(-). All non-toxigenic strains were isolated from non-diarrheic animals. The binary toxin gene cdtB was found in one strain, which was A(+)B(+) and was derived from a non-diarrheic dog. C. perfringens strains were isolated from 40 samples (70.2%). Of these, 18 (45%) were from the diarrheic group, and 22 (55%) belonged to the non-diarrheic group. All isolates were classified as C. perfringens type A and there was an association between the detection of the cpe gene and the presence of diarrhea. Interestingly, ten strains (25%) were positive for the presence of the cpb2 gene. The high rate of detection of the A/B toxins in non-diarrheic dogs suggests the occurrence of subclinical disease in dogs or carriage of its toxins without disease. More studies are needed to elucidate the epidemiology of C. difficile and C. perfringens in dogs and to better our understanding of C. difficile as a zoonotic agent. This is the first study to report the binary toxin gene in C. difficile strains isolated from dogs in Brazil.

14.
PLoS One ; 8(7): e69692, 2013.
Article in English | MEDLINE | ID: mdl-23936080

ABSTRACT

Bovine botulism is a fatal disease that is caused by botulinum neurotoxins (BoNTs) produced by Clostridium botulinum serotypes C and D and that causes great economic losses, with nearly 100% lethality during outbreaks. It has also been considered a potential source of human food-borne illness in many countries. Vaccination has been reported to be the most effective way to control bovine botulism. However, the commercially available toxoid-based vaccines are difficult and hazardous to produce. Neutralizing antibodies targeted against the C-terminal fragment of the BoNT heavy chain (HC) are known to confer efficient protection against lethal doses of BoNTs. In this study, a novel recombinant chimera, consisting of Escherichia coli heat-labile enterotoxin B subunit (LTB), a strong adjuvant of the humoral immune response, fused to the HC of BoNT serotypes C and D, was produced in E. coli. Mice vaccinated with the chimera containing LTB and an equivalent molar ratio of the chimera without LTB plus aluminum hydroxide (Al(OH)3) developed 2 IU/mL of antitoxins for both serotypes. Guinea pigs immunized with the recombinant chimera with LTB plus Al(OH)3 developed a protective immune response against both BoNT/C (5 IU/mL) and BoNT/D (10 IU/mL), as determined by a mouse neutralization bioassay with pooled sera. The results achieved with guinea pig sera fulfilled the requirements of commercial vaccines for prevention of botulism, as determined by the Brazilian Ministry of Agriculture, Livestock and Food, Supply. The presence of LTB was essential for the development of a strong humoral immune response, as it acted in synergism with Al(OH)3. Thus, the vaccine described in this study is a strong candidate for the control of botulism in cattle.


Subject(s)
Botulism/immunology , Clostridium botulinum type C/immunology , Clostridium botulinum type D/immunology , Vaccines, Synthetic/immunology , Aluminum Hydroxide/immunology , Amino Acid Sequence , Animals , Antibodies, Bacterial/immunology , Antitoxins/immunology , Bacterial Toxins/genetics , Bacterial Toxins/immunology , Bacterial Toxins/metabolism , Base Sequence , Blotting, Western , Botulinum Toxins/genetics , Botulinum Toxins/immunology , Botulinum Toxins/metabolism , Botulism/prevention & control , Botulism/veterinary , Cattle , Drug Evaluation , Enterotoxins/genetics , Enterotoxins/immunology , Enterotoxins/metabolism , Enzyme-Linked Immunosorbent Assay , Escherichia coli Proteins/genetics , Escherichia coli Proteins/immunology , Escherichia coli Proteins/metabolism , Female , Guinea Pigs , Mice , Molecular Sequence Data , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/immunology , Recombinant Fusion Proteins/metabolism , Vaccination/methods , Vaccines, Synthetic/administration & dosage , Vaccines, Synthetic/standards
15.
Braz. J. Microbiol. ; 44(1): 133-137, 2013. tab
Article in English | VETINDEX | ID: vti-7982

ABSTRACT

The objective of this study was to detect C. difficileA/B toxins and to isolate strains of C. perfringensand C. difficile from diarrheic and non-diarrheic dogs in Brazil. Stool samples were collected from 57 dogs, 35 of which were apparently healthy, and 22 of which were diarrheic. C. difficileA/B toxins were detected by ELISA, and C. perfringensand C. difficilewere identified by multiplex PCR. C. difficileA/B toxins were detected in 21 samples (36.8%). Of these, 16 (76.2%) were from diarrheic dogs, and five (23.8%) were from non-diarrheic dogs. Twelve C. difficile strains (21.1%) were isolated, of which ten were A+B+and two were A-B-. All non-toxigenic strains were isolated from non-diarrheic animals. The binary toxin gene cdtBwas found in one strain, which was A+B+and was derived from a non-diarrheic dog. C. perfringensstrains were isolated from 40 samples (70.2%). Of these, 18 (45%) were from the diarrheic group, and 22 (55%) belonged to the non-diarrheic group. All isolates were classified as C. perfringenstype A and there was an association between the detection of the cpegene and the presence of diarrhea. Interestingly, ten strains (25%) were positive for the presence of the cpb2gene. The high rate of detection of the A/B toxins in non-diarrheic dogs suggests the occurrence of subclinical disease in dogs or carriage of its toxins without disease. More studies are needed to elucidate the epidemiology of C. difficileand C. perfringensin dogs and to better our understanding of C. difficileas a zoonotic agent. This is the first study to report the binary toxin gene in C. difficilestrains isolated from dogs in Brazil.(AU)


Subject(s)
Animals , Toxicology , Diarrhea/pathology , Colitis/pathology , Enteritis/pathology , Dogs/classification
16.
Semina ciênc. agrar ; 34(1): 355-358, 2013.
Article in Portuguese | LILACS-Express | VETINDEX | ID: biblio-1499113

ABSTRACT

This work aims to describe an outbreak of botulism in poultry (Gallus gallus domesticus), in Pancas City, Espírito Santo State, Brazil. The birds were derived from a subsistence property with approximately 150 domestic fowls. The owner reported finding some birds lying on the floor and others exhibiting limited mobility. Ten animals in agony were euthanized, necropsied and had their blood serum collected. About 72 hours after the onset of clinical signs, 144 (96%) animals of the creation had already died. No postmortem changes were found at necropsies. Clostridium botulinum type C toxins were detected in the sera of euthanized chickens by mice neutralization tests. The observation of clinical signs, absence of post-mortem lesions and the result of mouse neutralization tests confirmed the diagnosis of botulism caused by C. botulinum type C. Unfortunately, the source of toxins was not found.


O presente trabalho tem como objetivo descrever um surto de botulismo em aves domésticas (Gallus gallus domesticus), ocorrido na cidade de Pancas, estado do Espírito Santo, Brasil. As aves eram oriundas de uma propriedade de subsistência com aproximadamente 150 frangos domésticos. O proprietário relatou encontrar algumas aves caídas no chão e outras apresentando dificuldade de locomoção. Dez animais em estado agônico foram eutanasiados, submetidos à necropsia e coleta de soro sanguíneo. Em 72 horas após o início dos sinais clínicos, 144 (96%) animais da criação já haviam vindo a óbito. Nenhuma alteração post-mortem foi encontrada durante a necropsia. Foi possível detectar a toxina botulínica tipo C por soroneutralização em camundongos no soro dos animais eutanasiados. A observação da sintomatologia clínica, ausência de lesões à necropsia e o resultado da soroneutralização permitem confirmar o diagnóstico de botulismo por Clostridium botulinum tipo C.

17.
Semina Ci. agr. ; 34(1): 355-358, 2013.
Article in Portuguese | VETINDEX | ID: vti-763027

ABSTRACT

This work aims to describe an outbreak of botulism in poultry (Gallus gallus domesticus), in Pancas City, Espírito Santo State, Brazil. The birds were derived from a subsistence property with approximately 150 domestic fowls. The owner reported finding some birds lying on the floor and others exhibiting limited mobility. Ten animals in agony were euthanized, necropsied and had their blood serum collected. About 72 hours after the onset of clinical signs, 144 (96%) animals of the creation had already died. No postmortem changes were found at necropsies. Clostridium botulinum type C toxins were detected in the sera of euthanized chickens by mice neutralization tests. The observation of clinical signs, absence of post-mortem lesions and the result of mouse neutralization tests confirmed the diagnosis of botulism caused by C. botulinum type C. Unfortunately, the source of toxins was not found.


O presente trabalho tem como objetivo descrever um surto de botulismo em aves domésticas (Gallus gallus domesticus), ocorrido na cidade de Pancas, estado do Espírito Santo, Brasil. As aves eram oriundas de uma propriedade de subsistência com aproximadamente 150 frangos domésticos. O proprietário relatou encontrar algumas aves caídas no chão e outras apresentando dificuldade de locomoção. Dez animais em estado agônico foram eutanasiados, submetidos à necropsia e coleta de soro sanguíneo. Em 72 horas após o início dos sinais clínicos, 144 (96%) animais da criação já haviam vindo a óbito. Nenhuma alteração post-mortem foi encontrada durante a necropsia. Foi possível detectar a toxina botulínica tipo C por soroneutralização em camundongos no soro dos animais eutanasiados. A observação da sintomatologia clínica, ausência de lesões à necropsia e o resultado da soroneutralização permitem confirmar o diagnóstico de botulismo por Clostridium botulinum tipo C.

18.
Semina Ci. agr. ; 34(1): 355-358, 2013.
Article in Portuguese | VETINDEX | ID: vti-471345

ABSTRACT

This work aims to describe an outbreak of botulism in poultry (Gallus gallus domesticus), in Pancas City, Espírito Santo State, Brazil. The birds were derived from a subsistence property with approximately 150 domestic fowls. The owner reported finding some birds lying on the floor and others exhibiting limited mobility. Ten animals in agony were euthanized, necropsied and had their blood serum collected. About 72 hours after the onset of clinical signs, 144 (96%) animals of the creation had already died. No postmortem changes were found at necropsies. Clostridium botulinum type C toxins were detected in the sera of euthanized chickens by mice neutralization tests. The observation of clinical signs, absence of post-mortem lesions and the result of mouse neutralization tests confirmed the diagnosis of botulism caused by C. botulinum type C. Unfortunately, the source of toxins was not found.


O presente trabalho tem como objetivo descrever um surto de botulismo em aves domésticas (Gallus gallus domesticus), ocorrido na cidade de Pancas, estado do Espírito Santo, Brasil. As aves eram oriundas de uma propriedade de subsistência com aproximadamente 150 frangos domésticos. O proprietário relatou encontrar algumas aves caídas no chão e outras apresentando dificuldade de locomoção. Dez animais em estado agônico foram eutanasiados, submetidos à necropsia e coleta de soro sanguíneo. Em 72 horas após o início dos sinais clínicos, 144 (96%) animais da criação já haviam vindo a óbito. Nenhuma alteração post-mortem foi encontrada durante a necropsia. Foi possível detectar a toxina botulínica tipo C por soroneutralização em camundongos no soro dos animais eutanasiados. A observação da sintomatologia clínica, ausência de lesões à necropsia e o resultado da soroneutralização permitem confirmar o diagnóstico de botulismo por Clostridium botulinum tipo C.

19.
Semina ciênc. agrar ; 34(1): 355-358, 2013.
Article in Portuguese | LILACS-Express | VETINDEX | ID: biblio-1433401

ABSTRACT

This work aims to describe an outbreak of botulism in poultry (Gallus gallus domesticus), in Pancas City, Espírito Santo State, Brazil. The birds were derived from a subsistence property with approximately 150 domestic fowls. The owner reported finding some birds lying on the floor and others exhibiting limited mobility. Ten animals in agony were euthanized, necropsied and had their blood serum collected. About 72 hours after the onset of clinical signs, 144 (96%) animals of the creation had already died. No postmortem changes were found at necropsies. Clostridium botulinum type C toxins were detected in the sera of euthanized chickens by mice neutralization tests. The observation of clinical signs, absence of post-mortem lesions and the result of mouse neutralization tests confirmed the diagnosis of botulism caused by C. botulinum type C. Unfortunately, the source of toxins was not found.


O presente trabalho tem como objetivo descrever um surto de botulismo em aves domésticas (Gallus gallus domesticus), ocorrido na cidade de Pancas, estado do Espírito Santo, Brasil. As aves eram oriundas de uma propriedade de subsistência com aproximadamente 150 frangos domésticos. O proprietário relatou encontrar algumas aves caídas no chão e outras apresentando dificuldade de locomoção. Dez animais em estado agônico foram eutanasiados, submetidos à necropsia e coleta de soro sanguíneo. Em 72 horas após o início dos sinais clínicos, 144 (96%) animais da criação já haviam vindo a óbito. Nenhuma alteração post-mortem foi encontrada durante a necropsia. Foi possível detectar a toxina botulínica tipo C por soroneutralização em camundongos no soro dos animais eutanasiados. A observação da sintomatologia clínica, ausência de lesões à necropsia e o resultado da soroneutralização permitem confirmar o diagnóstico de botulismo por Clostridium botulinum tipo C.

20.
Braz. j. microbiol ; Braz. j. microbiol;44(1): 133-137, 2013. tab
Article in English | LILACS | ID: lil-676895

ABSTRACT

The objective of this study was to detect C. difficileA/B toxins and to isolate strains of C. perfringensand C. difficile from diarrheic and non-diarrheic dogs in Brazil. Stool samples were collected from 57 dogs, 35 of which were apparently healthy, and 22 of which were diarrheic. C. difficileA/B toxins were detected by ELISA, and C. perfringensand C. difficilewere identified by multiplex PCR. C. difficileA/B toxins were detected in 21 samples (36.8%). Of these, 16 (76.2%) were from diarrheic dogs, and five (23.8%) were from non-diarrheic dogs. Twelve C. difficile strains (21.1%) were isolated, of which ten were A+B+and two were A-B-. All non-toxigenic strains were isolated from non-diarrheic animals. The binary toxin gene cdtBwas found in one strain, which was A+B+and was derived from a non-diarrheic dog. C. perfringensstrains were isolated from 40 samples (70.2%). Of these, 18 (45%) were from the diarrheic group, and 22 (55%) belonged to the non-diarrheic group. All isolates were classified as C. perfringenstype A and there was an association between the detection of the cpegene and the presence of diarrhea. Interestingly, ten strains (25%) were positive for the presence of the cpb2gene. The high rate of detection of the A/B toxins in non-diarrheic dogs suggests the occurrence of subclinical disease in dogs or carriage of its toxins without disease. More studies are needed to elucidate the epidemiology of C. difficileand C. perfringensin dogs and to better our understanding of C. difficileas a zoonotic agent. This is the first study to report the binary toxin gene in C. difficilestrains isolated from dogs in Brazil.


Subject(s)
Animals , Dogs , Clinical Laboratory Techniques , Clostridium perfringens , Clostridioides difficile/genetics , Clostridioides difficile/isolation & purification , Diarrhea/genetics , Enterocolitis, Pseudomembranous , Fecal Impaction/genetics , Polymerase Chain Reaction , Bacterial Toxins/genetics , Bacterial Toxins/isolation & purification , Diagnosis , Immunoassay , Methods , Spores, Bacterial , Virulence
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