Myonecrosis by Clostridium septicum in a dog, diagnosed by a new multiplex-PCR

Clostridial myositis is an acute, generally fatal toxemia that is considered to be rare in pet animals. The present report describes an unusual canine clostridial myositis that was diagnosed by a new multiplex-PCR (mPCR) designed for simultaneous identification of Clostridium sordellii, Clostridium septicum, Clostridium perfringens type A, Clostridium chauvoei, and Clostridium novyi type A. A ten-month-old male Rottweiler dog, that had displayed lameness and swelling of the left limb for 12 h, was admitted to a veterinary hospital. The animal was weak, dyspneic and hyperthermic, and a clinical examination indicated the presence of gas and edema in the limb. Despite emergency treatment, the animal died in only a few minutes. Samples of muscular tissue from the necrotic area were aseptically collected and plated onto defibrinated sheep blood agar (5%) in anaerobic conditions. Colonies suggestive of Clostridium spp. were submitted to testing by multiplex-PCR. Impression smears of the tissues, visualized with Gram and also with panoptic stains, revealed long rod-shaped organisms, and specimens also tested positive using the fluorescent antibody technique (FAT). The FAT and mPCR tests enabled a diagnosis of C. septicum myonecrosis in the dog. (C) 2012 Elsevier Ltd. All rights reserved.

An unusual gangrenous goat mastitis caused by Staphylococcus aureus, Clostridium perfringens and Escherichia coli co-infection

Relata-se, pela primeira vez no Brasil, a ocorrência de mastite gangrenosa caprina atípica causada pela co-infecção por Staphylococcus aureus, Clostridium perfringens e Escherichia coli em uma cabra da raça Boer, na segunda semana de lactação. Descrevem-se os achados clínicos, os procedimentos de diagnóstico microbiológico e a conduta terapêutica.

Naturally acquired antibodies against Clostridium perfringens epsilon toxin in goats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Inhibitors of Clostridium histolyticum colagenase

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ocorrência e distribuição de esporos de Clostridium botulinum tipos C e D em áreas de criação de búfalos na Baixada Maranhense

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Esporos e toxinas de Clostridium botulinum dos tipos C e D em cacimbas no Vale do Araguaia, Goiás

Foram avaliadas a ocorrência e distribuição de esporos e toxinas de Clostridium botulinum tipos C e D em 300 cacimbas empregadas como bebedouro de bovinos em 130 propriedades rurais localizadas em 12 municípios do Vale do Araguaia, Estado de Goiás. A presença de esporos foi determinada indiretamente pelo cultivo em meio de cultura, seguido da inoculação e neutralização em camundongo das amostras de sedimento do interior das cacimbas, e do solo superficial e fezes de bovinos, coletadas ao seu redor. A presença de toxina foi avaliada diretamente pela inoculação em camundongo do sedimento filtrado das cacimbas, também seguida da neutralização em camundongo com antitoxinas C e D. A presença de esporos de C. botulinum foi significativamente maior (p<0,05) nas fezes de bovinos (31%), quando comparadas com os resultados das amostras de solo superficial (19%) e dos sedimentos (10%). Foram detectadas toxinas botulínicas dos tipos C, D, ou classificadas como pertencentes ao complexo CD, em seis amostras (2%) das 300 cacimbas. Das 130 propriedades trabalhadas, em 122(93,85%) foram encontrados esporos ou toxinas de Clostridium botulinum em pelo menos uma das variáveis pesquisadas, enquanto somente 8(6,15%) não apresentaram qualquer contaminação A idade e profundidade das cacimbas estiveram associadas com a freqüência de detecção de esporos e toxinas. Assim, quanto mais velhas e rasas, maior a freqüência do isolamento de esporos e toxinas. A contaminação das cacimbas do Vale do Araguaia goiano com esporos e toxinas do Clostridium botulinum tipos C e D demonstra o risco potencial permanente e crescente para a ocorrência da intoxicação botulínica de origem hídrica nos bovinos.

Microbiological analyses of poultry litter used for ruminant feeding

The aims of this study were to estimate the changes in total bacterial counts (TBC) in poultry litter samples, consisting of rice hulls, after storage, and to identify pathogenic bacteria. For the countings Plate Count agar (Difco) was used. Enrichment and selective media such as blood agar, MacConkey, Baird Parker, brain and heart agar, and egg yolk solid media, and cooked meat and brain and heart infusion, incubated under aerobic or anaerobic conditions were used to isolate Staphylococcus aureus, Salmonella sp, Clostridium perfringens, C. botulinum, C. chauvoei, Campylobacter sp, Escherichia coli, and Corynebacterium sp. Litter samples were collected from the houses of the Veterinary School experimental aviary. A fully randomized experimental design was used with four treatments and four replications, for a total of 16 samples. A decrease in TBC was detected when treatment T1 (zero days of storage) was compared with treatments T2 (14 days of storage). on the other hand the treatments T3 (28 days of storage) and T4 (42 days of storage) presented significantly superior counting in relation to treatment T1. Some pathogenic bacteria of Enterobacteriaceae such as Escherichia coil, Proteus, Arizona, Providencia, Edwardsiella, as well as Staphylococcus aureus, S. epidermidis, different species of genus Clostridium as C. perfringens, C. sordelli, C. chauvoei, C. tetani and C. novyi as well as some strains of Corynebacterium pyogenes were isolated.

GENETIC INTERRELATIONSHIPS OF PROTEOLYTIC CLOSTRIDIUM-BOTULINUM TYPE-A, TYPE-B, AND TYPE-F AND OTHER MEMBERS OF THE CLOSTRIDIUM-BOTULINUM COMPLEX AS REVEALED BY SMALL-SUBUNIT RIBOSOMAL-RNA GENE-SEQUENCES

The phylogenetic interrelationships of members of the Clostridium botulinum complex of species was investigated by direct sequencing of their 16S rRNA genes. Comparative analysis of the 16S rRNA sequences demonstrated the presence of four phylogenetically distinct lineages corresponding to: i) proteolytic C. botulinum types Al B, and F, and C. sporogenes, ii) saccharolytic types B, E and F, iii) types C and D and C. novyi type A, and iv) type G and C. subterminale. The phylogenetic groupings obtained from the 16S rRNA were in complete agreement with the four divisions recognised within the 'species complex' on the basis of phenotypic criteria.

Evaluation of survival of Staphylococcus aureus and Clostridium botulinum in charqui meats

Charqui meats were prepared in laboratory conditions in order to carry out experiments to observe the possibility of development of enterotoxigenic Staphylococcus aureus and Clostridium bottilinum proteolytic type B spores and their toxins. Results demonstrated that the harsh processing conditions, high salt concentration, relative high temperature, a, values, inhibited the growth of both bacteria. Under our experimental conditions, S. aureus would survive throughout the sequence of salting steps i.e. brine followed by rock salting and the sunshine drying step. However, at final a(w) value of 0.70-0.75 would create conditions to inhibit its development. The other experiment revealed that C. botulinum spores germination also was impaired because of these low a(w) values. Under these conditions, charqui meats revealed to be safe products in relation to toxins from both enterotoxigenic S. aureus and C. botulinum. (C) 2003 Elsevier B.V. Ltd. All rights reserved.

Neonatal immune response of Brazilian beef cattle to vaccination with Clostridium botulinum toxoids types C and D by indirect ELISA

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Botulism by Clostridium botulinum type C in goats associated with osteophagia

An outbreak of botulism was detected in goats in the semiarid region of Brazil. In a flock of 460 goats, 38 does were affected and 37 died. Kids and younger goats were not affected. The main clinical signs were flaccid tetraparesis leading to tetraplegia that was often accompanied by twisted neck, tongue paralysis, and muscle tremors. At the time of the visit, 4 out of 11 affected goats were recumbent. Ambulatory goats had uncoordinated and swaying gaits with hypometria and weakness, mainly of the hind limbs. Two recumbent and four ambulatory goats showed twisted neck. Two recumbent goats were euthanized and necropsied. Non-significant gross and histologic lesions were observed. Samples of the liver, gut and rumen content were collected from the two goats and examined for botulinum toxins using the mouse serum neutralization test. The three samples from one goat were positive for type C toxin. Marked osteophagia was observed when the goats had access to bones in the pasture, and the farmer mentioned that osteophagia was common among goats of the flock. A sample of the plant Hybantus ipecaconha, the most abundant forage available for the goats, contained 2800 mg/kg of Ca and 450 mg/kg of P. One soil sample contained 58.12 mg/kg of Ca and 2.02 mg/kg of P. It was concluded that in this outbreak, botulism was associated with osteophagia probably due to phosphorus deficiency. (C) 2012 Elsevier B.V. All rights reserved.

Study of the presence of the spores of Clostridium botulinum in honey in Brazil

The isolation of Clostridium botulinum from honey samples is described. Botulism is characterized as an intoxication provoked by ingestion of contaminated foods with this toxin. Infant botulism happens by the ingestion of spores of C. botulinum together with food that in special conditions of the intestinal tract, such as those present in babies of less than 1 year old, will allow the germination and colonization of the intestine with production and absorption of botulinic toxin. The samples were subjected to dilution and to a thermal shock and cultivated in modified CMM (Difco). Cultures were subjected to Gram smears and toxicity tests in mice. The toxic cultures were purified in RFCA (Oxoid) plates and incubated in anaerobic jars. Positive samples were typed using the mouse assay neutralization test. From the 85 honey samples analyzed, six were positive for C. botulinum (7.06%), and identified as producers of type A, B, and D toxins.

Intoxicação natural por Clostridium botulinum tipo C: em grupo de aves domésticas

Clostridium botulinum causative of toxic infections due to toxin ingestion previously formed, occur in several ingestion species, mainly birds. In a poultry farm, located at São Paulo State, more than 3,000 birds have been attacked by this toxic infections, when the birds showed motionless, loss of weight, accelerated, breathing, resulting in death of the birds. The blood harvest for attainment of the serum and later necropsy was carried out throwgh the following samples: liver, gizzard, crop, feed, water and litter. After that, the toxin was extracted by gelatin-phosphate buffer and inoculation in mice, isolation of the agent in Blood. Ágar and Reinforced Clostridium Ágar and neutralization of toxin determine its type. The inoculation in mice showed positive results in samples of liver, gizzad, crop and symptoms like was waist, and death of the birds by limp paralysis. The colonies that have been isolated, suspected of Clostridium botulinum showed expected results and further analysis revealed positive results to botulinical toxin type C. The farms have to pay attention in the routine jobs, choicer, and elimination of carcass because they are essential to keep this problem away.