Iveraldo S. Dutra

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 botulinum proteolytic type B spores and their toxins. Results demonstrated that the harsh processing conditions, high salt concentration, relative high temperature, a(w) 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.

Surtos de botulismo em bovinos no Brasil associados à ingestão de água contaminada

Botulism in cattle occurs by ingestion of botulinum toxin C and/or D. Seven outbreaks of the poisoning in the states of Mato Grosso do Sul and Sao Paulo are reported. The clinical and pathological data, epidemiology and laboratory findings indicate a possible intake of the toxin through contaminated water. The average mortality rate was 20.1%, with 99.2% lethality and 31.62% morbidity. From about 9,000 cattle envolved in the outbreaks, 2,844 animals died, predominantly with a hyperacute and acute clinical picture. The high morbidity rates were observed within a short period and affected all categories of cattle, with a clinical and pathological picture characterized by paresis and paralysis of the muscles of locomotion, swallowing and mastigation, with abscence of gross lesions at post-mortem examination. The outbreaks were related to the presence of decomposed animal carcasses or vegetal material in the drinking water. Botulinum toxins C and/or D were detected in water samples, viscera and blood serum of a considerable number of materials examined.

Botulismo em bovinos de corte e leite alimentados com cama de frango

Outbreaks of botulism caused by type C and D of the botulinum toxin are frequent in Brazil, and are associated with bone chewing and ingestion of contaminated food and water. This paper reports the epidemiological, clinical, pathological and laboratorial aspects of 7 outbreaks of botulism in beef and dairy cattle fed with poultry litter, which occurred in the states of Sao Paulo and Minas Gerais, 1989-2000. Five outbreaks occurred in beef cattle herds, raised in confinement or under pasture conditions and supplemented with poultry litter, and 2 outbreaks occurred in dairy farms. From o total of 1,535 cattle supplemented regularily with poultry litter 455 animals (29.64%) died within 2 to 4 weeks. Morbidity and mortality varied from 3.47 to 100% in the 7 outbreaks. In one of the farms the lethality was 60.52%, and in others more than 88.43%, reaching 100% in three farms. Clinical signs were progressive paralysis, difficulties in moving, decubitus, normal alertness, decreased muscular tonus of tongue and tail, sialorrhoe and dyspnoe. At post-mortem examination of 30 cattle no noteable changes were observed. Spores of Clostridium botulinum were found in poultry litter samples collected on 7 farms. In liver, ruminal and intestinal fluid samples from 30 necropsied cattle botulinum toxin of type C (5) and D (9) or of the CD complex (1) were found in at least one of the samples collected from 15 animals, which confirms the clincial, pathological and epidemiological diagnosis of botulism.

Pré-compostagem de cadáveres de bovinos acometidos pelo botulismo

Five carcasses of cattle that died from botulism were composted in individual isolated beds with the purpose of monitoring the effect on spores of Clostridium botulinum and of botulinum toxin. The diagnosis of botulinum poisoning was based on clinical and pathological, epidemic and/or laboratorial characteristics. Of the five bovines with clinical signs of chronic botulism, three were affected by the type D toxin, one by the CD complex, and one was negative for toxin and spores of C. botulinum in the viscera using a mouse bioassay. The composting process was carried out in beds with carcasses cut up and mixed with sawdust as a carbon source. They were kept anaerobic under a plastic sheet for 50 days. The temperature of the beds was monitored during the period and oscillated from 40.5 to 52.4oC. After this time, soft tissue had complete decomposed with a significant loss of weight (26.5-44.5%). The remaining bones no longer contained spores or botulinum toxin in the 5 samples taken from each carcass. Only 2 of the 200 samples of homogenized remaining (40 for each carcass) contained spores of C. botulinum type C. All samples were negative for botulinum toxin using the mouse bioassay. Composting carcasses of cattle that died from botulism eliminated botulinum spores and toxin from the remains.

Febre catarral maligna em bovinos do Mato Grosso do Sul e de São Paulo

The epidemiology, clinical signs, necropsy findings and histopathology of 6 cases of malignant catarrhal fever (MCF) in cattle from five farms located in the states of Mato Grosso do Sul and Sao Paulo, Brazil are described. The disease occurred in cattle of both sexes, ranging from 4-months to 11 years of age. Morbidity rates were 0.25%-6.6% and lethality was 100%. Clinical courses were acute in six cases (2-3 days) and chronic in one (3 months). Presumptive diagnosis of MCF in the cases in this report were made by clinical signs and necropsy findings and comfirmed by histopathological examination. Clinical signs in acute cases were fever, mucopurulent nasal discharge, corneal opacity, drooling and ulcers in several mucosal surfaces and neurologic disturbances. The chronic case had corneal opacity and neurologic signs. Necropsy findings included hyperemia and diphteric lesions in several epithelial surfaces and histopathology consisted of vasculitis, multifocal disseminated foci of mononuclear cell infiltrate, and necrosis of epithelial surfaces.

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

The occurrence and distribution of Clostridium botulinum spores and toxins type C and D in 300 ponds, used by cattle for drinking on 130 farms located in 12 municipalities of the Rio Araguaia valley, State of Goias, Brazil, was evaluated. The presence of spores was determined indirectly by cultivation in culture medium, followed by inoculation and neutralization in mice of samples of the sediment from the bottom of the raining ponds, from superficial soil and from cattle feces collected to its circuit. The toxin presence was evaluated directly by inoculation in mice of the filtered sediment of the ponds, followed by the neutralization in mice with antitoxins C and D. The presence of C. botulinum spores was significantly more frequent (p<0,05) in the cattle feces (31%), when compared with the results of the superficial soil samples (19%) and the sediments (10%). Botulinum toxins of type C and D or classified as belonging to the CD compound were detected in 6 samples (2%) of the 300 ponds. Of the 130 worked farms, in 122 (93,85%) ponds Clostridium botulinum spores or toxins were found in at least one of the researched variables, whilst ponds on only 8 (6,15%) farms did not present any contamination. Age and depth of the ponds were associated with the frequency of detection of botulinum spores and toxins. The older and shallower the ponds were, the larger was the frequency of isolation of the spores and toxins. The contamination of the ponds in the Araguaia valley with Clostridium botulinum spores and toxins type C and D demonstrates the permanent and growing potential risk for the occurrence of botulism in cattle through drinking water.

Treponema denticola na microbiota da periodontite bovina

Periodontitis in cattle is an infectious purulent progressive disease associated with strict anaerobic subgingival biofilm and is epidemiologically related to soil management at several locations of Brazil. This study aimed to detect Treponema species in periodontal pockets of cattle with lesions deeper than 5mm in the gingival sulcus of 6 to 24-month-old animals considered periodontally healthy. We used paper cones to collect the materials, after removal of supragingival plaques, and kept frozen (at -80°C) up to DNA extraction and polymerase chain reaction (PCR) using T. amylovorum, T. denticola, T. maltophilum, T. medium and T. vincentii primers. In periodontal pocket, it was possible to identify by PCR directly, the presence of Treponema amylovorum in 73% of animals (19/26), T. denticola in 42.3% (11/26) and T. maltophilum in 54% (14/26). Among the 25 healthy sites, it was possible to identify T. amylovorum in 18 (72%), T. denticola in two (8%) and T. maltophilum in eight (32%). Treponema medium and T. vincentii were not detected over all 51 evaluated samples. The presence of Treponema amylovorum, T. maltophilum and, in particular, the widely recognized T. denticola in subgingival microflora brings an original and potencially important contribution in studies of the bovine periodontitis.

Vaccination protocol and bacterial strain affect the serological response of beef calves against blackleg

The serological response of beef calves was evaluated with different vaccination regimens against blackleg, using an official strain (MT) and a field-collected strain of Clostridium chauvoei as antigens. Sixty calves were randomly allocated to four different groups and were submitted to distinct vaccination protocols with a commercial polyvalent vaccine. Group G1 was first vaccinated at four months of age and a booster shot was given after weaning, at eight months. Group G2 was given the first dose at eight months and a booster shot 30 days later. Group G3 was vaccinated only once at eight months and the control group was not vaccinated. These alternative vaccination regimens were proposed in an effort to adequately protect cattle under open-field farming conditions. Serological evaluations were made by Elisa at 4, 8, 9 and 10 months of age. Both groups receiving booster shots had a significantly increased serological response 30 days later. However, the serum IgG levels against C. chauvoei were significantly higher in the calves that were first vaccinated at four months. At 10 months, the two booster shot groups (G1 and G2) had similar serological responses, while the calves that were treated with a single dose of vaccine at weaning (G3) had a response that was similar to that of the control group. The serological response of the calves was significantly inferior at several of the evaluation times when the field strain of the bacteria was used as a challenge antigen instead of the official MT strain. The serological response of calves that are vaccinated twice was found to be satisfactory, independent of the first injection being made at four or eight months of age. It was also concluded that it would be useful to include local bacterial strains in commercial vaccine production.

Immunoprophylactic strategies against enterotoxemia caused by Clostridium perfringens type D in goats

The serological response to an experimental vaccine against Clostridium perfringens type D enterotoxemia was evaluated in four groups of goats. Group 1 received colostrum from unvaccinated cows and no vaccine. Groups 2, 3 and 4 received colostrum from vaccinated cows. In addition, Groups 3 and 4 received a vaccine dose at 80 days of age, and Group 4 received a second vaccine dose at 120 days of age. Serum antibody levels were determined by ELISA in cows before and after calving, and in goats at 3, 80, 120 and 160 days of age. No significant difference in serum antibody levels was observed between vaccinated and unvaccinated cows, or between the four groups of goats evaluated at 3 days of life. Groups 3 and 4 presented mean antibody titers of 0.6 and 1.1 IU/ml, respectively, 40 days after first vaccination. The vaccine response of Group 4 was 1.8 IU/ml 40 days after the booster dose and was higher than that observed for Group 3 (0.2 IU/ml). Thus, in the proposed regimen the use of heterologous colostrum did not induce passive immunization in goat kids. However, first vaccination and a booster dose after 40 days triggered satisfactory antibody levels.

Naturally acquired antibodies against Clostridium perfringens epsilon toxin in goats

Clostridium perfringens type D-producing epsilon toxin is a common cause of death in sheep and goats worldwide. Although anti-epsilon toxin serum antibodies have been detected in healthy non-vaccinated sheep, the information regarding naturally acquired antibodies in ruminants is scanty. The objective of the present report was to characterize the development of naturally acquired antibodies against C. perfringens epsilon toxin in goats. The levels of anti-epsilon toxin antibodies in blood serum of goat kids from two different herds were examined continuously for 14 months. Goats were not vaccinated against any clostridial disease and received heterologous colostrums from cows that were not vaccinated against any clostridial disease. During the survey one of these flocks suffered an unexpectedly severe C. perfringens type D enterotoxemia outbreak. The results showed that natural acquired antibodies against C. perfringens epsilon toxin can appear as early as 6 weeks in young goats and increase with the age without evidence of clinical disease. The enterotoxemia outbreak was coincident with a significant increase in the level of anti-epsilon toxin antibodies.