Effect of double-premilking teat disinfection protocols on bacterial counts on teat skin of cows and milker gloves in a free-stall-housed dairy herd.

This research communication addresses the hypothesis that double premilking teat disinfection (DD) is more effective in reducing soiling and bacterial count on the cows' teat skin and milkers' gloves than conventional disinfection (CONV). The design was a 3 × 3 Latin square (three groups of cows and three treatments) with conventional teat disinfection (CONV, lactic acid application after forestripping), double teat disinfection using the same disinfectant (DD1D, lactic acid application before and after forestripping), and double teat disinfection using two different disinfectants (DD2D, application of lactic acid before and chlorine-based disinfectant after forestripping). All groups were assigned for six days for each treatment, and the evaluations and samples were collected on the last day, before and after treatment. We evaluated the teat cleanliness score (TCS), count of Gram-negative bacteria (coliforms and noncoliforms), Staphylococcus spp., Streptococcus spp., and total bacterial count (TBC) on the cows' teats and TBC on the milkers' gloves. TCS after premilking was lower in DD than CONV treatment. The reduction of Staphylococcus spp. count was greater in the DD treatment and tended to be higher in the DD2D. The TBC reduction on the cows' teats was greater in the DD treatments. The TBC on the milkers' gloves was lower for DD before and after premilking. In conclusion, DD can be an alternative for reducing some bacterial populations on cow teats and preventing the transmission of microorganisms between cows via the milkers' hands.

Spread analysis of glanders in the state of Piauí, northeastern Brazil.

Brazil is strategic in controlling neglected zoonoses, such as glanders, in its territory. Among the Brazilian states, Piauí is a strategic state for the spread of the disease in the country. The present study aimed to evaluate the spatial and temporal distribution of official cases of glanders in Piauí between 2015 and 2022. The glanders cases were located in the municipalities of the north and central-north mesoregions, mainly in Campo Maior, Teresina and Altos. The highest incidence risk (IR) occurred in of Altos (IR = 257.9), Sussuapara (IR = 158.4), and Teresina (IR = 157.7). A primary cluster was formed with a relative risk of 14.88 between 2019 and 2022, encompassing 34 municipalities in the north and central-north regions. In Piauí, glanders is well localized, with the potential for spread across borders. This is the first study demonstrating the distribution of reported cases of glanders in the state of Piauí.

Causes of fetal death in the Flemish cattle herd in Brazil.

Background and Aim: Flemish cattle in Brazil are on the brink of extinction and are found only in one herd in Lages, Santa Catarina State. This study aimed to uncover the reasons for the recurring abortions in the Flemish cattle herd. Materials and Methods: Seventeen Flemish fetuses underwent postmortem examinations, with samples collected for histopathology and microbiology culture tests, polymerase chain reaction (PCR) test for Neospora caninum, and reverse transcription-PCR (RT-PCR) test for bovine viral diarrhea virus (BVDV) from 2015 to 2020. Results: Of the 17 fetuses, N. caninum was the most common diagnosis and was found in 88% (15/17). One fetus (5.8%) had a coinfection with N. caninum and Citrobacter amalonaticus, leading to fibrinonecrotic pericarditis. All fetuses tested negative for BVDV by RT-PCR. Of the 107 dams tested by indirect immunofluorescence assay, 26 (25.2%) were anti-N. caninum seropositive, with 17 (65.4%) aborting and 5 (19.2%) having estrus repetition. Reverse transcription-PCR results showed that 9 (8.4%) of the serum samples collected from dams tested positive, which tested follow-up test 3 months later, indicating a BVDV transient infection. The factors that contributed to neosporosis included dogs' access to pastures and improper disposal of fetal remains, which made it easier for dogs to consume them. Conclusion: This study warns the occurrence of N. caninum as a cause of reproductive disorders that can lead to abortion in the studied Flemish cattle herd.

Detection of virulence genes and antimicrobial susceptibility profile of Listeria monocytogenes isolates recovered from artisanal cheese produced in the Southern region of Brazil.

Listeria monocytogenes is an opportunistic pathogen that causes listeriosis, a foodborne disease with low incidence but with high mortality rate in humans. This microorganism has been recovered from several dairy products, especially those produced with raw milk. The objective of this work was to investigate the presence of virulence genes, and also to define the antimicrobial susceptibility profile of L. monocytogenes isolates recovered from serrano artisanal cheese produced in Southern region of Brazil. Nine strains of L. monocytogenes (serotypes 1/2b and 4b) were evaluated through PCR to detect the presence of the virulence genes hly, inlA, inlC, inlJ, actA, plcB and iap, while antimicrobial susceptibility profile was determined via disk diffusion method. All strains exhibited the presence of the genes hly and plcB, whereas the other genes (iap, actA, inlA, inlC and inlJ) were only detected in eight strains. We verified that all strains were resistant to at least one antimicrobial agent and three of them showed multidrug resistance. These findings demonstrated the serrano artisanal cheese offers risks to consumers' health and point to a need of adaptations and monitoring of manufacturing process of this food, in order to prevent the dissemination of L. monocytogenes.

Salivary ß-glucosidase as a direct factor influencing the occurrence of halitosis.

ß-Glucosidases are enzymes present in all living organisms, playing a pivotal role in diverse biological processes. These enzymes cleave ß-glycosidic bonds between carbohydrates, or between a carbohydrate and a non-carbohydrate moiety, which may result in the liberation of volatile aglycones. Released compounds execute diverse physiological roles, while the industry takes advantage of exogenously added ß-glucosidases for aroma enrichment during food and beverage production. ß-Glucosidase enzymatic activity has been reported in human saliva and given the fact that these enzymes are involved in aroma release, we investigated here the correlation between ß-glucosidase activity in human saliva and the occurrence of halitosis. Measurement of salivary enzyme activity of 48 volunteers was performed using p-nitrophenyl-ß-d-glucopyranoside as substrate. Each volunteer was clinically evaluated by a dental surgeon and clinical and laboratorial data were statistically analyzed. Gas-chromatography of saliva headspace allowed the analysis of the direct role of exogenous ß-glucosidase on aromatic /volatile profile of saliva samples. The data demonstrated a positive correlation between halitosis and enzymatic activity, suggesting that the enzyme exerts a direct role in the occurrence of bad breath. Gas-chromatography analysis demonstrated that exogenously added enzyme led to the alteration of volatile organic content, confirming a direct contribution of ß-glucosidase activity on saliva volatile compounds release. Although halitosis is a multifactorial condition, the complete understanding of all governing factors may allow the development of more effective treatment strategies. Such studies may pave the way to the use of ß-glucosidase inhibitors for halitosis clinical management.

Identification and Characterization of Multidrug-Resistant Extended-Spectrum Beta-Lactamase-Producing Bacteria from Healthy and Diseased Dogs and Cats Admitted to a Veterinary Hospital in Brazil.

The objective of this study was to identify the main extended-spectrum beta-lactamase (ESBL)-producing bacteria and to detect the frequency of the major genes responsible to trigger this resistance in hospitalized animals. We collected 106 rectal swabs from cats (n = 25) and dogs (n = 81) to detect ESBL-producing isolates. ESBL-positive samples were submitted to the antimicrobial susceptibility test, and polymerase chain reaction was performed to detect TEM, SHV, and CTX-M genes from different groups. We observed that 44.34% of these samples (11 cats and 36 dogs) were positive for ESBL-producing bacteria. Thirteen animals (27.66%-seven cats and six dogs) were hospitalized for elective castration (healthy animals). Only a single animal was positive for ESBL-producing bacteria at hospital admission (the animal also showed an ESBL-positive isolate after leaving the hospital), whereas 11 were positive only at the hospital discharge. Of the 73 ESBL-producing isolates, 13 were isolated from cats (8 sick and 7 healthy) and 60 from dogs (53 sick and 7 healthy). Escherichia coli was the major ESBL-producing bacterium isolated (53.42%), followed by Pseudomonas aeruginosa (15.07%), Salmonella sp., and Proteus mirabilis (5.48% each one). Antimicrobial resistance profile of ESBL-producing isolates showed that 67 isolates (91.78%) were resistant to 3 or more antibiotic classes, while 13 of them (17.81%-2 healthy cats and 11 sick dogs) were resistant to all tested antimicrobial classes. The blaTEM gene exhibited the highest frequency in ESBL-producing isolates, followed by the blaCTX-M group 8/25, blaCTX-M group 1 and blaCTX-M group 9 genes. These results are useful to assess the predominance of ESBL-producing isolates recovered from dogs and in cats in Brazil. Consequently, we draw attention to these animals, as they can act as reservoirs for these microorganisms, which are the major pathogens of nosocomial infections worldwide.

Detection of the main multiresistant microorganisms in the environment of a teaching veterinary hospital in Brazil

ABSTRACT: Contamination of the veterinary hospital environment with multiresistant pathogens endangers not only hospitalized animals, but also the workplace safety of veterinarians and nurses, animal guardians and, when in case of a teaching hospital, veterinary students. The objective of this study was to map the main points of bacterial contamination of a veterinary teaching hospital in Brazil to identify multiresistant microorganisms and their antimicrobial resistance genes. Samples were collected from 39 different locations of a veterinary school hospital which comprised a pool according to each hospital environment. In certain environments, more than one pool has been collected. All samples were collected in quadruplicates for the selective isolation of the main multiresistant microorganisms: methicillin-resistant Staphylococcus (MRS), vancomycin resistant Enterococcus (VRE), cephalosporinases and/or extended-spectrum beta-lactamase-producing Gram-negative bacteria (ESBL) and Carbapenemase-producing (CP). After isolation and identification of isolates, multiplex-PCR reactions were performed to detect the main genes for each microorganism and antimicrobial susceptibility tests with the main antibiotics used for each bacterial group according to CLSI. Of the 39 veterinary teaching hospital sites collected, all (100%) had at least one of the microorganisms surveyed, and 17.95% (n=7) of the sites were able to isolate the four pathogens. From the 94 pools collected, it was possible to isolate MRS in 81.91% (n=77), VRE in 12.77% (n=12), cephalosporinases and/or ESBL in 62.77% (n=59) and CP in 24.47%. (n=23). Regarding MRS, the mecA gene was detected in all isolates. All isolated VREs were identified as Enterococcus faecalis and presented the vanA gene. Regarding cephalosporinases and/or ESBL, 89.83% (n=53) of the isolates presented the blaTEM gene, 57.63% (n=34) the blaOXA-1 gene, 37.29% (n=22) blaCTX-M gene from some group (1, 2, 9 ou 8/25) and 20.34% (n=12) the blaSHV gene. It was possible to identify the main microorganisms responsible for causing nosocomial infections in humans (VRE, MRS, ESBL and CP) in the veterinary hospital environment, suggesting a source of infection for professionals and students of veterinary medicine, placing a high risk for public health.

RESUMO: A contaminação do ambiente hospitalar veterinário com patógenos multirresistentes coloca em perigo não apenas os animais hospitalizados, mas também a segurança no local de trabalho de veterinários e enfermeiros, responsáveis por animais e, quando se tratar de um hospital de ensino, estudantes de veterinária. O objetivo deste estudo foi mapear os principais pontos de contaminação bacteriana de um hospital veterinário de ensino no Brasil, identificando microorganismos multirresistentes e seus genes de resistência antimicrobiana. As amostras foram coletadas em 39 locais diferentes de um hospital de escola veterinária, que compreendia um pool de acordo com o ambiente de cada hospital. Em certos ambientes, mais de um pool foi coletado. Todas as amostras foram coletadas em quadruplicados para o isolamento seletivo dos principais microorganismos multirresistentes: Staphylococcus resistente à meticilina (MRS), Enterococcus resistente à vancomicina (VRE), bactérias Gram-negativas produtoras de cefalosporinases e/ou beta-lactamase de espectro estendido (ESBL) e produtoras de carbapenemase (PC). Após o isolamento e identificação dos isolados, foram realizadas reações de PCR multiplex para detectar os principais genes de cada microorganismo e testes de susceptibilidade a antimicrobianos com os principais antibióticos utilizados para cada grupo bacteriano de acordo com o CLSI. Dos 39 locais do VCH coletados, todos (100%) possuíam pelo menos um dos microrganismos pesquisados e 17,95% (n=7) dos locais foram capazes de isolar os quatro patógenos. Dos 94 pools coletados, foi possível isolar MRS em 81,91% (n=77), VRE em 12,77% (n=12), ESBL em 62,77% (n=59) e carbapenemases em 24,47% (n=23). Em relação ao MRS, o gene mecA foi detectado em todos os isolados. Todos os VREs isolados foram identificados como Enterococcus faecalis e apresentaram o gene vanA. Em relação às cefalosporinases e/ou ESBL, 89,83% (n=53) dos isolados apresentaram o gene blaTEM, 57,63% (n=34) o gene blaOXA-1, 37,29% (n=22) o gene blaCTX-M de algum grupo e 20,34% (n=12) o gene blaSHV. Foi possível identificar os principais microrganismos responsáveis por causar infecções nosocomiais em humanos (VRE, MRS, ESBL e CP) no ambiente hospitalar veterinário, sugerindo uma fonte de infecção para profissionais e estudantes de medicina veterinária, colocando alto risco para a saúde pública.

Detection of the main multiresistant microorganisms in the environment of a teaching veterinary hospital in Brazil / Detecção dos principais microrganismos multirresistentes no ambiente de um hospital veterinário de ensino no Brasil

Contamination of the veterinary hospital environment with multiresistant pathogens endangers not only hospitalized animals, but also the workplace safety of veterinarians and nurses, animal guardians and, when in case of a teaching hospital, veterinary students. The objective of this study was to map the main points of bacterial contamination of a veterinary teaching hospital in Brazil to identify multiresistant microorganisms and their antimicrobial resistance genes. Samples were collected from 39 different locations of a veterinary school hospital which comprised a pool according to each hospital environment. In certain environments, more than one pool has been collected. All samples were collected in quadruplicates for the selective isolation of the main multiresistant microorganisms: methicillin-resistant Staphylococcus (MRS), vancomycin resistant Enterococcus (VRE), cephalosporinases and/or extended-spectrum beta-lactamase-producing Gram-negative bacteria (ESBL) and Carbapenemase-producing (CP). After isolation and identification of isolates, multiplex-PCR reactions were performed to detect the main genes for each microorganism and antimicrobial susceptibility tests with the main antibiotics used for each bacterial group according to CLSI. Of the 39 veterinary teaching hospital sites collected, all (100%) had at least one of the microorganisms surveyed, and 17.95% (n=7) of the sites were able to isolate the four pathogens. From the 94 pools collected, it was possible to isolate MRS in 81.91% (n=77), VRE in 12.77% (n=12), cephalosporinases and/or ESBL in 62.77% (n=59) and CP in 24.47%. (n=23). Regarding MRS, the mecA gene was detected in all isolates. All isolated VREs were identified as Enterococcus faecalis and presented the vanA gene. Regarding cephalosporinases and/or ESBL, 89.83% (n=53) of the isolates presented the blaTEM gene, 57.63% (n=34) the blaOXA-1 gene, 37.29% (n=22) blaCTX-M gene from some group (1, 2, 9 ou 8/25) and 20.34% (n=12) the blaSHV gene. It was possible to identify the main microorganisms responsible for causing nosocomial infections in humans (VRE, MRS, ESBL and CP) in the veterinary hospital environment, suggesting a source of infection for professionals and students of veterinary medicine, placing a high risk for public health.(AU)

A contaminação do ambiente hospitalar veterinário com patógenos multirresistentes coloca em perigo não apenas os animais hospitalizados, mas também a segurança no local de trabalho de veterinários e enfermeiros, responsáveis por animais e, quando se tratar de um hospital de ensino, estudantes de veterinária. O objetivo deste estudo foi mapear os principais pontos de contaminação bacteriana de um hospital veterinário de ensino no Brasil, identificando microorganismos multirresistentes e seus genes de resistência antimicrobiana. As amostras foram coletadas em 39 locais diferentes de um hospital de escola veterinária, que compreendia um pool de acordo com o ambiente de cada hospital. Em certos ambientes, mais de um pool foi coletado. Todas as amostras foram coletadas em quadruplicados para o isolamento seletivo dos principais microorganismos multirresistentes: Staphylococcus resistente à meticilina (MRS), Enterococcus resistente à vancomicina (VRE), bactérias Gram-negativas produtoras de cefalosporinases e/ou beta-lactamase de espectro estendido (ESBL) e produtoras de carbapenemase (PC). Após o isolamento e identificação dos isolados, foram realizadas reações de PCR multiplex para detectar os principais genes de cada microorganismo e testes de susceptibilidade a antimicrobianos com os principais antibióticos utilizados para cada grupo bacteriano de acordo com o CLSI. Dos 39 locais do VCH coletados, todos (100%) possuíam pelo menos um dos microrganismos pesquisados e 17,95% (n=7) dos locais foram capazes de isolar os quatro patógenos. Dos 94 pools coletados, foi possível isolar MRS em 81,91% (n=77), VRE em 12,77% (n=12), ESBL em 62,77% (n=59) e carbapenemases em 24,47% (n=23). Em relação ao MRS, o gene mecA foi detectado em todos os isolados. Todos os VREs isolados foram identificados como Enterococcus faecalis e apresentaram o gene vanA. Em relação às cefalosporinases e/ou ESBL, 89,83% (n=53) dos isolados apresentaram o gene blaTEM, 57,63% (n=34) o gene blaOXA-1, 37,29% (n=22) o gene blaCTX-M de algum grupo e 20,34% (n=12) o gene blaSHV. Foi possível identificar os principais microrganismos responsáveis por causar infecções nosocomiais em humanos (VRE, MRS, ESBL e CP) no ambiente hospitalar veterinário, sugerindo uma fonte de infecção para profissionais e estudantes de medicina veterinária, colocando alto risco para a saúde pública.(AU)

Enterotoxigenic potential of Staphylococcus spp. isolates recovered from raw milk and artisanal cheese.

In this work, we investigated the phenotypic profile of Staphylococcus spp. isolates recovered from raw milk and artisanal cheese, and their enterotoxigenic potential through the detection of classical enterotoxin genes (sea, seb, sec, sed and see). A total of 104 isolates (58 coagulase-positive Staphylococcus - CoPS; and 46 coagulase-negative Staphylococcus- CoNS) were used, of which 33 were retrieved from raw milk and 71 from artisanal cheese produced in the Serrana region of Santa Catarina. Identification of CoPS was conducted via biochemical tests. Detection of the genes sea, seb, sec, sed, and see was carried out by multiplex PCR technique. Among the 58 CoPS analyzed, 64% were identified as S. aureus, 22% as S. scheiferi coagulans, 12% as S. hyicus and as 2% S. intermedius. In the present study was noted that 40% of CoPS isolates retrieved from milk harbored seb gene, while only one from artisanal cheese was positive for gene sea. In this study all CoNS samples investigated were negative for enterotoxins genes. The enterotoxigenic potential of CoPS, is an issue of great importance for public health. For that reason, it is necessary that cheese factories strictly follow the safety processes involved in manufacturing.

ASPIRATION BRONCHOPNEUMONIA BY ACINETOBACTER BAUMANNII IN A WILDLIFE EUROPEAN HARE (LEPUS EUROPAEUS) IN BRAZIL.

Acinetobacter baumannii is a major cause of illness in hospitalized patients and the most important and common pathogen in nosocomial outbreaks worldwide. In animals, A. baumannii has been associated with respiratory infections in a group of minks, leading to pneumonia and acute mortality. This report documents a case of aspiration bronchopneumonia in a wild European hare caused by A. baumannii. A free-ranging, adult male European hare was submitted to necropsy after acute trauma due to being hit by a car. Its lungs showed consolidation with abscess in the middle and cranial lobes. Histopathologic evaluation revealed liquefactive necrosis associated with neutrophilic infiltration, cellular debris, plant material, and bacterial myriads surrounded by moderate neutrophils, macrophages, multinucleated giant cells, lymphocytes, and plasma cell inflammation. Acinetobacter baumannii was isolated from lung tissue.

Surgical treatment of postoperative laparostomy and pyloroduodenostomy-Case report.

INTRODUCTION: Postoperative duodenal-cutaneous fistula represents a rare and very complex problem. In most cases operative management becomes necessary, but only after local and systemic stabilization and sepsis control. CASE PRESENTATION: A 39-year-old man was admitted for surgical management of laparostomy and pyloro-duodenostomy of the first (DI) and second (DII) duodenal segments with one year of evolution, as a complication of several surgical interventions. The patient had been previously submitted to surgical interventions in another institution for: 1- lower gastrointestinal haemorrhage: treated with total colectomy; 2- upper gastrointestinal haemorrhage: performed a pyloroduodenotomy and pyloroplasty; 3- evisceration: abdominal wall closure; 4- biliary peritonitis due to pyloroplasty dehiscence: submitted to laparotomy with placement of a gastrostomy tube and pyloroduodenostomy tube; 5- intestinal haemorrhage through the pyloroduodenostomy tube: inconclusive exploratory laparotomy plus laparostomy; 6- gastrointestinal haemorrhage and shock: submitted to jejunal segmental resection (haemorrhagic mucous nodule); 7- several complications related to drainage, fistulae and celiostomy. DISCUSSION: After initial medical treatment for local and systemic stabilization during four months, the following surgical procedures were performed: antrectomy; duodenectomy of DI and the suprapapillary part of DII; T-L gastrojejunostomy; duodenojejunostomy (DII and DIII) L-L at 40 cm of the gastrojejunal anastomosis; T-L jejunojejunostomy; abdominoplasty with a mesh and fibrin glue application; primary cutaneous closure. A multitubular drain was positioned near the duodeno-jejunal anastomosis and a suction drain was positioned in the subcutaneous space. CONCLUSION: The patient was discharged at the 60th postoperative day, asymptomatic and with a weight gain of 10 kg.

Low occurrence of pathogenic Yersinia enterocolitica in pig tonsils at slaughter in Southern Brazil.

Yersinia enterocolitica is a foodborne pathogen and pigs are the main reservoir of it in their tonsils. As Brazil is a large producer and exporter of pork meat and information regarding this pathogen is still quite scarce, this study aimed at evaluating the direct detection of Y. enterocolitica followed by pathogenic Y. enterocolitica (PYE) determination in tonsils of slaughtered pigs. For this purpose, 400 pig tonsils were collected from 15 farms in four federally certified slaughterhouses in Southern Brazil. Initially, samples were screened using conventional PCR targeting of the 16sRNA gene, followed by multiplex PCR (mPCR) in order to detect three virulence genes (ail, yadA, and virF) and quantitative real-time PCR (qPCR) for the detection of the ail gene. One hundred and one (25.2%) of the samples tested positive for the 16sRNA gene. However, a PYE was detected in one out of the 101 Y. enterocolitica positive samples. The three virulence genes were determined by mPCR and confirmed by partial DNA sequencing. Thus, a significant occurrence of Y. enterocolitica was observed in pig tonsils from federally inspected slaughterhouses in Brazil, although the presence of pathogenic strains was quite low.

Heterogeneity determination of bovine leukemia virus genome in Santa Catarina state, Brazil / Determinação da heterogeneidade do genoma do vírus da leucose enzoótica bovina no estado de Santa Catarina, Brasil

Bovine leukemia virus (BLV) is a member of Retroviridae family, genus Deltaretrovirus, and the main viral agent responsible for economic loses in dairy herds. Some studies have been carried out about BLV genotypes, and at least seven genotypes were found out in samples of different regions of the world. The objective of this study was to identify BLV samples from seropositive dairy cattle in Santa Catarina state, Brazil, using molecular techniques. Blood samples were collected (454) from dairy cattle from 31 different farms, and serology using agar gel immunodiffusion test (AGID) was performed. After that, 191 seropositive samples were submitted to DNA extraction, and in 77 samples the polymerase chain reaction (PCR) for amplification of a 440 bp fragment of the env gene was performed. Nineteen DNA samples were subjected to restriction fragment length polymorphism (RFLP) analysis by digestion of the PCR fragment by five restriction endonucleases - BamHI, HaeIII, Tru9I, TaqI, and MwoI. It was found 42% seropositive animals (191/454) and 68% positives of the farms (21/31). The PCR showed 80.5% (62/77) of animals positive. The RFLP analysis identified five different genotypes dispersed by Santa Catarina state, with the highest prevalence for genotype X (47.4%). Overall, our results identified the viral genotypes present in dairy cattle and the prevalence of new variants in representative farms from Santa Catarina state.(AU)

O bovine leukemia virus (BLV) é um membro da família Retroviridae, gênero Deltaretrovirus, e o principal agente viral causador de perdas econômicas em rebanhos leiteiros. Diversos estudos têm sido feitos sobre os genótipos de BLV, e foram encontrados pelo menos sete em amostras de diferentes partes do mundo. O objetivo deste estudo foi realizar a caracterização molecular de amostras de BLV de bovinos leiteiros soropositivos no estado de Santa Catarina. Foram coletadas 454 amostras de sangue de bovinos de 31 propriedades, e fez-se inicialmente a sorologia por meio do teste de imunodifusão em gel de ágar. Após a sorologia, 191 amostras soropositivas foram então submetidas à extração de DNA, e em 77 amostras se realizou a reação da polimerase em cadeia (PCR), para a amplificação de um fragmento de 440 pb do gene env. Dezenove amostras foram submetidas à análise do polimorfismo dos fragmentos de restrição por digestão do fragmento da PCR por cinco enzimas de restrição: BamHI, HaeIII, Tru9I, TaqI e MwoI. Os resultados obtidos na sorologia apontaram 42% de animais soropositivos (191/454) e 68% de propriedades positivas (21/31). Na PCR, 80,52% (62/77) dos animais apresentaram-se positivos. A análise do polimorfismo dos fragmentos de restrição identificou cinco genótipos circulantes no estado, e a maior prevalência foi observada no genótipo X (47,4%). Este estudo permite-nos conhecer alguns dos genótipos virais presentes em bovinos leiteiros do estado de Santa Catarina, bem como identificar a existência de novas variantes e sua prevalência atual, e os resultados são úteis para futuros estudos epidemiológicos.(AU)

Brucella abortus RB51 in milk of vaccinated adult cattle.

The aim of this study was to evaluate the shedding of Brucella abortus in the milk of cows vaccinated with a full dose of RB51 during lactation. Eighteen cows, nine previously vaccinated with S19 as calves and nine non-vaccinated, were immunized subcutaneously with 1.3×10(10)CFU of B. abortus RB51, 30-60days after parturition. Milk samples from all animals were collected daily until day 7, and at weekly interval for the next 9 weeks after vaccination. To evaluate the shedding of B. abortus, milk samples were submitted for culture and PCR. No B. abortus was isolated from any sample tested. Only one sample, collected on first day after vaccination from a cow previously vaccinated, was faintly positive in the PCR. In conclusion, the public health hazard associated with milk consumption from cows vaccinated with RB51 in post-partum is very low, despite vaccination with the full dose and regardless of previous S19 vaccination.