RESUMO O presente trabalho estudou, através de modelagem matemática, a dinâmica da infecção por Mycobacterium avium em uma população suína. Para descrever a dinâmica de transmissão de M. avium em granjas de suínos, foi elaborado um modelo de compartimentos. Como esse modelo visava estimar a relação entre taxa de infecção e proporção de suínos que apresentam lesões, detectadas no abate aos 150 dias de idade, resultantes de infecção por M. avium, considerou-se que a dinâmica da doença poderia ser descrita por dois compartimentos de animais: suscetíveis à infecção, e infectados que poderiam vir a apresentar lesões. Através dos resultados obtidos nas simulações realizadas, observou-se que para uma dada força de infecção, a proporção de infectados no final da fase de creche (aqueles que apresentariam lesões no abate) se mantém constante ao longo do tempo, não estando sujeita a variações de caráter sazonal. A simulação matemática da doença, considerando a transmissão horizontal como mecanismo principal da ocorrência de condenações em matadouro por linfadenite granulomatosa é inconsistente com o que se observa na população. Portanto, o componente ambiental tem papel preponderante na dinâmica das infecções micobacterianas dos suínos produzidos no Brasil.

ABSTRACT The present work studied, by mathematical modeling, the Mycobacterium avium infection dynamics in a swine population. In order to describe M. avium transmission dynamics, a compartmental model was elaborated. Since the model was aimed to estimate the relationship between infection rate and the rate of swine that presented lesions resulting from M. avium infection, detected at slaughter of 150-day-old animals, it was considered that the disease dynamics could be described by two compartments of animals: susceptible to the infection, and infected that could present lesions. The simulation’s results showed that for a certain force of infection, the proportion of infected animals at the end of the nursery stage (those that would present lesions at the slaughter) remains constant over time, not being subject to seasonal changes. The mathematical simulation of the disease, considering horizontal transmission as the major means of condemnation at slaughter due to granulomatous lymphadenitis, is inconsistent with what is observed within the population. Therefore, the environmental component plays a major/preponderant role in the mycobacterial infection dynamics of swine raised on farms in Brazil.

In order to develop a new experimental animal model of infection with Mycobacterium chelonae in keratomileusis, we conducted a double-blind prospective study on 24 adult male New Zealand rabbits. One eye of each rabbit was submitted to automatic lamellar keratotomy with the automatic corneal shaper under general anesthesia. Eyes were immunosuppressed by a single local injection of methyl prednisolone. Twelve animals were inoculated into the keratomileusis interface with 1 µl of 10(6) heat-inactivated bacteria (heat-inactivated inoculum controls) and 12 with 1 µl of 10(6) live bacteria. Trimethoprim drops (0.1%, w/v) were used as prophylaxis for the surgical procedure every 4 h (50 µl, qid). Animals were examined by 2 observers under a slit lamp on the 1st, 3rd, 5th, 7th, 11th, 16th, and 23rd postoperative days. Slit lamp photographs were taken to document clinical signs. Animals were sacrificed when corneal disease was detected and corneal samples were taken for microbiological analysis. Eleven of 12 experimental rabbits developed corneal disease, and M. chelonae could be isolated from nine rabbits. Eleven of the 12 controls receiving a heat-inactivated inoculum did not develop corneal disease. M. chelonae was not isolated from any of the control rabbits receiving a heat-inactivated inoculum, or from the healthy cornea of control rabbits. Corneal infection by M. chelonae was successfully induced in rabbits submitted to keratomileusis. To our knowledge, this is the first animal model of M. chelonae infection following corneal flaps for refractive surgery to be described in the literature and can be used for the analysis of therapeutic responses.

ABSTRACT A study was performed in nine swine slaughterhouses of the South of Brazil, responsible for almost 40% of total swine slaughtered in this area, to verify if the condemnations of carcasses by granulomatous lymphadenitis were seasonal during the period of January 1996 to December 1999. Therefore, forms were filled with monthly information about total number of swine slaughtered and occurrence of carcasses condemnations for granulomatous lymphadenitis at the slaughter. The higher levels of condemnations occurred from June to October (p = 0.034) there and was also verified a cyclic variation each two years. The implications and possible reasons of these findings were discussed regarding the improvement of the control program. Brazil 40 area 199 1999 Therefore slaughter p 0.034 0034 0 034 years program 4 19 0.03 003 03 1 0.0 00 0.

RESUMO Foi realizado um estudo em 9 abatedouros de suínos da Região Sul do Brasil, responsáveis por cerca de 40% do total de abate na Região, para verificar se a condenação de carcaças por linfadenites granulomatosas apresentou comportamento sazonal durante o período de janeiro de 1996 a dezembro de 1999. Para tanto, foram preenchidas planilhas com informações mensais a respeito do número total de suínos abatidos e da ocorrência dessas condenações. Os resultados mostraram que essas condenações apresentaram picos de ocorrência nos períodos de junho a outubro (p = 0,034) e também foi verificada uma variação cíclica com picos a cada 2 anos. As implicações e razões destas constatações foram discutidas com vistas à estruturação de um programa de controle racional. Brasil 40 199 1999 tanto p 0,034 0034 0 034 anos racional 4 19 0,03 003 03 1 0,0 00 0,

ABSTRACT A study was performed in nine swine slaughterhouses of the South of Brazil, responsible for around 40% of total swine slaughtered in this area, with the aim to estimate the economic impact of micobacteriosis for swine producers. The period studied was from January 1997 to December 1999. To do so, forms were filled with mensal informations about total number of swine slaughtered and occurrence of carcasses condemnations for granulomatous lymphadenitis at the slaughter; other form gave informations about the discounts received by producers depending on the carcasses destiny and about the average value of bonus received for carcass typing. The economic impact was 1,5 million of reais in 1997, 4,27 in 1998 and 6,94 in 1999. Each 0.1% of increment in the prevalence of swine tuberculosis condemnations represented a loss of 201 thousand of reais in 1997, 531 in 1998 and 834 in 1999. The implications and possible reasons of these findings were discussed regarding the elaboration of a rational control program. Brazil 40 area 199 1999 so slaughter typing 15 1 5 1, 427 4 27 4,2 694 6 94 6,9 01 0 0.1 20 53 83 program 19 42 2 4, 69 9 6, 0. 8

RESUMO Foi realizado um estudo em nove abatedouros de suínos da Região Sul do Brasil, responsáveis por cerca de 40% do total de abate na Região, com o objetivo de se estimar o impacto econômico das micobacterioses para os produtores. O período estudado foi de janeiro de 1997 a dezembro de 1999. Para tanto, foram preenchidas planilhas com informações mensais a respeito do número total de suínos abatidos e ocorrência de condenação por linfadenite granulomatosa na ocasião do abate; outra planilha forneceu dados sobre os descontos impostos aos produtores dependendo do destino da carcaça condenada e valor médio da bonificação recebida pela sua tipificação. O impacto econômico foi de 1,5 milhão de reais em 1997; 4,3 milhões em 1998 e 6,9 em 1999. Cada 0,1% de incremento na prevalência das condenações por linfadenite granulomatosa representou um prejuízo de 201 mil reais em 1997, 531 em 1998 e 834 em 1999. As implicações e razões destas constatações foram discutidas com vistas à estruturação de um programa de controle racional. Brasil 40 199 1999 tanto tipificação 15 1 5 1, 43 4 3 4, 69 6 9 6, 01 0 0,1 20 53 83 racional 19 0, 2 8

ABSTRACT IDENTIFICATION OF RISK FACTORS FOR SWINE MYCOBACTERIOSIS IN THE SOUTH OF BRAZIL. A case control study was performed to identify the risk factors associated with the occurrence of swine mycobacteriosis in the South Region of Brazil. Thirty three swine herds with more than two tuberculous pigs at slaughter and 66 herds with no evidence of infection were selected. Two control herds were chosen in a maximum of a 5 km ratio distance from each case. All selected herds were scrutinized through a questionnaire based on literature data and local characteristics. The results showed that an increase risk of infection is associated with: herds with ≥25 sows (OR= 4.2; 1.2≥IC95%≤14.5), nursery without partial or completely lath floor (OR=3.6; 1.05≥IC95%≤12.7), and bad quality of hygiene in the time of inquest (OR=2.9; 0.99≥IC95%≤8.3). The implementation of hygiene and disinfection program was suggested as strategy of control, obviously observing the economic aspects and the resistance of the mycobacteria to the environment and livestock disinfectants. BRAZIL Brazil 6 characteristics 25 ≥2 OR= OR (OR 4.2 42 4 2 1.2≥IC95%≤14.5, 12IC95145 IC 1.2≥IC95%≤14.5 , 1 IC95 14 1.2≥IC95%≤14.5) OR=3.6 OR36 3 (OR=3.6 1.05≥IC95%≤12.7, 105IC95127 1.05≥IC95%≤12.7 05 12 7 1.05≥IC95%≤12.7) OR=2.9 OR29 9 (OR=2.9 0.99≥IC95%≤8.3. 099IC9583 0.99≥IC95%≤8.3 . 0 99 8 0.99≥IC95%≤8.3) disinfectants ≥ 4. 12IC9514 1.2≥IC95%≤14. IC9 OR=3. OR3 (OR=3. 105IC9512 1.05≥IC95%≤12. OR=2. OR2 (OR=2. 099IC958 0.99≥IC95%≤8. 12IC951 1.2≥IC95%≤14 OR=3 (OR=3 105IC951 1.05≥IC95%≤12 OR=2 (OR=2 099IC95 0.99≥IC95%≤8 12IC95 1.2≥IC95%≤1 105IC95 1.05≥IC95%≤1 099IC9 0.99≥IC95%≤ 12IC9 1.2≥IC95%≤ 105IC9 1.05≥IC95%≤ 099IC 0.99≥IC95% 12IC 1.2≥IC95% 105IC 1.05≥IC95% 0.99≥IC95 1.2≥IC95 1.05≥IC95 0.99≥IC9 1.2≥IC9 1.05≥IC9 0.99≥IC 1.2≥IC 1.05≥IC

RESUMO Foi realizado um estudo caso-controle para identificação de fatores de risco associados à ocorrência de micobacterioses em suínos, na região sul do Brasil. Todas as granjas selecionadas foram do tipo ciclo completo. Foram consideradas casos aquelas que apresentaram dois ou mais animais condenados por “tuberculose”, confirmadas por exames histológicos e/ou microbiológicos. Em raio inferior a 5 km de cada caso foram selecionados dois controles com histórico de não condenação por no mínimo dois anos. Nas 33 granjas caso e nas 66 controles foi aplicado um questionário elaborado com base em dados de literatura e na realidade local, procurando investigar variáveis que poderiam estar associadas a tais infecções. O modelo final da regressão logística apontou três variáveis associadas a infecções por micobactérias: dimensão do rebanho ≥25 matrizes (OR= 4,2; 1,2≥IC95%≤14,5), não ter piso ripado ou parcialmente ripado nas instalações de creche (OR=3,6; 1,05≥IC95%≤12,7); má qualidade da higiene da creche por ocasião da visita; (OR=2,9; 0,99≥IC95%≤8,3). Foi sugerido como estratégia de controle a adoção de programa de higiene e desinfecção, com especial atenção para a fase de creche, naturalmente bem planejados do ponto de vista econômico e respeitando a singular resistência das micobactérias ao meio ambiente e a desinfetantes de uso pecuário. casocontrole suínos Brasil completo tuberculose, tuberculose , “tuberculose” eou microbiológicos anos 3 6 local 25 ≥2 OR= OR (OR 4,2 42 4 2 1,2≥IC95%≤14,5, 12IC95145 IC 1,2≥IC95%≤14,5 1 IC95 14 1,2≥IC95%≤14,5) OR=3,6 OR36 (OR=3,6 1,05≥IC95%≤12,7 105IC95127 05 12 7 1,05≥IC95%≤12,7) visita OR=2,9 OR29 9 (OR=2,9 0,99≥IC95%≤8,3. 099IC9583 0,99≥IC95%≤8,3 . 0 99 8 0,99≥IC95%≤8,3) desinfecção pecuário “tuberculose ≥ 4, 12IC9514 1,2≥IC95%≤14, IC9 OR=3, OR3 (OR=3, 1,05≥IC95%≤12, 105IC9512 OR=2, OR2 (OR=2, 099IC958 0,99≥IC95%≤8, 12IC951 1,2≥IC95%≤14 OR=3 (OR=3 1,05≥IC95%≤12 105IC951 OR=2 (OR=2 099IC95 0,99≥IC95%≤8 12IC95 1,2≥IC95%≤1 1,05≥IC95%≤1 105IC95 099IC9 0,99≥IC95%≤ 12IC9 1,2≥IC95%≤ 1,05≥IC95%≤ 105IC9 099IC 0,99≥IC95% 12IC 1,2≥IC95% 1,05≥IC95% 105IC 0,99≥IC95 1,2≥IC95 1,05≥IC95 0,99≥IC9 1,2≥IC9 1,05≥IC9 0,99≥IC 1,2≥IC 1,05≥IC

The genome of Mycobacterium tuberculosis H37Rv contains three contiguous genes (plc-a, plc-b and plc-c) which are similar to the Pseudomonas aeruginosa phospholipase C (PLC) genes. Expression of mycobacterial PLC-a and PLC-b in E. coli and M. smegmatis has been reported, whereas expression of the native proteins in M. tuberculosis H37Rv has not been demonstrated. The objective of the present study was to demonstrate that native PLC-a is expressed in M. tuberculosis H37Rv. Sera from mice immunized with recombinant PLC-a expressed in E. coli were used in immunoblots to evaluate PLC-a expression. The immune serum recognized a 49-kDa protein in immunoblots against M. tuberculosis extracts. No bands were visible in M. tuberculosis culture supernatants or extracts from M. avium, M. bovis and M. smegmatis. A 550-bp DNA fragment upstream of plc-a was cloned in the pJEM12 vector and the existence of a functional promoter was evaluated by detection of ß-galactosidase activity. ß-Galactosidase activity was detected in M. smegmatis transformed with recombinant pJEM12 grown in vitro and inside macrophages. The putative promoter was active both in vitro and in vivo, suggesting that expression is constitutive. In conclusion, expression of non-secreted native PLC-a was demonstrated in M. tuberculosis.