Occurrence of F42 colonization factor in Escherichia coli strains isolated from piglets with diarrhea

The objective of this study was to determine the presence of the colonization factor F42 in 168 strains of Escherichia coli isolated from diarrheic stools of newborn piglets. The presence of F42 in 12 (7.1%) strains was detected with the agglutination test. Through the Polymerase Chain Reaction (PCR) of F42 positive strains, gene encoding enterotoxins (ST-I, ST-II, LT-I and LT-II) were detected. The finding of ST-I/ST-II genes in 50% of the strains, ST-I (16%) and ST-II (25%) indicates a strong association of FC F42 with heat-stable enterotoxins (91%). In contrast, the thermolabile enterotoxin (LT-I and LT-II) genes were not detected. Serogroups of F42 positive strains were determined, serogroup O8 being the most prevalent (41,7%). Other serogroups, as there are O9, O11, O18, O32, O35, O98 and O101, were also identified. Thus, FC F42 was confirmed as an additional factor of virulence in the pathogenesis of porcine colibacillosis.

Seroprevalence of Leptospira spp in cattle from Monte Negro municipality, western Amazon

The prevalence of anti-Leptospira spp antibodies was investigated in 2,109 female cattle from 86 herds of Monte Negro municipality, Rondônia, Brazil. Sera samples were evaluated by Microscopic Agglutination Test against 24 leptospira serovars. Titers =100 for at least one of 24 leptospira serovars were detected in 1,114 cows (52.8%) from 82 (95.3%) herds. The adjusted overall prevalence for Monte Negro municipality was 53.9% (49-58.7%; CI: 95%). The most prevalent serovars were Hardjo (14.5%), Wolffi (12.3%), Shermani (10.8%), Patoc (7.9%), and Hebdomadis (6.1%). Other serovars worldwidely reported like Bratislava, Pomona and Grippotyphosa were detected in low levels.

Matched case-control study evaluating the frequency of the main agents associated with neonatal diarrhea in piglets

A case-control study was carried out in litters of 1 to 7-day-old piglets to identify the main infectious agents involved with neonatal diarrhea in pigs. Fecal samples (n=276) from piglets were collected on pig farms in the State of Rio Grande do Sul, Brazil, from May to September 2007. Litters with diarrhea were considered cases (n=129) and normal litters (n=147) controls. The samples were examined by latex agglutination test, PAGE, conventional isolating techniques, ELISA, PCR, and microscopic methods in order to detect rotavirus, bacterial pathogens (Escherichia coli, Clostridium perfringens type A and C, and Clostridium difficile), and parasites (Coccidian and Cryptosporidium spp.). Outbreaks of diarrhea were not observed during sampling. At least one agent was detected in fecal samples on 25 out of 28 farms (89.3%) and in 16 farms (57.1%) more than one agent was found. The main agents diagnosed were Coccidia (42.86%) and rotavirus (39.29%). The main agents identified in litters with diarrhea were Clostridium difficile (10.6%), Clostridium perfringens type A (8.8%) and rotavirus (7.5%); in control litters, Clostridium difficile (16.6%) and Coccidian (8.5%). Beta hemolytic Escherichia coli and Clostridium perfringens type C were not detected. When compared with controls, no agent was significantly associated with diarrhea in case litters. These findings stress the need for caution in the interpretation of laboratorial diagnosis of mild diarrhea in neonatal pigs, as the sole detection of an agent does not necessarily indicate that it is the cause of the problem.

Efficacy of bacterin-, outer membrane protein- and fimbriae extract-based vaccines for the control of Salmonella Enteritidis experimental infection in chickens

The efficacy of three vaccines was evaluated in chickens for the control of experimental infection with Salmonella Enteritidis (SE) phage type 4. The vaccines were produced with bacterin, outer membrane proteins (OMP) and fimbriae crude extract (FE). The chickens were vaccinated intramuscularly with two doses of each vaccine at 12 and 15 weeks of age. The chickens were then orally challenged with 10(9) CFU/chicken Salmonella Enteritidis phage type 4 at 18 weeks of age. Fecal swabs were performed for the recovery of shedding SE, and SE was recovered from the liver and spleen. Additionally, antibody titers were measured in the serum by micro-agglutination test. The results indicated that the vaccine produced with bacterin yielded better results and resulted in reduction of fecal shedding and organ invasion by SE after oral challenge, although no vaccine was 100% effective for the control of SE experimental infection.

Serosurvey of Leptospirainterrogans, Brucellaabortus and Chlamydophilaabortus infection in free-ranging giant anteaters (Myrmecophaga tridactyla) from Brazil

Abstract:A serological survey for antibodies against Leptospira interrogans, Brucella abortus, and Chlamydophila abortus was conducted in 21 clinically healthy, free-ranging giant ant- eaters (Myrmecophaga tridactyla) from Parque Nacional das Emas (Goiás State, Brazil; n=6), Parque Nacional da Serra da Canastra (Minas Gerais State, Brazil; n=9), and RPPN SESC Pantanal (Mato Grosso State, Brazil; n=6) between July 2001 and September 2006. Sera were screened for antibodies against 22 serovars of Leptospira interrogans with a microscopic agglutination test. Twelve tested positive for L. interrogansserovars sentot (n=5 in PN Emas, n=2 in PN Serra da Canastra), butembo (n=2 in PN Serra da Canastra), autumnalis, bataviae, and shermani/icterohaemorrhagiae(n=1 each in SESC Pantanal)One adult female tested positive for B. abortus with the buffered plate antigen test. All sera were negative for C. abortususing the complement fixation text. This is the first report of pathogens that may interfere with the reproduction and population dynamics of free-ranging giant anteaters.

Anti-cyclic citrullinated peptide antibodies and rheumatoid factor in leprosy patients with articular involvement

The objective of the present research was to evaluate the usefulness of anti-cyclic citrullinated peptide (anti-CCP) antibodies and the IgM rheumatoid factor (IgM RF) test for the differential diagnosis of leprosy with articular involvement and rheumatoid arthritis (RA). Anti-CCP antibodies and IgM RF were measured in the sera of 158 leprosy patients (76 with and 82 without articular involvement), 69 RA patients and 89 healthy controls. Leprosy diagnosis was performed according to Ridley and Jopling classification criteria and clinical and demographic characteristics of leprosy patients were collected by a standard questionnaire. Leprosy patients with any concomitant rheumatic disease were excluded. Serum samples were obtained from all participants and frozen at _20°C. Measurement of anti-CCP antibodies and IgM RF were performed by ELISA, using a commercial second-generation kit, and the latex agglutination test, respectively. Anti-CCP antibodies and IgM RF were detected in low frequencies (2.6 and 1.3%, respectively) in leprosy patients and were not associated with articular involvement. Among healthy individuals both anti-CCP antibodies and IgM RF were each detected in 3.4% of the subjects. In contrast, in the RA group, anti-CCP antibodies were present in 81.2% and IgM RF in 62.3%. In the present study, both anti-CCP antibodies and IgM RF showed good positive predictive value for RA, helping to discriminate between RA and leprosy patients with articular involvement. However, anti-CCP antibodies were more specific for RA diagnosis in the population under study.

Immune strategies using single-component LipL32 and multi-component recombinant LipL32-41-OmpL1 vaccines against leptospira

Genes encoding lipoproteins LipL32, LipL41 and the outer-membrane protein OmpL1 of leptospira were recombined and cloned into a pVAX1 plasmid. BALB/c mice were immunized with LipL32 and recombined LipL32-41-OmpL1 using DNA-DNA, DNA-protein and protein-protein strategies, respectively. Prime immunization was on day 1, boost immunizations were on day 11 and day 21. Sera were collected from each mouse on day 35 for antibody, cytokine detection and microscopic agglutination test while spleen cells were collected for splenocyte proliferation assay. All experimental groups (N = 10 mice per group) showed statistically significant increases in antigen-specific antibodies, in cytokines IL-4 and IL-10, as well as in the microscopic agglutination test and splenocyte proliferation compared with the pVAX1 control group. The groups receiving the recombined LipL32-41-OmpL1 vaccine induced anti-LipL41 and anti-OmpL1 antibodies and yielded better splenocyte proliferation values than the groups receiving LipL32. DNA prime and protein boost immune strategies stimulated more antibodies than a DNA-DNA immune strategy and yielded greater cytokine and splenocyte proliferation than a protein-protein immune strategy. It is clear from these results that recombination of protective antigen genes lipL32, lipL41, and ompL1 and a DNA-protein immune strategy resulted in better immune responses against leptospira than single-component, LipL32, or single DNA or protein immunization.

Detection of enterotoxins produced by B. cereus through PCR analysis of ground and roasted coffee samples in Rio de Janeiro, Brazil

Coffee is one of the most appreciated drinks in the world. Coffee ground is obtained from the fruit of a small plant that belongs to the genus Coffea. Coffea arabica and Coffea canephora robusta are the two most commercially important species. They are more commonly known as arabica and robusta, respectively. Two-thirds of Coffea arabica plants are grown in South and Central America, and Eastern Africa - the place of origin for this coffee species. Contamination by microorganisms has been a major matter affecting coffee quality in Brazil, mainly due to the harvesting method adopted. Brazilian harvests are based on fruits collected from the ground mixed with those that fall on collection cloths. As the Bacillus cereus bacterium frequently uses the soil as its environmental reservoir, it is easily capable of becoming a contaminant. This study aimed to evaluate the contamination and potential of B. cereus enterotoxin genes encoding the HBL and NHE complexes, which were observed in strains of ground and roasted coffee samples sold in Rio de Janeiro. The PCR (Polymerase Chain Reaction) results revealed high potential of enterotoxin production in the samples. The method described by Speck (1984) was used for the isolation of contaminants. The investigation of the potential production of enterotoxins through isolates of the microorganism was performed using the B. cereus enterotoxin Reverse Passive Latex Agglutination test-kit (BCET-RPLA, Oxoid), according to the manufacturer's instructions. The potential of enterotoxin production was investigated using polymerase chain reaction (PCR) methods for hblA, hblD and hblC genes (encoding hemolysin HBL) and for nheA, nheB and nheC genes (encoding non-hemolytic enterotoxin - NHE). Of all the 17 strains, 100% were positive for at least 1 enterotoxin gene; 52.9% (9/17) were positive for the 3 genes encoding the HBL complex; 35.3% (6/17) were positive for the three NHE encoding genes; and 29.4% (5/17) were positive for all enterotoxic genes.

Teollisuusjätteen käyttö kiinteytys-stabiloinnin sideaineena ja haitta-aineiden stabiloinnin tehostamisessa

Tässä kanditaatintyössä selvitettiin kuinka erilaiset sideaineseokset soveltuvat raskas-metallien sitomiseen 28 vuorokautta stabiloiduissa näytteistä. Työssä oletettiin teollisten jätefraktioiden käytön tehostavan eräiden metallien, kuten kupari ja sinkki, immobilisointia lievästi pilaantuneista maa-aineksista. Kokeellisessa osassa stabiloitiin Kokkolan satamasta ruopattua sedimenttiä, jonka sinkkipitoisuudet olivat ylittäneet saastuneen sedimentin ohjearvon (≥400 mg/kg). Sedimenttiin lisättiin eri sideaineseoksia ja näytteiden annettiin stabiloitua 28 vuorokautta, minkä jälkeen niistä testattiin liukenevat raskasmetallit muokatulla ravistelutestillä. Eri sideaineseoksilla saatuja tuloksia verrattiin pelkän yleissementin käyttöön. Lisäksi erillisistä näytteistä otettiin pyyhkäisyelektronimikros-koopilla (SEM) kuvia havainnollistamaan stabiloitumista. Näissä näytteissä käytettiin samoja sideaineita kuin tehdyissä kokeissa. Liukoisuustestien tuloksista voidaan huomata näytteissä ongelmalliseksi raskasmetalliksi identifioidun sinkin sitoutuvan parhaiten sementin ja kipsin sekoituksella. Myös tuhkaa sisältävät sideainesekoitukset pienensivät sinkin liukoisuutta verrattuna pelkkään yleis-sementtiin. Jatkotutkimuksissa voitaisiin testata erilaisia sideainesekoituksia betonira-kentamisessa, joilla saadaan ainakin 25 MPa lujuusarvo, pilaantunutta sedimenttiä tai maa-ainesta käyttäen.

Suivi à long terme de l’efficacité d’un traitement de physiothérapie sur l'incontinence urinaire d'effort persistante en période postnatale

L’incontinence urinaire d’effort (IUE) est une condition fréquente en période postnatale pouvant affecter jusqu’à 77% des femmes. Neuf femmes sur dix souffrant d’IUE trois mois après l’accouchement, vont présenter une IUE cinq ans plus tard. Le traitement en physiothérapie de l’IUE par le biais d’un programme d’exercices de renforcement des muscles du plancher pelvien est reconnu comme étant un traitement de première ligne efficace. Les études ont prouvé l’efficacité de cette approche sur l’IUE persistante à court terme, mais les résultats de deux ECR à long terme n’ont pas démontré un maintien de l’effet de traitement. L’effet d’un programme en physiothérapie de renforcement du plancher pelvien intensif et étroitement supervisé sur l’IUE postnatale persistante avait été évalué lors d’un essai clinique randomisé il y a sept ans. Le but principal de la présente étude était d’évaluer l’effet de ce programme sept ans après la fin des interventions de l’ECR initial. Un objectif secondaire était de comparer l’effet de traitement à long terme entre un groupe ayant fait seulement des exercices de renforcement du plancher pelvien et un groupe ayant fait des exercices de renforcement du plancher pelvien et des abdominaux profonds. Un troisième objectif était d’explorer l’influence de quatre facteurs de risques sur les symptômes d’IUE et la qualité de vie à long terme. Les cinquante-sept femmes ayant complétées l’ECR initial ont été invitées à participer à l’évaluation du suivi sept ans. Vingt et une femmes ont participé à l’évaluation clinique et ont répondu à quatre questionnaires, tandis que dix femmes ont répondu aux questionnaires seulement. L’évaluation clinique incluait un pad test et la dynamométrie du plancher pelvien. La mesure d’effet primaire était un pad test modifié de 20 minutes. Les mesures d’effets secondaires étaient la dynamométrie du plancher pelvien, les symptômes d’IUE mesuré par le questionnaire Urogenital Distress Inventory, la qualité de vie mesurée par le questionnaire Incontinence Impact Questionnaire et la perception de la sévérité de l’IUE mesuré par l’Échelle Visuelle Analogue. De plus, un questionnaire portant sur quatre facteurs de risques soit, la présence de grossesses subséquentes, la v présence de constipation chronique, l’indice de masse corporel et la fréquence des exercices de renforcement du plancher pelvien de l’IUE, venait compléter l’évaluation. Quarante-huit pour-cent (10/21) des participantes étaient continentes selon de pad test. La moyenne d’amélioration entre le résultat pré-traitement et le suivi sept ans était de 26,9 g. (écart-type = 68,0 g.). Il n’y avait pas de différence significative des paramètres musculaires du plancher pelvien entre le pré-traitement, le post-traitement et le suivi sept ans. Les scores du IIQ et du VAS étaient significativement plus bas à sept ans qu’en prétraitement (IIQ : 23,4 vs 15,6, p = 0,007) et (VAS : 6,7 vs 5,1, p = 0,001). Les scores du UDI étaient plus élevés au suivi sept ans (15,6) qu’en pré-traitement (11,3, p = 0,041) et en post-traitement (5,7, p = 0,00). La poursuite des exercices de renforcement du plancher pelvien à domicile était associée à une diminution de 5,7 g. (p = 0,051) des fuites d’urine observées au pad test selon une analyse de régression linéaire. Les limites de cette étude sont ; la taille réduite de l’échantillon et un biais relié au désir de traitement pour les femmes toujours incontinentes. Cependant, les résultats semblent démontrer que l’effet du traitement à long terme d’un programme de renforcement des muscles du plancher pelvien qui est intensif et étroitement supervisé, est maintenu chez environ une femme sur deux. Bien que les symptômes d’IUE tel que mesuré par les pad test et le questionnaire UDI, semblent réapparaître avec le temps, la qualité de vie, telle que mesurée par des questionnaires, est toujours meilleure après sept qu’à l’évaluation initiale. Puisque la poursuite des exercices de renforcement du plancher pelvien est associée à une diminution de la quantité de fuite d’urine au pad test, les participantes devraient être encouragées à poursuivre leurs exercices après la fin d’un programme supervisé. Pour des raisons de logistique la collecte de donnée de ce projet de recherche s’est continuée après la rédaction de ce mémoire. Les résultats finaux sont disponibles auprès de Chantale Dumoulin pht, PhD., professeure agrée à l’Université de Montréal.

Prevalence of Leptospira species among farmed and domestic animals in Greece

A total of 1527 serum samples from pigs, goats, sheep, cattle and dogs in Greece were examined by the microscopic agglutination test and 11.8 per cent of them had antibodies against one or more Leptospira serovars at titres of 1/100 or more. The predominant serovar affecting farm animal species was Bratislava, and Copenhageni was common among dogs and the second most important serovar when all animals were considered together. Another prevalent serovar was Australis, but antibodies to Pomona were detected only in goats and cattle.

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar-gel immunodiffusion (ACID) test for canine brucellosis, were used as the control panel for B. cants infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies. (C) 2010 Elsevier Ltd. All rights reserved.

Ovine leptospirosis in Brazil

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

Serosurvey for antibodies against Brucella abortus and Leptospira interrogans in pampas deer from Brazil

A survey for antibodies against Brucella abortus, and Leptospira interrogans was conducted on 17 pampas deer (Ozotocerus bezoarticus) from Pantanal Matogrossense (State of Mato Grosso do Sul, Brazil) and on 24 pampas deer from Parque Nacional de Emas (State of Goias, Brazil). Antibodies against B. abortus were detected by plate ag glutination, rose Bengal, and complement fixation tests, antibodies against Leptospira interrogans were detected by the microscopic agglutination test. All sera were negative for B. abortus antibodies and all deer sera from Parque Nacional de Emas were negative for L. interrogans antibodies. Four (24%) of 17 sera from Pantanal Matogrossense were positive for L. interrogans serovar (n = 2) hardjo, wolffi (n = 1) and mini (n = 1). While these diseases do not appear to be of major importance to the health status of Pampas deer, it appears that deer are reservoir for leptospirosis in one of the study areas.

Use of saprophytic leptospira strains in the serodiagnosis of experimental leptospirosis in guinea-pigs (Cavia sp)

No presente estudo foi avaliada, através da reação de soroaglutinação microscópica, a eficiência de quatro estirpes apatogênicas de Leptospira biflexa (Buenos Aires, Patoc 1, Rufino e São Paulo) como antígeno único para o diagnóstico sorológico em cobaias experimentalmente infectadas com sete diferentes sorotipos de Leptospira interrogans (canicola, grippotyphosa, hardjo, icterohaemorrhagiae, pomona, tarassovi e wolffi). As estirpes Buenos Aires, Patoc 1, Rufino e São Paulo não foram eficientes em revelar títulos de anticorpos nos soros das cobaias infectadas com Leptospira interrogans. Durante o experimento, foi observada a ocorrên cia de reações sorológicas cruzadas entre as estirpes Patoc 1 e São Paulo e também entre os sorotipos wolffi e hardjo.