Evaluation of a MPB70-ELISA to differentiate Mycobacterium bovis from M. avium-sensitized swine

Swine are susceptible to different mycobacteria species, being Mycobacterium bovis an agent of tuberculosis, with most significant zoonotic risks, while M. avium determines a granulomatous lymphadenitis with low zoonotic risk. Currently performed intradermal tests present some important limitations, such as the lack of ability to detect anergic animals or to differentiate among mycobacterial species. In order to improve the TB diagnosis, serological assays have been developed, with encouraging results. The purpose of this study was to evaluate the performance of a MPB70-ELISA in 82 piglets divided into four groups: sensitized by inactivated M. bovis, M. avium, inoculated with oil adjuvant, or with saline solution. The test was able to discriminate between an animal sensitized by M. bovis and animals of the three other groups, including M. avium-sensitized animals; for this reason, we suggest that MPB70-ELISA could be used as a complementary tool for discriminating the agent of the mycobacteriosis, and therefore to diagnose tuberculosis in a swine herd.

First isolation and characterization of Leptospira interrogans serogroup Australis from swine in Brazil

The purpose of this study was to report the first recovery and characterization of Leptospira interrogans (serogroup Australis) from urine of swine in Brazil. The isolate was studied by serogrouping, MLVA, PGFE, and partial sequencing of rrs and secY. It was serogrouped as serogroup Australis, probably serovar Bratislava (titre 1,600), and sequenced as Leptospira interrogans. The MLVA and PGFE profiles also suggested the isolate as serovar Bratislava, since they were indistinguishable from reference strains Balico and Jez Bratislava. This is the first Leptospira interrogans serogroup Australis isolate, probably serovar Bratislava, obtained in Brazil.

The role of alveolar type II cells in swine leptospirosis

Abstract: This study aimed to investigate a possible relationship between alveolar type II cells and the inflammatory response to infection with Leptospira spp., and thus comprise a further element that can be involved in the pathogenesis of lung injury in naturally infected pigs. The study group consisted of 73 adult pigs that were extensively reared and slaughtered in Teresina, Piauí state, and Timon, Maranhão state, Brazil. The diagnosis of leptospirosis was made using the microscopic agglutination test (MAT) aided by immunohistochemistry and polymerase chain reaction. The MAT registered the occurrence of anti-Leptospira antibodies in 10.96% (8/73) of the pigs. Immunohistochemistry allowed for the visualization of the Leptospira spp. antigen in the lungs of 87.67% (64/73) of the pigs. There was hyperplasia of bronchus-associated lymphoid tissue and circulatory changes, such as congestion of alveolar septa, parenchymal hemorrhage and edema within the alveoli. Lung inflammation was more intense (p = 0.0312) in infected animals, which also showed increased thickening of the alveolar septa (p = 0.0006). Evaluation of alveolar type II (ATII) cells using an anti-TTF-1 (Thyroid Transcription Factor-1) antibody showed that there were more immunostained cells in the non-infected pigs (53.8%) than in the infected animals (46.2%) and that there was an inverse correlation between TTF-1 positive cells and the inflammatory infiltrate. There was no amplification of Leptospira DNA in the lung samples, but leptospiral DNA amplification was observed in the kidneys. The results of this study showed that a relationship exists between a decrease in alveolar type II cells and a leptospire infection. Thus, this work points to the importance of studying the ATII cells as a potential marker of the level of lung innate immune response during leptospirosis in pigs.

Gastrointestinal parasites of swine raised in different management systems in the State of Rio de Janeiro, Brazil

Abstract: From 2012 to 2013 were surveyed gastrointestinal parasites from pig farms located in different municpaliyies in the state of Rio de Janeiro. Fecal samples from 790 pigs were collected from the rectum on 88 family farms and 702 farms with industrial production. The samples were subjected to Faust et al., Sheather, Ritchie, Lutz and direct examination faecal techniques. The estimated parasite prevalence was 93.1% in family farms and 59.1% in industrial farms. Balantidium coli, coccidia and Entamoeba sp. were the parasites with the highest frequencies, and the male and female reproductive categories and fatteners pigs the most infected (p<0.05). Trophozoites of B. coli were most evident in stool samples from semi-solid followed by solid and diarrheal consistencies. Strongyles eggs and Trichuris suis have been detected exclusively in family farms. Ascaris suum eggs and Strongyloides ransomi showed low frequency. The high degree of parasitism, especially protozoa, indicates the need to reassess the management of pigs in both types of production.

Effect of psychological stress on the L-arginine-nitric oxide pathway and semen quality

It has been reported that mental stress causes abnormality of spermiogram parameters. We investigated the effect of psychological stress on the L-arginine-nitric oxide (NO) pathway. Semen samples were collected from 29 healthy fourth semester medical students just before (stress) and 3 months after (non-stress) the final examinations. Psychological stress was measured by the State Anxiety Inventory questionnaire. After standard semen analysis, arginase activity and NO concentration were measured spectrophotometrically in the seminal plasma. Measurements were made in duplicate. During the stress period, sperm concentration (41.28 ± 3.70 vs 77.62 ± 7.13 x 10(6)/mL), rapid progressive motility of spermatozoa (8.79 ± 1.66 vs 20.86 ± 1.63%) and seminal plasma arginase activity (0.12 ± 0.01 vs 0.22 ± 0.01 U/mL) were significantly lower than in the non-stress situation, whereas seminal plasma NO (17.28 ± 0.56 vs 10.02 ± 0.49 µmol/L) was higher compared to the non-stress period (P < 0.001 for all). During stress there was a negative correlation between NO concentration and sperm concentration, the percentage of rapid progressive motility and arginase activity (r = -0.622, P < 0.01; r = -0.425, P < 0.05 and r = -0.445, P < 0.05, respectively). These results indicate that psychological stress causes an increase of NO level and a decrease of arginase activity in the L-arginine-NO pathway. Furthermore, poor sperm quality may be due to excessive production of NO under psychological stress. In the light of these results, we suggest that the arginine-NO pathway, together with arginase and NO synthase, are involved in semen quality under stress conditions.

Development and validation of a genotype 3 recombinant protein-based immunoassay for hepatitis E virus serology in swine

Hepatitis E virus (HEV) is classified within the family Hepeviridae, genus Hepevirus. HEV genotype 3 (Gt3) infections are endemic in pigs in Western Europe and in North and South America and cause zoonotic infections in humans. Several serological assays to detect HEV antibodies in pigs have been developed, at first mainly based on HEV genotype 1 (Gt1) antigens. To develop a sensitive HEV Gt3 ELISA, a recombinant baculovirus expression product of HEV Gt3 open reading frame-2 was produced and coated onto polystyrene ELISA plates. After incubation of porcine sera, bound HEV antibodies were detected with anti-porcine anti-IgG and anti-IgM conjugates. For primary estimation of sensitivity and specificity of the assay, sets of sera were used from pigs experimentally infected with HEV Gt3. For further validation of the assay and to set the cutoff value, a batch of 1100 pig sera was used. All pig sera were tested using the developed HEV Gt3 assay and two other serologic assays based on HEV Gt1 antigens. Since there is no gold standard available for HEV antibody testing, further validation and a definite setting of the cutoff of the developed HEV Gt3 assay were performed using a statistical approach based on Bayes' theorem. The developed and validated HEV antibody assay showed effective detection of HEV-specific antibodies. This assay can contribute to an improved detection of HEV antibodies and enable more reliable estimates of the prevalence of HEV Gt3 in swine in different regions.

Cultivation of microalgae Spirulina platensis (Arthrospira platensis) from biological treatment of swine wastewater

The microalgae biomass production from swine wastewater is a possible solution for the environmental impact generated by wastewater discharge into water sources. The biomass can be added to fish feed, which can be used in the formulation of meat products. This work addresses the adaptation of the microalgae Spirulina platensis (Arthrospira platensis) in swine wastewater and the study of the best dilution of the wastewater for maximum biomass production and for removal of Chemical Oxygen Demand (COD), ammonia and phosphorous to the microalgae. The cultivation of Spirulina platensis, strain Paracas presented maximum cellular concentrations and maximum specific growth rates in the wastewater concentration of 5.0 and 8.5%. The highest COD removals occurred with 26.5 and 30.0% of wastewater in the medium. The maximum removal of total phosphorous (41.6%), was with 8.5% of wastewater, which is related to the microalgae growth. The results of Spirulina culture in the swine wastewater demonstrated the possibility of using these microalgae for the COD and phosphorous removal and for biomass production.

Opresión internalizada y prácticas sexuales de riesgo en varones homo-y bi-sexuales de México

OBJETIVO: Analizar la relación entre prácticas sexuales de riesgo para infección por VIH y la opresión que éstos han internalizado debido al prejuicio y discriminación que se enfrentan varones bisexuales y homosexuales. MÉTODOS: Estudio observacional, transversal, y analítico. Entre septiembre y noviembre del 2001 se aplicó un cuestionario a 318 hombres que acudieron a organizaciones e instituciones ubicadas en la Ciudad de México. Las variables indagadas fueron: homofobia internalizada, percepción del estigma, ocultamiento y prácticas sexuales en los seis meses previos a la encuesta. Mediante modelos de regresión logistica se estimó la relación (odds ratios) de las formas de opresión internalizada con las prácticas sexuales de riesgo, ajustando por variables confusoras. RESULTADOS: De los entrevistados, 256 habían tenido por lo menos una relación sexual en los últimos seis meses, 50 respondieron no haberlas tenido y 12 no contestaron esta pregunta. Más del 30% de los varones no había utilizado preservativo cuando habían tenido sexo insertivo o receptivo. La homofobia internalizada se asoció con mayor riesgo de consumo de drogas antes o durante las relaciones sexuales, sexo insertivo sin condón, sexo receptivo sin condón y el tragar semen de otros hombres. CONCLUSIONES: Sigue siendo frecuente la adopción de prácticas sexuales de riesgo entre los varones bi y homosexuales. De las tres formas de opresión internalizada que se investigaron, la que se asoció con mayor probabilidad de prácticas sexuales de riesgo fue la homofobia internalizada. Esta información puede ser incorporada en el diseño de programas de promoción de uso de condón entre varones bi y homosexuales.

Serologic studies on the behaviour of influenza virus type A in persons of greater São Paulo during 1976, 1978 and 1979

Sera of persons of different age groups collected in 1976, 1978 and 1979 were tested for the presence of HI antibodies against various strains of the H3N2 and H1N1 subtypes of influenza virus. The occurrence of infection by H3N2 subtype was recorded during the 1976-1978 period but in 1979, circulation of this subtype of virus was limited. The prevalence of antibody against A/São Paulo/1/78 (H1N1) was significantly higher than that of antibody to A/USSB/90/77 (HIND in 1978. However in 1979 the predominant strain was A/USSR/90/77 (HIND. Persons under 20 years of age were the most affected by H1N1 subtype. Antibodies to H1N1 subtype were detected in sera of individuals older than 20 years in 1976, before the re-emergence of this strain. Serological results indicate that infections by H3N2 subtype in 1978 occurred in 65.4% of young children (0-4 year group). About 47.0% of children from the 0-4 year group had antibodies to H1N1 subtype in 1979. Antibodies to swine influenza virus were detected in 60% of 60+ year old people.

Cysticercus bovis: I. Ensaios de evaginação

Fresh bovine and swine bile and dehidrated bovine bile in association with chloridric acid, glucose, peptone or distilled water were utilized to induce evagination of C. bovis. Better results were obtained by using bovine or swine fresh bile 15% or dehidrated bovine bile 4% in distilled water. The C. bovis evagination pattern was the same in broths containing bovine or swine bile. The bovine dehidrated bile at 4% is better for the evagination than the natural one; in the studied concentrations a increase in the percentage of evagination was observed with a concomitant increase in the concentration of bile.

ELISA test for the diagnosis of cysticercosis in pigs using antigens of Taenia solium and Taenia crassiceps cysticerci

In the present study ELISA was standardized for the diagnosis of swine cysticercosis based on necropsy parameters and confirmed positive and negative control sera. Serum samples from pigs with other infections were also assayed to determine possible cross-reactions. Four antigens were assayed: from Taenia crassiceps vesicular fluid (VF-Tcra) and crude larvae extract (T-Tcra), and from Taenia solium extracts of scolex (S-Ts) and of larvae (T-Ts). A checkerboard evaluation of antigen, serum and conjugate dilutions, as well as the use of Tween-20 and skim cow milk in wash and blocking solution had a marked effect on improving ELISA performance. All the antigens showed a good performance, but VF-Tcra was the best, with 96.0% and 80.0% sensitivities for cut-offs respectively at 2sd and 3sd, and corresponding specificities of 97.5% and 100.0%. Cross-reactivity was observed only with hydatidosis and ascaridiosis. In view of the high performance observed, the ELISA test should be recommended for the diagnosis of cysticercosis in suspected swine in slaughterhouses and for the screening of cysticercosis in swine production. These results will support integrated measures of cysticercosis control throughout the chain of swine production, effectively contributing to public health.

Antigenic typing of brazilian rabies virus samples isolated from animals and humans, 1989-2000

Animal and human rabies samples isolated between 1989 and 2000 were typified by means of a monoclonal antibody panel against the viral nucleoprotein. The panel had been previously established to study the molecular epidemiology of rabies virus in the Americas. Samples were isolated in the Diagnostic Laboratory of the Pasteur Institute and in other rabies diagnostic centers in Brazil. In addition to the fixed virus samples CVS-31/96-IP, preserved in mouse brain, and PV-BHK/97, preserved in cell culture, a total of 330 rabies virus samples were isolated from dogs, cats, cattle, horses, bats, sheep, goat, swine, foxes, marmosets, coati and humans. Six antigenic variants that were compatible with the pre-established monoclonal antibodies panel were defined: numbers 2 (dog), 3 (Desmodus rotundus), 4 (Tadarida brasiliensis), 5 (vampire bat from Venezuela), 6 (Lasiurus cinereus) and Lab (reacted to all used antibodies). Six unknown profiles, not compatible with the panel, were also found. Samples isolated from insectivore bats showed the greatest variability and the most commonly isolated variant was variant-3 (Desmodus rotundus). These findings may be related to the existence of multiple independent transmission cycles, involving different bat species.

The perception, beliefs and practices toward genitourinary schistosomiasis by inhabitants of selected endemic areas (Edo/Delta States) in south-eastern Nigeria

Well-structured questionnaire on the perception, impression and response to genitourinary bilharziasis (Genitourinary schistosomiasis) was administered and explained in local languages: 'Igbo' 'Esan' 'Ezon' Itshekiri and Bini to 33815 inhabitants of selected endemic areas in south-eastern Nigeria from January, 1999 to December, 2001. Out of this number, 3815 (11.3%) were properly filled and returned. About 42.0% of the inhabitants admitted knowledge of the disease, while 14 (0.4%) knew about the aetiologic agent. About 181 (5.0%) who responded, admitted procuring treatment, while 100 (5.0%) declined to seek treatment of any sort. The relationships between water-bodies and human activities, and infection were well discussed. Amongst those who admitted knowledge of the disease but no knowledge of its etiologic agent, declined seeking treatment of any kind, but believe the disease is a natural phenomenon in ones developmental stage and therefore of no morbidity and mortality. Laboratory analysis of urine, faeces, semen and HVS was employed to assess questionnaire responses, and in some cases, physical examination was utilized to augment laboratory analysis in confirming urinal diagnosis. Haematuria was only directly related to egg count in the early part of life. Females were significantly haematuric and excreted more ova than males (p < 0.05). Headache (43.0%) and fever (31.0%) were major clinical signs while sexual pains (22.0%) were the least.

Toxoplasma gondii in fresh pork sausage and seroprevalence in butchers from factories in Londrina, Paraná State, Brazil

The aims of this study were to verify the presence of Toxoplasma gondii cysts in fresh pork sausage and the presence of antibodies against T. gondii in serum of workers from factories with Municipal Inspection Service, in Londrina, PR, Brazil. 149 samples of sausage were collected from eight factories and blood samples from 47 workers. We also took information about the practices that were adopted in the factories and the workers' habits that could influence the prevalence of toxoplasmosis. After bioassay in mice, 13 (8.7%) sausage samples were positive, in one of them T. gondii was isolated and in the other 12 the mice seroconverted. Of 47 workers, 36 (76.6%) worked in sausage production and 11 (23.4%) were involved in other functions; 59.5% (28/47), 55.5% (20/36) and 72.7% (8/11), respectively, had T. gondii antibodies. There were no significant differences in the variables of industries' practices and workers' habits related to T. gondii infection. We concluded that fresh pork sausage could be important in the transmission of toxoplasmosis.

Pig bite in Brazil: a case series from a teaching hospital

A retrospective survey done from 1987 till 1990 revealed that 23 patients bitten by pigs sought medical help at a teaching hospital in Uberlândia, in southeastern Brazil. Most cases (21) were from Uberlândia. The cases were evenly distributed by month and by year; most of them (14/16; 87.5%) occurred between 7. OOa.m. and 7.00 p. m. The male to female ratio was 6.7:1. Age ranged from 6 to 73 (mean 38.95 ± SD 22.06, median 36). The bites were more common on the upper limbs, particularly on the forearms. In 11(47.8%) cases the injury was described as deep. In most cases where information was available the injury was related to capture, transport or immobilisation ofthe pigfor slaughter. The following medical procedures were performed: local cleansing in 19(82.6%) cases, rabies vaccine (12; 52.2%), antirabies serum (2; 8.7%), suturing (6; 26.1%) and tetanus vaccine (12; 52.2%). There was no case of infection at the bite site, neither of rabies or tetanus. By our data, the annual incidence of pig bite in Uberlândia can be estimated to be about 1.5/100.000.