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The present study aimed to investigate the occurrence of Mycoplasma synoviae (MS), M. gallisepticum (MG), Ornitobacterium rhinotracheale (OR), Avibacterium paragallinarum (AP), Pasteurella multocida (PM) and Infectious Bronchitis Virus (IBV) in laying hens with respiratory clinical signs in two phases of production. 140 tracheal swabs and 140 blood samples were collected from laying hens in the rearing and production phases, the chickens belonged to six farms (A-F) located in the state of São Paulo, Brazil. The samples were analyzed by PCR for MG, MS, OR, AP, PM and IBV and by ELISA for MG and MS. The highest frequencies observed by PCR were for MS at farms B and C with 95 and 100% positivity, followed by MG at farms D and E with 35% and 65%, IBV with 35% at farm F and ORT with 15% at farm A. All flocks were positive for MG and MS in serology. Although MG and IBV have been detected, this can be explained by the vaccination protocols, since live attenuated vaccines are widely used for immunization against these pathogens. It was also possible to detect OR and AP thorugh PCR in some flocks. The occurrence of several etiological agents that cause respiratory diseases in laying hens was confirmed by PCR and serology, with MS being the most prevalent and being present in all farms studied.(AU)

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RESUMO

The present study aimed to investigate the occurrence of Mycoplasma synoviae (MS), M. gallisepticum (MG), Ornitobacterium rhinotracheale (OR), Avibacterium paragallinarum (AP), Pasteurella multocida (PM) and Infectious Bronchitis Virus (IBV) in laying hens with respiratory clinical signs in two phases of production. 140 tracheal swabs and 140 blood samples were collected from laying hens in the rearing and production phases, the chickens belonged to six farms (A-F) located in the state of São Paulo, Brazil. The samples were analyzed by PCR for MG, MS, OR, AP, PM and IBV and by ELISA for MG and MS. The highest frequencies observed by PCR were for MS at farms B and C with 95 and 100% positivity, followed by MG at farms D and E with 35% and 65%, IBV with 35% at farm F and ORT with 15% at farm A. All flocks were positive for MG and MS in serology. Although MG and IBV have been detected, this can be explained by the vaccination protocols, since live attenuated vaccines are widely used for immunization against these pathogens. It was also possible to detect OR and AP thorugh PCR in some flocks. The occurrence of several etiological agents that cause respiratory diseases in laying hens was confirmed by PCR and serology, with MS being the most prevalent and being present in all farms studied.

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O objetivo deste trabalho foi estudar a prevalência de MG e MS e a filogenia das cepas circulantes, comparando-as com outras já descritas em poedeiras comerciais no Brasil. Foram coletados 140 suabes traqueais de poedeiras comerciais com sinais respiratórios em seis granjas da região centro-oeste de São Paulo. As amostras foram avaliadas por PCR, com posterior sequenciamento e análise filogenética das cepas identificadas. Das 140 amostras, 16,4% foram positivas para MG e 68,6% para MS. Houve diferença significativa nas frequências de MG e MS por granja, segundo o teste G de independência (P<0,05). Todas as cepas identificadas de MG e MS de granjas distintas apresentaram similaridade tanto pela lipoproteína para MG quanto pela região 16s rRNA para MS. Neste estudo, foi possível observar altas prevalências dos agentes estudados, sendo a de MS maior que a de MG. Foi detectada infecção mista por MG e MS em 11,4% das amostras e sabe-se que esses micoplasmas podem agir de forma sinérgica, agravando o quadro respiratório. As cepas circulantes identificadas, pela análise das regiões gênicas da lipoproteína para MG e 16S rRNA para MS, são similares em todas as granjas estudadas.(AU)

The aim of this study was to evaluate the prevalence of MG and MS and the phylogeny of the circulating strains, comparing them with others already described in commercial laying hens from Brazil. A total of 140 tracheal swabs were collected from commercial laying hens with respiratory signs in six farms from the western region of São Paulo state. The samples were analyzed by PCR with subsequent sequencing and phylogenetic analysis of the identified strains. From the 140 samples, 68.6% were positive for MS and 16.4% for MG. There was a significant difference in the frequencies of MG and MS per farm according to G Test of independence (P<0.05). All strains identified as MG and MS from distinct farms presented similarity both by lipoprotein to MG and by 16s rRNA region to MS. In this study, it was possible to observe a high prevalence of MS compared to MG. Mixed MG and MS infection was detected in 11.4% of the samples. These mycoplasmas may act synergistically, worsening the respiratory signs. The circulating strains identified by analysis of the lipoprotein for MG and 16S rRNA for MS are similar on all poultry farms studied.(AU)

Assuntos

RESUMO

O objetivo deste trabalho foi estudar a prevalência de MG e MS e a filogenia das cepas circulantes, comparando-as com outras já descritas em poedeiras comerciais no Brasil. Foram coletados 140 suabes traqueais de poedeiras comerciais com sinais respiratórios em seis granjas da região centro-oeste de São Paulo. As amostras foram avaliadas por PCR, com posterior sequenciamento e análise filogenética das cepas identificadas. Das 140 amostras, 16,4% foram positivas para MG e 68,6% para MS. Houve diferença significativa nas frequências de MG e MS por granja, segundo o teste G de independência (P<0,05). Todas as cepas identificadas de MG e MS de granjas distintas apresentaram similaridade tanto pela lipoproteína para MG quanto pela região 16s rRNA para MS. Neste estudo, foi possível observar altas prevalências dos agentes estudados, sendo a de MS maior que a de MG. Foi detectada infecção mista por MG e MS em 11,4% das amostras e sabe-se que esses micoplasmas podem agir de forma sinérgica, agravando o quadro respiratório. As cepas circulantes identificadas, pela análise das regiões gênicas da lipoproteína para MG e 16S rRNA para MS, são similares em todas as granjas estudadas.(AU)

The aim of this study was to evaluate the prevalence of MG and MS and the phylogeny of the circulating strains, comparing them with others already described in commercial laying hens from Brazil. A total of 140 tracheal swabs were collected from commercial laying hens with respiratory signs in six farms from the western region of São Paulo state. The samples were analyzed by PCR with subsequent sequencing and phylogenetic analysis of the identified strains. From the 140 samples, 68.6% were positive for MS and 16.4% for MG. There was a significant difference in the frequencies of MG and MS per farm according to G Test of independence (P<0.05). All strains identified as MG and MS from distinct farms presented similarity both by lipoprotein to MG and by 16s rRNA region to MS. In this study, it was possible to observe a high prevalence of MS compared to MG. Mixed MG and MS infection was detected in 11.4% of the samples. These mycoplasmas may act synergistically, worsening the respiratory signs. The circulating strains identified by analysis of the lipoprotein for MG and 16S rRNA for MS are similar on all poultry farms studied.(AU)

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Fowl paratyphoid infections are caused by different Salmonella serovars that can affect a wide range of hosts. Due to its complex epidemiology, Salmonella serovar identification is crucial for the development and implementation of monitoring and control programs in poultry farms. Moreover, the characterization of the antimicrobial resistance profiles of Salmonella strains isolated from livestock is relevant to public health because they are a common causative agent of foodborne diseases. The objective of this study was to investigate the presence of Salmonella spp. and to identify the antimicrobial resistance profiles of strains isolated in the midwestern region of São Paulo state, which accounts for the highest production of table eggs in Brazil. For this purpose, 2008 fecal samples were collected on 151 commercial layer farms and submitted to microbiological analyses. Twenty-two serovars were isolated from 80 (52.9%) farms, among which S. Mbandaka and S. Braenderup were the most prevalent. All isolates expressed resistance to at least one of the 23 antimicrobials tested, and the highest resistance rates were determined against streptomycin (93.5%) and sulfonamide (84.6%). Moreover, multidrug resistance was observed in 41% of the isolates and the maximum drug resistance profile was against ten different antimicrobials. Therefore, the identification of Salmonella serovars in poultry production provides epidemiological knowledge to develop prevention and control measures in order to ensure poultry health and to prevent human infection by multiresistant strains.(AU)

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Fowl paratyphoid infections are caused by different Salmonella serovars that can affect a wide range of hosts. Due to its complex epidemiology, Salmonella serovar identification is crucial for the development and implementation of monitoring and control programs in poultry farms. Moreover, the characterization of the antimicrobial resistance profiles of Salmonella strains isolated from livestock is relevant to public health because they are a common causative agent of foodborne diseases. The objective of this study was to investigate the presence of Salmonella spp. and to identify the antimicrobial resistance profiles of strains isolated in the midwestern region of São Paulo state, which accounts for the highest production of table eggs in Brazil. For this purpose, 2008 fecal samples were collected on 151 commercial layer farms and submitted to microbiological analyses. Twenty-two serovars were isolated from 80 (52.9%) farms, among which S. Mbandaka and S. Braenderup were the most prevalent. All isolates expressed resistance to at least one of the 23 antimicrobials tested, and the highest resistance rates were determined against streptomycin (93.5%) and sulfonamide (84.6%). Moreover, multidrug resistance was observed in 41% of the isolates and the maximum drug resistance profile was against ten different antimicrobials. Therefore, the identification of Salmonella serovars in poultry production provides epidemiological knowledge to develop prevention and control measures in order to ensure poultry health and to prevent human infection by multiresistant strains.

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Alcoholic patients are more susceptible to Strongyloides stercoralis infection. The chronic use of alcohol raises the levels of endogenous corticosteroids, which regulates the development of larvae and stimulates the differentiation of rhabditiform into infective filariform larvae, thus inducing internal autoinfection. Therefore, early diagnosis is important to prevent severe strongyloidiasis. The aim of this study was to evaluate the efficacy of parasitological methods, according to the parasite load and the number of stool samples, for diagnosis of S. stercoralis infection, as well the peripheral blood eosinophil count in alcoholic patients. A total of 330 patients were included in this study. The diagnosis was established using three parasitological methods: agar plate culture, Baermann-Moraes method and spontaneous sedimentation. Peripheral eosinophilia was considered when the level was >600 eosinophils/mm3. The agar plate culture (APC) had the highest sensitivity (97.3%). However, the analysis of multiple samples increased the sensitivity of all parasitological methods. The sensitivities of the methods were influenced by the parasite load. When the larval number was above 10, the sensitivity of APC was 100%, while in spontaneous sedimentation the sensitivity reached 100% when the larval number was above 50. In the present study, 15.4% of alcoholic patients infected with S. stercoralis (12/78) had increased peripheral blood eosinophil count (above 600 eosinophils/mm3). For an efficient parasitological diagnosis of S. stercoralis infection in alcoholic patients, repeated examination by two parasitological methods must be recommended, including agar plate culture due to its higher sensitivity. Moreover, S. stercoralis infection was associated with eosinophilia, mostly in patients excreting up to 10 larvae/g faeces.

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The mite Allopsoroptoides galli has recently been identified parasitizing commercial chickens, São Paulo State/Brasil, causing severe dermatitis on all parts of the animal's body and a significant decline in productivity, particularly in egg production. The aim of the present study in A. galli infestation was to investigate the impact on laying hens' performance and egg quality. A total of 100 56-week-old Hy-line white laying hens were used. The birds were divided into 2 groups, with 10 replicates of 5 birds in each group. The experimental groups consisted of a non-infested group (hens free of theA. galli) and an infested group (hens presenting A. galli). The infestation with A. galli did not significantly influence feed intake but caused a significant reduction in the body weight of the hens and caused a decrease in egg production, therefore promoting worse feed conversion. The egg weight was reduced; however, the infestation did not significantly affect the internal quality of the eggs, which was measured according to the yolk color, albumen height, and Haugh units, or the quality of the shell, based on its percentage, thickness, and strength. It can be concluded that anA. galli infestation promotes a reduction in body weight, egg production, and egg weight in laying hens, therefore worsening feed conversion.

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Mycoplasma synoviae infection of hens has been associated with problems of eggshell quality called eggshell apex abnormalities (EAA). Little is known about the quality of EAA eggs from a commercial point of view, especially during their storage. The study aimed to examine the differences between EAA and normal eggs during storage under controlled conditions in 2 seasons, summer and winter, by comparing internal and external quality parameters. In a conventional egg production farm with white laying hens of varying ages in the city of Bastos, state of São Paulo, Brazil, 232 eggs were used in the summer season and 400 eggs in the winter season. Half of the eggs had EAA, and the other half were considered normal eggs for each season. The eggs were analyzed at 2, 7, 14, 21, and 28 d after being laid and stored from 24.6 to 25.8°C in summer and from 24 to 25°C in winter. There was no difference (P > 0.05) in the average egg weight between EAA and normal eggs at any studied time point, but in both seasons, the weight loss in EAA eggs was higher than in normal eggs. The losses in Haugh unit scores from the first to the last measurements were approximately 40% regardless of egg type or season of production. In comparing eggshell thickness, only the apices of the EAA eggs were thinner (P < 0.0001) than normal eggs in the summer, but in the winter, the EAA egg apices (P < 0.0001) and sides (P = 0.03) were both thinner. The presence of EAA did not affect the eggshell weight (P > 0.05) or eggshell percentage (P > 0.05). The eggshell strength of the EAA eggs was lower (P < 0.0001) than normal eggs in both the summer (16.57%) and winter (19.86%). The presence of EAA did not affect the internal quality of the egg, but was related to a greater loss of external quality and weight during storage.

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The aim of this study was to evaluate whether the drinking water of the School of Agricultural and Veterinary Sciences, UNESP, Jaboticabal, Brazil, affected bone mineral density and serum calcium levels of 14-, 21-, and 45-day-old broilers. Bone mineral density (BMD) of the tibiae was assessed using optical densitometry radiographic technique and serum calcium levels. Tibial BMD increased as broilers aged, and achieved its peak at 45 days of age. It was higher in the distal epiphysis of the birds that ingested filtered water (p 0.05) compared with those supplied with unfiltered water. Therefore, it is concluded that filtered water promoted better bone quality in relative to those ingested unfiltered water.(AU)

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The aim of this study was to evaluate whether the drinking water of the School of Agricultural and Veterinary Sciences, UNESP, Jaboticabal, Brazil, affected bone mineral density and serum calcium levels of 14-, 21-, and 45-day-old broilers. Bone mineral density (BMD) of the tibiae was assessed using optical densitometry radiographic technique and serum calcium levels. Tibial BMD increased as broilers aged, and achieved its peak at 45 days of age. It was higher in the distal epiphysis of the birds that ingested filtered water (p 0.05) compared with those supplied with unfiltered water. Therefore, it is concluded that filtered water promoted better bone quality in relative to those ingested unfiltered water.

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In the present study Salmonella spp. was surveyed in four flocks of meat-type quails reared in a farm that also had processing plant on site, located in the region of Bastos, state of São Paulo, Brazil. Meconium samples of one-day-old quail chicks were collected from transport cardboard boxes. Cecal content was collected on days 7, 14, 21, 28 and 35 of rearing. At 36 days of age, birds were slaughtered in the farm's processing plant, where two samples of water from the scalding and the chilling tanks and four carcasses per flock were collected. All samples were examined for Salmonella spp. using traditional bacteriological methods. Salmonella spp. was present in meconium samples of three flocks and in cecal feces of the four flocks. This bacterium was also isolated in the chiller water and in the carcasses of three of the evaluated flocks and in the scalding water of one flock. In this study, S. enterica subspecies enterica 4, 5, 12; S. Corvalis; S. Give; S. Lexington; S. Minnesota; S. Schwarzengrund; S. Rissen and S. Typhimurium were the eight serovars identified.(AU)

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In the present study Salmonella spp. was surveyed in four flocks of meat-type quails reared in a farm that also had processing plant on site, located in the region of Bastos, state of São Paulo, Brazil. Meconium samples of one-day-old quail chicks were collected from transport cardboard boxes. Cecal content was collected on days 7, 14, 21, 28 and 35 of rearing. At 36 days of age, birds were slaughtered in the farm's processing plant, where two samples of water from the scalding and the chilling tanks and four carcasses per flock were collected. All samples were examined for Salmonella spp. using traditional bacteriological methods. Salmonella spp. was present in meconium samples of three flocks and in cecal feces of the four flocks. This bacterium was also isolated in the chiller water and in the carcasses of three of the evaluated flocks and in the scalding water of one flock. In this study, S. enterica subspecies enterica 4, 5, 12; S. Corvalis; S. Give; S. Lexington; S. Minnesota; S. Schwarzengrund; S. Rissen and S. Typhimurium were the eight serovars identified.

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This work aimed at investigating the lipid profile of zoonotic visceral leishmaniasis (VL) patients' sera and the effect of lipoproteins on the in vitro production of tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-6, IL-10 and IL-12 by Leishmania infantum-infected and uninfected macrophages. Lipids were quantified in 26 VL patients' sera and 26 healthy controls from a VL endemic area. The patients' sera had higher triglyceride and very low density lipoprotein (VLDL) levels, and much lower apolipoprotein A1, total cholesterol, low density lipoprotein (LDL) and high density lipoprotein (HDL) levels than the control sera. Lipoprotein fractions were obtained by ultracentrifugation of sera. The addition of LDL and HDL to Leishmania-infected and uninfected macrophages, in physiological concentrations, enhanced the production of IL-6 and IL-10, but not of IL-12. LDL stimulated the production of TNF-alpha only in infected macrophages, whereas HDL stimulated the production of lower amounts of TNF-alpha in both infected and uninfected macrophages. VLDL stimulated only the production of IL-10. It is proposed herein that LDL may influence the development of VL by promoting the production of TNF-alpha by infected macrophages. A decrease in plasma LDL in some VL patients (to 20 mg/mL or less); however, would tend to reduce the production of TNF-alpha and therefore to limit the development of immune-mediated pathology, not withstanding the fact that it would perhaps increase the permissiveness of macrophages to Leishmania growth.

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Opportunistic infections, due to disease-related immunosuppression, constitute the major cause of death in American visceral leishmaniasis (AVL). Sera from these patients (AVL sera) non-specifically inhibit the in vitro proliferative response of normal human lymphocytes to lectins or antigens. In the present work, the mediation of this inhibition by IgG, immune complexes and low- or very low-density lipoproteins was studied. AVL serum fractions containing proteins with the molecular weight of IgG, and IgG, purified from AVL sera by anion exchange chromatography, did not suppress the lymphoproliferation. Most of the suppressive activity of AVL sera was associated with a fraction containing molecules with molecular weights above 430 kDa. This would be compatible with it being due to immune complexes and/or lipoproteins, and not to soluble IL-2 receptors as reported previously. However, neither of the two possibilities seem to be the case, as (1) depletion of immune complexes by protein-A followed by protein-G chromatographies did not affect the serum suppressive activity, (2) no correlation between immune complex contents and suppressive activities in individual sera was observed, and (3) plasma lipoproteins (VLDL and LDL), purified from AVL patients and from healthy individuals, had the same degree of immunosuppressive activity.

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The present work compares six biochemical methods for extraction of lipids from human serum. Although some organic solvents were good lipid extractors, they precipitated most of the total proteins and albumin. On the other hand, methodologies using Triton X-114 and silica were efficient for extraction of lipids, while sparing the protein fraction.

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A semi-quantitative ELISA for complement-fixing, IgG-containing immune complexes (IC) is described. The assay is based on the insolubilization of IC by polyethyleneglycol, their capture by solid-phase anti-C3 antibodies, reaction with peroxidase-labeled anti-IgG antibodies and incubation with a chromogenic peroxidase substrate. It was markedly improved by the use of a single-step procedure which simultaneously washed and precipitated the insolubilized immune complexes. Intra-assay and inter-assay coefficients of variation were lower than 8.6 and 14.7%, respectively. As expected, higher levels of circulating immune complexes, in relation to healthy individuals, were found in patients with American visceral leishmaniasis (AVL), systemic lupus erythematosus (SLE) and rheumatoid arthritis (RA), with prevalences comparable to those described in the literature. The ELISA can be quickly assembled from reagents and plasticware widely available commercially, detects immune complexes fulfilling three different criteria and is more sensitive than a previously published method based on the same principles (detection limit for complement-sensitized aggregated IgG of 2 microg ml(-1) as compared with a detection limit above 16 microg ml(-1)).

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