Produção Científica http://repositoriobiologico.com.br//jspui/handle/123456789/1 2026-03-07T00:11:26Z 2026-03-07T00:11:26Z Ninga, Ederina Sapozhnikova, Sapozhnikova Lehotay, Steven J Lightfield, Alan R Monteiro, Sergio H http://repositoriobiologico.com.br//jspui/handle/123456789/1317 2026-02-06T17:14:16Z 2020-05-01T00:00:00Z

Title: High-Throughput Mega-Method for the Analysis of Pesticides, Veterinary Drugs, and Environmental Contaminants by Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry and Robotic Mini-Solid-Phase Extraction Cleanup + Low-Pressure Gas Chromatography-Tandem Mass Spectrometry, Part 2: Catfish Authors: Ninga, Ederina; Sapozhnikova, Sapozhnikova; Lehotay, Steven J; Lightfield, Alan R; Monteiro, Sergio H Abstract: The goal of this study was to develop and validate a new method for simultaneous determination of 106 veterinary drugs and 227 pesticides and their metabolites plus 16 polychlorinated biphenyls (PCBs) at and below their regulatory levels established for catfish muscle in the European Union and U.S.A. To do this, two different QuEChERS-based methods for veterinary drugs and pesticides and PCBs were modified and merged into a single mega-method dubbed “QuEChERSER” (more than QuEChERS), which is presented here for the first time. The mega-method was validated in catfish at four different spiking levels with 10 replicates per level. Sample extraction of 2 g test portions was made with 10 mL of 4:1 (v/v) acetonitrile/water, and then an aliquot was taken for ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) analysis of 106 veterinary drugs and 125 pesticides, including metabolites. The remaining extract after salting out was subjected to automated mini-solid-phase extraction cleanup (Instrument Top Sample Preparation) for immediate injection in low-pressure gas chromatography–tandem mass spectrometry (LPGC–MS/MS). The cleanup was conducted in parallel with the 10 min LPGC–MS/MS analysis for 167 PCBs, pesticides, and metabolites, which was conducted in parallel with the 10 min UHPLC–MS/MS analysis for 231 analytes to increase sample throughput (49 analytes were included in both techniques). In MS/MS, three ion transitions were monitored for nearly all targeted analytes to provide unambiguous identification as well as quantification. Satisfactory recoveries (70–120%) and relative standard deviations of ≤20% were achieved for 98 (92%) of the veterinary drugs and their metabolites and for 222 (91%) of pesticides, metabolites, and PCBs, demonstrating that the developed method is applicable for the analysis of these contaminants in fish as part of regulatory monitoring programs and other purposes. Description: The goal of this study was to develop and validate a new method for simultaneous determination of 106 veterinary drugs and 227 pesticides and their metabolites plus 16 polychlorinated biphenyls (PCBs) at and below their regulatory levels established for catfish muscle in the European Union and U.S.A. To do this, two different QuEChERS-based methods for veterinary drugs and pesticides and PCBs were modified and merged into a single mega-method dubbed “QuEChERSER” (more than QuEChERS), which is presented here for the first time. The mega-method was validated in catfish at four different spiking levels with 10 replicates per level. Sample extraction of 2 g test portions was made with 10 mL of 4:1 (v/v) acetonitrile/water, and then an aliquot was taken for ultra-high-performance liquid chromatography–tandem mass spectrometry (UHPLC–MS/MS) analysis of 106 veterinary drugs and 125 pesticides, including metabolites. The remaining extract after salting out was subjected to automated mini-solid-phase extraction cleanup (Instrument Top Sample Preparation) for immediate injection in low-pressure gas chromatography–tandem mass spectrometry (LPGC–MS/MS). The cleanup was conducted in parallel with the 10 min LPGC–MS/MS analysis for 167 PCBs, pesticides, and metabolites, which was conducted in parallel with the 10 min UHPLC–MS/MS analysis for 231 analytes to increase sample throughput (49 analytes were included in both techniques). In MS/MS, three ion transitions were monitored for nearly all targeted analytes to provide unambiguous identification as well as quantification. Satisfactory recoveries (70–120%) and relative standard deviations of ≤20% were achieved for 98 (92%) of the veterinary drugs and their metabolites and for 222 (91%) of pesticides, metabolites, and PCBs, demonstrating that the developed method is applicable for the analysis of these contaminants in fish as part of regulatory monitoring programs and other purposes.

2020-05-01T00:00:00Z Cabral, Aline Diniz Camargo, Clarice Neves Galleti, Nara Thiers Cacciatori Okuda, Liria Hiromi Pituco, Edviges Maristela Del Fava, Claudia http://repositoriobiologico.com.br//jspui/handle/123456789/1316 2026-02-05T18:12:47Z 2009-01-01T00:00:00Z

Title: Diagnosis of Neospora caninum in bovine fetuses by histology, immunohistochemistry, and nested-PCR Authors: Cabral, Aline Diniz; Camargo, Clarice Neves; Galleti, Nara Thiers Cacciatori; Okuda, Liria Hiromi; Pituco, Edviges Maristela; Del Fava, Claudia Abstract: Neospora caninum, a cause of abortion and stillbirth in cattle, was studied by histology, immunohistochemistry, and nested-PCR, using primers from the Nc5 region of the genomic DNA (PCR PLUS) and primers from the ITS1 region of the ribosomal DNA (PCR JB). A total of 105 fetal samples sent to the Centro de Pesquisa e Desenvolvimento de Sanidade Animal do Instituto Biológico from January 2006 to May 2008 were examined for evidence of N. caninum. Histological examination revealed 71.4% with non-suppurative inflammation in the heart, lung, liver, kidney, placenta, and brain. Immunohistochemistry detected infections in 8.6% of the samples, mainly in the brain, placenta, and heart. Nested-PCR JB revealed 6.7% with infections, while nested-PCR PLUS returned 20.9% positive results, mainly in brain and placenta, and in the pooled liver and heart. Kappa statistics demonstrated little agreement among the three techniques. The three methods are complementary, since they have distinct diagnostic characteristics and were combined to give a positivity rate of 24.8%. Description: Neospora caninum is a protozoan belonging to the phylum Apicomplexa. It was first detected in dogs in Norway (BJERKÅS; MOHN; PRESTHUS, 1984), and was described and named in 1988 by Dubey et al. Studies by McAllister et al. (1998) showed that the domestic dog is a definitive host, as is the coyote (Canis latrans) (GONDIM et al., 2004a). It was subsequently identified as a causal agent of abortion and stillbirth in cattle (THILSTEAD; DUBEY, 1989), and later studies demonstrated its worldwide economic impact (DUBEY; LINDSAY, 1996; ANDERSON et al., 1991).

2009-01-01T00:00:00Z Malaguti, Jane Mary Albinati Cabral, Aline Diniz Abdalla, Raisa Pereira Salgueiro, Yolanda Oliveira Galleti, Nara Thiers Cacciatori Okuda, Liria Hiromi Cunha, Elenice Maria Sequetin Pituco, Edviges Maristela Del Fava, Claudia http://repositoriobiologico.com.br//jspui/handle/123456789/1315 2026-02-05T18:12:32Z 2012-01-01T00:00:00Z

Title: Neospora caninum as causative agent of bovine encephalitis in Brazil Authors: Malaguti, Jane Mary Albinati; Cabral, Aline Diniz; Abdalla, Raisa Pereira; Salgueiro, Yolanda Oliveira; Galleti, Nara Thiers Cacciatori; Okuda, Liria Hiromi; Cunha, Elenice Maria Sequetin; Pituco, Edviges Maristela; Del Fava, Claudia Abstract: For supporting the Brazilian bovine encephalitis surveillance program this study examined the differential diagnosis of Neospora caninum in central nervous system (CNS) by histological analysis (HE staining), immunohistochemistry (IHC), and nested-PCR using a set of primers from the Nc5 region of the genomic DNA and ITS1 region of the ribosomal DNA. A sample of 302 cattle presenting neurological syndrome and negative for rabies, aged 0 to 18 years, from herds in 10 Brazilian states was evaluated for N. caninum from January 2007 to April 2010. All specimens tested negative with IHC and nested-PCR using primers from the ITS1 region of ribosomal DNA, while two positive cases (0.66%) were found using primers from the Nc5 region of genomic DNA: a 20 month-old male and a 72 month-old female, both from São Paulo State. Only the male presented severe multifocal necrotizing encephalitis associated with mononuclear cell infiltration, a pathognomonic lesion caused by parasites of the family Sarcocystidae, and only this case was associated with N. caninum thus representing 0.33% positivity. Future studies should explore the association of IHC and nested-PCR with real-time PCR, a quantitative method that could be standardized for improving the detection of N. caninum in bovine CNS specimens. Description: Neospora caninum is a protozoan belonging to the phylum Apicomplexa, family Sarcocystidae, that mainly causes abortion in cattle, but its impact as causative agent of neurological syndromes has been poorly documented (DUBEY; SCHARES, 2006).

2012-01-01T00:00:00Z Silva, Aline Aparecida da Villalobos, Eliana Monteforte Cassaro Cunha, Elenice Maria Sequetin Lara, Maria do Carmo Custódio de Souza Hunold Nassar, Alessandra Figueiredo de Castro Piatti, Rosa Maria Castro, Vanessa Pinheiro, Eliana Scarcelli Carvalho, Aline Feola de Del Fava, Claudia http://repositoriobiologico.com.br//jspui/handle/123456789/1314 2026-02-05T14:45:41Z 2020-01-01T00:00:00Z

Title: Causes of equine abortion, stillbirth, and perinatal mortality in Brazil Authors: Silva, Aline Aparecida da; Villalobos, Eliana Monteforte Cassaro; Cunha, Elenice Maria Sequetin; Lara, Maria do Carmo Custódio de Souza Hunold; Nassar, Alessandra Figueiredo de Castro; Piatti, Rosa Maria; Castro, Vanessa; Pinheiro, Eliana Scarcelli; Carvalho, Aline Feola de; Del Fava, Claudia Abstract: Abortion and complications in reproduction are important causes of economic loss in horse breeding. Studies of its causal agents can help to identify the primary pathogens or other factors involved and define appropriate measures to reduce its occurrence. This research aimed to investigate the primary causes of equine abortion, stillbirth, and perinatal mortality in regions of Brazil. Tissue from aborted fetuses, stillbirths, neonates and foals submitted to the Biological Institute of São Paulo, Brazil, from January 2010 to July 2013 were processed for viral and bacterial isolation, polymerase chain reaction (PCR), histology, and immunohistochemistry. Bacterial infection was the primary detected cause of abortion, found in 16 of the 53 animals submitted for bacterial analysis followed by viruses analysis in 2 of 105 animals, and noninfectious causes (neonatal isoerythrolysis) in 2 of 105 animals. Fungi were found in a single sample of 53 tested. The most frequent bacteria recovered were Escherichia coli, Enterobacter aerogenes, combined E. coli and Streptococcus spp., Staphylococcus spp., and Bacillus spp. The following agents were each observed in a single sample: Arcanobacterium pyogenes, Streptococcus spp., Corynebacterium spp., Actinobacillus spp., and Rhodococcus equi. The predominant identification of fecal and other opportunistic bacteria as opposed to pathogens commonly associated with equine abortion, such as Leptospira spp. and equine herpesvirus type 1 (EHV-1), suggests the need of improving hygiene management of breeding mares to prevent bacterial infection that may cause fetal loss, stillbirth, and perinatal mortality. Description: Abortion and complications of reproduction are important causes of economic losses in equine breeding due to the elevated costs incurred with diagnosis and treatment as well as the loss of animals (MOREIRA et al., 1998).

2020-01-01T00:00:00Z