Metabolic phenotype modulation by caloric restriction in a lifelong dog study

Modeling aging and age-related pathologies presents a substantial analytical challenge given the complexity of gene−environment influences and interactions operating on an individual. A top-down systems approach is used to model the effects of lifelong caloric restriction, which is known to extend life span in several animal models. The metabolic phenotypes of caloric-restricted (CR; n = 24) and pair-housed control-fed (CF; n = 24) Labrador Retriever dogs were investigated by use of orthogonal projection to latent structures discriminant analysis (OPLS-DA) to model both generic and age-specific responses to caloric restriction from the 1H NMR blood serum profiles of young and older dogs. Three aging metabolic phenotypes were resolved: (i) an aging metabolic phenotype independent of diet, characterized by high levels of glutamine, creatinine, methylamine, dimethylamine, trimethylamine N-oxide, and glycerophosphocholine and decreasing levels of glycine, aspartate, creatine and citrate indicative of metabolic changes associated largely with muscle mass; (ii) an aging metabolic phenotype specific to CR dogs that consisted of relatively lower levels of glucose, acetate, choline, and tyrosine and relatively higher serum levels of phosphocholine with increased age in the CR population; (iii) an aging metabolic phenotype specific to CF dogs including lower levels of liproprotein fatty acyl groups and allantoin and relatively higher levels of formate with increased age in the CF population. There was no diet metabotype that consistently differentiated the CF and CR dogs irrespective of age. Glucose consistently discriminated between feeding regimes in dogs (≥312 weeks), being relatively lower in the CR group. However, it was observed that creatine and amino acids (valine, leucine, isoleucine, lysine, and phenylalanine) were lower in the CR dogs (<312 weeks), suggestive of differences in energy source utilization. 1H NMR spectroscopic analysis of longitudinal serum profiles enabled an unbiased evaluation of the metabolic markers modulated by a lifetime of caloric restriction and showed differences in the metabolic phenotype of aging due to caloric restriction, which contributes to longevity studies in caloric-restricted animals. Furthermore, OPLS-DA provided a framework such that significant metabolites relating to life extension could be differentiated and integrated with aging processes.

Multiplexed femtomolar quantitation of human cytokines in a fluoropolymer microcapillary film

Sensitive quantitation of multiple cytokines can provide important diagnostic information during infection, inflammation and immunopathology. In this study sensitive immunoassay detection of human cytokines IL-1β, IL-6, IL-12p70 and TNFα is shown for singleplex and multiplex formats using a novel miniaturized ELISA platform. The platform uses a disposable plastic multi-syringe aspirator (MSA) integrating 8 disposable fluoropolymer microfluidic test strips, each containing an array of ten 200 mean i.d. microcapillaries coated with a set of monoclonal antibodies. Each MSA device thus performs 10 tests on 8 samples, delivering 80 measurements. Unprecedented levels of sensitivity were obtained with the novel fluoropolymer microfluidic material and simple colorimetric detection in a flatbed scanner. The limit of detection for singleplex detection ranged from 2.0 to 15.0 pg/ml, i.e. 35 and 713 femtomolar for singleplex cytokine detection, and the intra- and inter-assay coefficient of variation (CV) remained within 10%. In addition, a triplex immunoassay was developed for measuring IL-1β, IL-12p70 and TNFα simultaneously from a given sample in the pg/ml range. These assays permit high sensitivity measurement with rapid <15 min assay or detection from undiluted blood serum. The portability, speed and low-cost of this system are highly suited to point-of-care testing and field diagnostics applications.

Search for Salmonella spp. in ostrich productive chain of Brazilian southeast region

We analyzed ostriches from an equipped farm located in the Brazilian southeast region for the presence of Salmonella spp. This bacterium was investigated in 80 samples of ostrich droppings, 90 eggs, 30 samples of feed and 30 samples of droppings from rodents. Additionally, at slaughter-house this bacterium was investigated in droppings, caecal content, spleen, liver and carcasses from 90 slaughtered ostriches from the studied farm. Also, blood serum of those animals were harvested and submitted to serum plate agglutination using commercial Salmonella Pullorum antigen. No Salmonella spp. was detected in any eggs, caecal content, liver, spleen, carcass and droppings from ostriches and rodents. However, Salmonella Javiana and Salmonella enterica subsp. enterica 4, 12: i:- were isolated from some samples of feed. The serologic test was negative for all samples. Good sanitary farming management and the application of HACCP principles and GMP during the slaughtering process could explain the absence of Salmonella spp. in the tested samples.

Differential temporal expression of milk miRNA during the lactation cycle of the marsupial tammar wallaby (Macropus eugenii)

Lactation is a key aspect of mammalian evolution for adaptation of various reproductive strategies along different mammalian lineages. Marsupials, such as tammar wallaby, adopted a short gestation and a relatively long lactation cycle, the newborn is immature at birth and significant development occurs postnatally during lactation. Continuous changes of tammar milk composition may contribute to development and immune protection of pouch young. Here, in order to address the putative contribution of newly identified secretory milk miRNA in these processes, high throughput sequencing of miRNAs collected from tammar milk at different time points of lactation was conducted. A comparative analysis was performed to find distribution of miRNA in milk and blood serum of lactating wallaby.

Foot-and-mouth disease in red deer - experimental infection and test methods performance

Summary: The aim of this study was to evaluate a number of foot-and-mouth disease (FMD) test methods for use in red deer. Ten animals were intranasally inoculated with the FMD virus (FMDV) O UKG 11/2001, monitored for clinical signs, and samples taken regularly (blood, serum, oral swabs, nasal swabs, probang samples and lesion swabs, if present) over a 4-week period. Only one animal, deer 1103, developed clinical signs (lesions under the tongue and at the coronary band of the right hind hoof). It tested positive by 3D and IRES real-time reverse transcription polymerase chain reaction (rRT-PCR) in various swabs, lesion materials and serum. In a non-structural protein (NSP) in-house ELISA (NSP-ELISA-IH), one commercial ELISA (NSP-ELISA-PR) and a commercial antibody NSP pen side test, only deer 1103 showed positive results from day post-inoculation (dpi) 14 onwards. Two other NSP-ELISAs detected anti-NSP serum antibodies with lower sensitivity. It also showed rising antibody levels in the virus neutralization test (VNT), the in-house SPO-ELISA-IH and the commercial SPO-ELISA-PR at dpi 9, and in another two commercial SPO-ELISAs at dpi 12 (SPO-ELISA-IV) and dpi 19 (SPO-ELISA-IZ), respectively. Six of the red deer that had been rRT-PCR and antibody negative were re-inoculated intramuscularly with the same O-serotype FMDV at dpi 14. None of these animals became rRT-PCR or NSP-ELISA positive, but all six animals became positive in the VNT, the in-house SPO-ELISA-IH and the commercial SPO-ELISA-PR. Two other commercial SPO-ELISAs were less sensitive or failed to detect animals as positive. The rRT-PCRs and the four most sensitive commercial ELISAs that had been used for the experimentally inoculated deer were further evaluated for diagnostic specificity (DSP) using 950 serum samples and 200 nasal swabs from non-infected animals. DSPs were 100% for the rRT-PCRs and between 99.8 and 100% for the ELISAs.

Perdas endógenas e coeficientes de absorção aparente e real do magnésio em caprinos

Os objetivos deste trabalho foram determinar a absorção aparente, estimar as perdas endógenas fecais e a absorção real do Mg e determinar a ingestão ad libitum da água de beber e a concentração de Mg no soro sangüíneo de caprinos da raças Anglonubiana (AN) e Saanen (SN). Foram usados doze caprinos, seis de cada raça, com 19,8 kg PV médio. Dietas semipurificadas (baixo teor de Mg) à base de quirera de arroz, glúten de milho e celulose foram suplementadas com MgO, para se obterem os níveis de 0,05 (sem supplementação) 0,20 e 0,35% Mg (%MS). Os níveis de Mg influenciaram os coeficientes de absorção aparente de Mg e Ca, com valores médios de 57,8; 73,9; e 73,2% para Mg e 55,7; 39,6; e 49,5% para Ca, para dietas com níveis 0,05; 0,20; e 0,35% de Mg, respectivamente. Entretanto, para os coeficientes de absorção aparente de P, Na e K, não houve efeito de níveis de Mg na dieta. Os resultados de absorção real de Mg apresentaram interação de níveis de Mg e raças. A média para raça NA, no nível 0,05% Mg, foi de 61,0% e para os níveis 0,20 e 0,35% Mg, 77,2 e 73,2%, respectivamente. Entretanto, para a raça SN, as médias foram 73,3; 75,5; e 76,0%, para os mesmos níveis, sem diferenças. A digestibilidade de matéria seca, proteína bruta e extrato não-nitrogenado diminuiu com os níveis crescentes de Mg nas dietas. As excreções fecais (7,0; 20,8; e 34,4 mg/kg PV0,75.d) e urinárias (3,9; 30,8; e 44,6 mg/kg PV0,75.d) de Mg elevaram-se com o aumento dos níveis crescentes de Mg nas dietas. Houve, também, influência dos níveis de Mg dietético sobre as concentrações de Mg do soro sangüíneo (1,74; 2,23; e 2,80 mg/dL para níveis de 0,05; 0,20; e 0,35% de Mg, respectivamente).

Proteinograma sérico de cães sadios e com linfoma obtido por eletroforese em gel de poliacrilamida (SDS-PAGE)

Foram avaliadas amostras de soro sanguíneo de 10 cães sadios e de 12 com linfoma, utilizando-se a eletroforese em gel de poliacrilamida contendo dodecil sulfato de sódio. Houve diferença entre as médias dos teores de proteína total de cães sadios, 7,68g/dL±0,46 e de cães com linfoma, 7,93g/dL±2,49. As concentrações de IgA e IgG não foram diferentes entre os grupos. Os teores das proteínas de pesos moleculares 142000, 110000, 52000, 49000, 24000 e 18000 dáltons foram mais elevados em cães com linfoma. Os cães com linfoma apresentaram concentrações mais elevadas de ceruloplasmina, 43,95mg/dL±18,19, e haptoglobina, 554mg/dL±449,51, e menores de albumina, 2908mg/dL±476,67, em comparação aos cães sadios (ceruloplasmina: 3,42mg/dL±7,44; haptoglobina: 94,54mg/dL±59,50 e albumina: 4207mg/dL±206,18). Conclui-se que concentrações séricas mais elevadas de ceruloplasmina e haptoglobina e menores de albumina podem estar associadas ao linfoma em cães.

Influência do puerpério sobre o proteinograma sérico de caprinos da raça Saanen obtido por eletroforese em gel de poliacrilamida

Aiming to evaluate the puerperal influence on the proteinogram of Saanen goats, 108 samples of blood serum from 12 goats were collected, and the results were presented at nine times: just after parturition, 1, 3, 5, 7, 10, 15, 21 and 30 days after parturition. Total amount of serum proteins were determined by the biuret technique, and the sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to the protein fractionation. In this last method, 17 protein bands were observed, from which molecular weights varied between 25 KDa and 275 KDa. In addition, it was possible to identify the following protein fractions: immunoglobulin A (180 KDa), ceruloplasmin (115 KDa), transferrin (79 KDa), albumin (65 KDa), heavy-chain immunoglobulin G (58 KDa), haptoglobin (45 KDa), acid glycoprotein (37 KDa) and light-chain immunoglobulin G (28 KDa). Another 9 nonidentified protein fractions presented, each molecular weights equal to 275 KDa, 140 KDa, 125 KDa, 103 KDa, 95 KDa, 41 KDa, 35 KDa, 30 Kda and 25 KDa. The results allow us to conclude that by the first week of puerperium, an improvement of acid glycoprotein occurs, whereas those others protein fractions do not suffer any puerperal influence.

Search for Salmonella spp. in ostrich productive chain of Brazilian southeast region

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

Epidemiological and Serological Aspects in Canine Toxoplasmosis in Animals with Nervous Symptoms

The presence of anti-Toxoplasma gondii IgM and IgG antibodies was studied in samples of blood serum taken from eighty dogs with nervous symptoms at the Serviço de Enfermidades Infecciosas dos Animais, Faculdade de Medicina Veterinária e Zootecnia, Unesp, Botucatu, São Paulo, Brazil. The frequency of IgG titers were 16 (13.7%), 64 (13.7%), and 256 (5%), and for IgM titers were 16 (7.5%), 64 (15%), and 256 (8.7%). Positive reactions were more frequent in the older animals, males, from a rural environment, in constant contact with small animals, principally birds and rodents. There was a higher frequency of a positive reaction in dogs fed with kitchen food, especially in those fed with raw ingredients. The most common neurological pictures were alterations in consciousness, in movement, and in the hand-cart test. The percentage of reagents with specific IgM antibodies was high, indicating active infections, but the possibility of co-infection with the distemper virus can not be discarded, and this may be a predisposing factor for toxoplasmosis infection, once the distemper virus has a potent immunosupressive action.

Anticorpos contra Toxoplasma gondii em equinos com histórico de ataxia

Stelmann U.J.P., Silva R.C., Langoni H., Borges A.S. & Amorim R.M. [Antibodies against Toxoplasma gondii in horses with history of ataxia]. Anticorpos contra Toxoplasma gondii em equinos corn historic de ataxia. Revista Brasileira de Medicina Veterinaria, 33(4):200-202, 2011. Programa de Pos-Graduacao em Medicina Veterinaria, Departamento de Clinica Veterinaria, Faculdade de Medicina Veterinaria e Zootecnia, Universidade Estadual Paulista Julio de Mesquita Filho, Campus de Botucatu, Distrito de Rubiao Junior, s/n, Botucatu, SP 18618-970, Brasil. E-mail: stelmann.ppgctia@gmail.comThe frequency of antibodies in blood serum and cerebrospinal fluid against Toxoplasma gondii in horses with a history of ataxia from the state of São Paulo, Brazil was carried out. Modified agglutination test was used to determine antibodies against T gondii, considering as positive samples with titers >= 2. of the blood samples tested, only 8 of 23 (34.78%) were positive, while CSF samples were negative when used the same technique. According to the negative results obtained for CSF, we concluded that T gondii was not the etiological agent of the myeloencephalitis in studied horses with neurological manifestation of ataxia, while the blood serological results indicated a previously exposure to T gondii.

Animal cysticercosis in indigenous Brazilian villages

A inspeção sanitária da carne bovina e suína tem sido a principal forma diagnóstica da cisticercose animal e da prevenção da teníase no Brasil. As aldeias indígenas Jaguapirú e Bororó estão localizadas próximo à área urbana do município de Dourados-MS, com condições precárias de saneamento básico, onde bovinos e suínos são criados como fonte de alimento para consumo próprio, bem como para comercialização externa, geralmente sem inspeção sanitária oficial. Neste estudo, 96 carcaças bovinas e 117 amostras de soro de suínos, criados nas aldeias indígenas, foram avaliadas para a presença de formas metacestóides à inspeção sanitária e de anticorpos anti-Taenia sp. ao teste ELISA, respectivamente. Observaram-se 18,75% de positividade para cisticercose bovina e 9,4% para cisticercose suína. A ocorrência do complexo teníase-cisticercose nas aldeias pode favorecer a ocorrência desta zoonose na população indígena. Condições adequadas de abate e inspeção sanitária dos animais destas aldeias se fazem urgente para o controle do complexo teníase-cisticercose na população indígena.

Naturally acquired antibodies against Clostridium perfringens epsilon toxin in goats

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

Normalização dos níveis séricos de ácido ascórbico por suplementação com suco de acerola (Malpighia glabra L.) ou farmacológica em idosos institucionalizados

INTRODUÇÃO: Este estudo investigou o tempo necessário de suplementação com vitamina C, para a normalização dos níveis séricos em idosos com deficiência dessa vitamina e comparar o efeito da vitamina natural do suco de acerola (Malpighia glabra L.) com o da vitamina na forma de fármaco. MÉTODOS: Foram estudados 37 idosos institucionalizados do município de João Pessoa, Paraíba, Brasil, divididos em 3 grupos: Grupo I - controle, Grupo II - suplementação com o suco de acerola e Grupo III - suplementação com fármaco. A metodologia empregada consistiu na dosagem sérica de ácido ascórbico e na verificação do consumo alimentar por inquérito dietético. Constatou-se um aumento significativo (p<0,05) nas médias dos níveis séricos de ácido ascórbico, após 10 dias (1,27±0,41mg/dL), 20 (1,69±0,45mg/dL) e 30 dias (1,55±0,42mg/dL) de suplementação aos valores iniciais (0,38±0,28mg/dL). No 10º dia de suplementação, os idosos suplementados com suco de acerola apresentaram níveis significativamente mais elevados (1,41±0,43mg/dL) do que aqueles que foram suplementados com comprimidos (1,03±0,25mg/dL). CONCLUSÃO: Considerando-se que, no 20º dia, o efeito da suplementação foi satisfatório para a normalização dos níveis séricos daqueles indivíduos, esse tempo poderia ser utilizado para idosos em geral e, em especial, para aqueles que vivem em instituições destinadas a idosos carentes, sendo o suco de acerola um suplemento indicado por ser um produto natural e de fácil aquisição.

Comparative cytogenetic studies of Bufo ictericus, B. paracnemis (Amphibia, Anura) and an intermediate form in sympatry

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