Molecular detection and clinical aspects of porcine circovirus 3 infection in pigs from Brazil / [Detecção molecular e aspectos clínicos da infecção por circovírus suíno 3 em suínos do Brasil]

Porcine circovirus 3 (PCV-3) DNA has been detected in serum samples from apparently healthy pigs as well as pigs with different clinical conditions. Molecular detection of PCV-3 was observed in swine serum samples from Southeastern - Brazil using a nested PCR designed specifically for this study. The epidemiology and clinical aspects of PCV-3 infection were evaluated. The samples originated from 154 pigs of both genders from different production phases and with different clinical presentations, sampled from 31 pig farms visited between 2013 and 2018. In this study, PCV-3 was detected in 26.7% of samples from all populations across varying ages. Statistical association (P=0.0285) was observed only between animals with respiratory signs and PCV-3; no PCV-3-positive animal had diarrhea. No statistical association was observed between PCV-3 and age, or gender of the pigs. Because PCV-3 is a newly discovered virus, there is very little information about its epidemiology. We hope that these data can help in future studies investigating PCV-3 epidemiology.(AU)

O DNA do circovírus suíno 3 (PCV-3) foi detectado em amostras de soro de suínos aparentemente saudáveis, bem como em suínos com diferentes condições clínicas. A detecção molecular do PCV-3 foi observada em amostras de soro de suínos da região Sudeste do Brasil, com uma nested PCR desenhada especificamente para este estudo. A epidemiologia e os aspectos clínicos da infecção por PCV-3 foram avaliados. As amostras foram coletadas de 154 suínos de ambos os sexos, de diferentes fases de produção e com diferentes sinais clínicos. Os animais pertenciam a 31 granjas visitadas entre 2013 e 2018. Neste estudo, o PCV-3 foi detectado em 26,7% das amostras de animais saudáveis e de animais com variados sinais clínicos, de ambos os sexos e de idades variadas. Associação estatística (P=0,0285) foi observada apenas entre animais com sinais respiratórios e PCV-3; nenhum animal positivo para PCV-3 apresentava diarreia. Não foi observada associação estatística entre o PCV-3 e a idade ou o sexo dos suínos. Por se tratar de um vírus recém-descoberto, existem poucas informações sobre sua epidemiologia. Espera-se que os dados deste trabalho possam contribuir para futuros estudos sobre a epidemiologia do PCV-3.(AU)

Helicobacteriose em leitões: imuno-histoquímica em amostras colhidas por meio de gastroscopia / Helicobacter infection in piglets: Immunohistochemical analysis in mucosal samples collected using gastroscopy

O diagnóstico da úlcera gastroesofágica (UGE) é fundamental para que se tente a recuperação dos afetados, entretanto há grande dificuldade na sua realização de forma precisa. Modernos endoscópios são ferramentas importantes nesse sentido e podem ser usados para determinar em que fase da criação a ulceração ocorre. A bactéria Helicobacter tem sido associada à UGE e gerado perdas econômicas. Tendo em vista que poucos trabalhos têm sido realizados para identificação do problema precocemente, este estudo teve como objetivo detectar as lesões relacionando-as, ou não, com a bactéria Helicobacter spp., mediante análises macroscópica e histopatológica de amostras colhidas por meio da gastroscopia. Foram utilizados 20 animais de ambos os sexos, pesando entre 22 e 26kg e com 65 dias de idade. O aparelho utilizado foi um gastrovideoscópio da marca Karl Storz, modelo 1380NKS. Foram colhidas amostras das regiões anatômicas aglandular (quadrilátero esofágico) e glandular (cárdica, fúndica e pilórica) para o teste ultrarrápido da urease e para as avaliações histopatológicas e imuno-histoquímicas. Onze animais apresentaram lesões na região aglandular à endoscopia, e microscopicamente 15 animais apresentaram paraqueratose. Em 18 animais, foram observadas alterações em pelo menos uma das três regiões glandulares. As lesões foram maiores na região cárdica, seguida da antral e da fúndica. Em relação ao teste ultrarrápido da urease, sete animais foram negativos nas quatro regiões, e 13 positivos em pelo menos uma delas. Em relação à imuno-histoquímica (IHQ), 10 animais foram negativos em todas as regiões e 10 foram positivos em pelo menos uma delas. Os achados pré-ulcerativos não demonstraram relação com o Helicobacter spp., que apresenta caráter saprofítico e oportunista confirmado pela sua imunomarcação em áreas sem lesão...

Gastroesophageal ulcer (GEU) diagnosis is fundamental for the treatment and recovery of the affected animal stock. GEU is a condition affecting animals, resulting in depletion of animal stock and subsequent economic losses. Helicobacter spp. have been associated with GEU. Modern endoscopes are important for detecting the stage of the breeding process at which the ulceration occurs. However, few studies regarding early detection of GEU have been conducted. Therefore, we aimed to identify whether GEU lesions were related to Helicobacter spp. infection, using gastroscopy as a diagnostic technique for macroscopic and histopathological analyses. Twenty piglets (both male and female) with a mean age of 65 days were included (weight, 22-26 kg). We used a Karl Storz Gastrovideoscope (model 1380NKS). Samples from nonglandular and glandular (cardia, fundus, and pylorus) regions were collected for the ultra-rapid urease test and for histopathological and immunohistochemical (IHC) evaluations. Eleven animals showed macroscopic lesions in the nonglandular region during endoscopy, and 15 animals showed parakeratosis on histological analysis. Lesions in at least 1 glandular region were observed in 18 animals. The lesions were bigger in the cardiac region, followed by those in the antrum and the fundus. Regarding the ultra-rapid urease test, 7 animals were negative in all 4 regions and 13 were positive in at least 1 region. On IHC, 10 animals were negative in all 4 regions and 10 were positive in at least 1 region. However, pre-ulcerative findings were not correlated with Helicobacter spp. infection in the present study. The positive IHC findings for Helicobacter spp in regions without ulcerative lesions suggest its saprophytic and opportunistic nature...

Short-term and long-term antibody response by mice after immunization against Neisseria meningitidis B or diphtheria toxoid

Serogroup B Neisseria meningitidis (MenB) is a major cause of invasive disease in early childhood worldwide. The only MenB vaccine available in Brazil was produced in Cuba and has shown unsatisfactory efficacy when used to immunize millions of children in Brazil. In the present study, we compared the specific functional antibody responses evoked by the Cuban MenB vaccine with a standard vaccine against diphtheria (DTP: diphtheria, tetanus, pertussis) after primary immunization and boosting of mice. The peak of bactericidal and opsonic antibody titers to MenB and of neutralizing antibodies to diphtheria toxoid (DT) was reached after triple immunization with the MenB vaccine or DTP vaccine, respectively. However, 4 months after immunization, protective DT antibody levels were present in all DTP-vaccinated mice but in only 20% of the mice immunized against MenB. After 6 months of primary immunization, about 70% of animals still had protective neutralizing DT antibodies, but none had significant bactericidal antibodies to MenB. The booster doses of DTP or MenB vaccines produced a significant antibody recall response, suggesting that both vaccines were able to generate and maintain memory B cells during the period studied (6 months post-triple immunization). Therefore, due to the short duration of serological memory induced by the MenB vaccine (VA-MENGOC-BC® vaccine), its use should be restricted to outbreaks of meningococcal disease.

Investigação do gene p53 de frangos expostos às aflatoxinas / Investigation of p53 gene in poultries exposed to aflatoxins

Identificou-se o efeito das aflatoxinas (AFs) sobre o gene p53 de frangos de corte, de linhagem comercial, separados em: grupo experimental, tratado (GT) com ração comercial contendo 2,8ppm de AFs totais durante 21 dias consecutivos, e grupo-controle (GC), sem exposição às AFs. Macroscopicamente, as alterações caracterizaram-se por hepatomegalia e aspecto pálido-amarelado com alguns focos hemorrágicos e, histologicamente, por desarranjo trabecular, pleomorfismo hepatocítico com cariomegalia, degeneração vacuolar intracitoplasmática, necrose com infiltração linfocítica e hiperplasia de ductos biliares. A PCR com os primers GSPT53c-1 com base no gene candidato a p53 (GenBank XM_424937.2) gerou um produto de aproximadamente 350 pares de base. O amplicon sequenciado a partir do DNA dos frangos do GT não apresentou mutação ou deleção, assim como padrão de bandas do PCR-RFLP não foi distinto entre ambos os grupos experimentais e a sequência depositada no banco de genes. Os resultados sugerem que não ocorreu transversão devido à exposição às AFs no fragmento amplificado. Conclui-se que a PCR-RFLP e o sequenciamento do produto da PCR não são ferramentas apropriadas para diagnóstico da exposição de frangos às AFs nas condições experimentais empregadas.

To identify the effects of aflatoxins (AFs), Cobb lineage poultry were separated in an experimental group in which they were treated with commercial ration containing 2.8ppm of total AFs during 21 days (TG) and a control group without AFs exposure (CG). In the liver of poultries exposed to AFs, alterations were microscopically observed, which were characterized by hepatomegaly, a pale yellowish aspect with some hemorrhagic spots, and histologically a trabecullar disarranging pleomorphic hepatocytes with cariomegaly, intracytoplasmatic vacuolar degeneration, necrosis, lymphocytic infiltration and hyperplasia of biliary ducts. The PCR with GSPT53c-1 primers based on p53 candidate gen (GenBank XM_424937.2) generated a product of approximately 350 base pairs. The sequenced amplicon obtained from the DNA of treated poultry did not display any mutation or deletion, and the PCR- RFLP bands patterns were also not distinct in both experimental groups. The results indicated that transversion did not occur in the fragment amplified due to AFs exposure. As a consequence of results obtained with p53 gene (NM_205264.1) we concluded that PCR-RFLP and sequencing of PCR product are not appropriate diagnostic tools for the detection of poultry exposure to AFs, at least in the experimental conditions performed.

Demodicose suína no norte do estado do Rio de Janeiro: relato de caso / Demodicosis in northern Rio de Janeiro State: case report

Um leitão oriundo de um criatório do norte do estado do Rio de Janeiro, mantido no biotério do Hospital Veterinário da Universidade Estadual do Norte Fluminense Darcy Ribeiro e utilizado como caso piloto de projeto de pesquisa para investigações sobre o acesso videolaparoscópico retal (NOTES - Natural Orifice Translumenal Endoscopic Surgery), foi eutanasiado e necropsiado. Amostras do ponto de acesso do tubo de inserção na região mucocutânea anal foram colhidas, fixadas em formalina neutra tamponada a 10%, processadas por inclusão em parafina e coradas pela hematoxilina e eosina para histologia. Observou-se o comprometimento de folículos que, dilatados, exibiam formações que lembravam Demodex spp. cortados em vários sentidos e, por se tratar de ácaros espécie-específicos, concluiu-se como D. phylloides. Na citopatologia, o ácaro foi recuperado e identificado a fresco, mostrando-se alongado, com quatro pares de patas ventrais e no terço anterior do corpo. Este se refere ao primeiro registro de demodicose suína no estado do Rio de Janeiro.

A piglet derived from a farm in northern Rio de Janeiro State, maintained at the Animal Veterinary Hospital of Universidade Estadual do Norte Fluminense Darcy Ribeiro and used as a pilot case for a research project on access to research video-laparoscopic rectal (NOTES - Natural Orifice Endoscopic Surgery Translumenal) was euthanized and necropsied. Samples from the access point of the insertion tube in mucocutaneous anal region were collected, fixed in buffered formalin 10%, processed by embedding in paraffin and stained with hematoxylin and eosin for histopathology. The impairment of follicles that exhibited dilated formations resembling Demodex spp. cut in various directions and due to the mite species-specific was observed, and was concluded as D. phylloides. The mite was recovered and freshly identified by cytopathology. It was elongated, with four pairs of legs, at ventral and anterior third of the body. This refers to the first report of swine demodicosis in Rio de Janeiro State.

Flaxseed energy and macronutrients balance.

BACKGROUND/OBJECTIVES: Flaxseed has functional properties in the reduction of the risk of chronic non-communicable diseases such as cardiovascular disease, diabetes and cancer. Regardless of its high energy density, the consumption of flaxseed tends to promote body weight maintenance. The purpose of this study was to evaluate energy and macronutrient balance after flaxseed consumption. SUBJECTS/METHODS: Twenty four healthy volunteers were allocated into 3 experimental groups, when they consumed flaxseed (FS), defatted flaxseed flour (FF), or flaxseed oil (FO). During the control period they were provided a diet without flaxseed products for 7-9 days. Following that diets containing 70 g of one of the flaxseed products were consumed for another 7-9 day- period. Test foods were consumed exclusively in the laboratory and fecal excretion was collected during the study. There was a higher energy excretion (P < 0.05) in the FF and FS groups, compared to their control and FO group. RESULTS: The excretions of total lipid and the PUFA α-linolenic acid were higher in FS group (P < 0.05). CONCLUSIONS: The intake of 70 g/day of FS and FF raised lipid and energy excretion, which may mitigated the effect of flaxseed consumption on body weight.

Specific elimination of effector memory CD4+ T cells due to enhanced Fas signaling complex formation and association with lipid raft microdomains.

Elimination of autoreactive CD4(+) T cells through the death receptor Fas/CD95 is an important mechanism of immunological self-tolerance. Fas deficiency results in systemic autoimmunity, yet does not affect the kinetics of T-cell responses to acute antigen exposure or infection. Here we show that Fas and TCR-induced apoptosis are largely restricted to CD4(+) T cells with an effector memory phenotype (effector memory T cells (T(EM))), whereas central memory and activated naïve CD4(+) T cells are relatively resistant to both. Sensitivity of T(EM) to Fas-induced apoptosis depends on enrichment of Fas in lipid raft microdomains, and is linked to more efficient formation of the Fas death-inducing signaling complex. These results explain how Fas can cull T cells reactive against self-antigens without affecting acute immune responses. This work also identifies Fas-induced apoptosis as a possible immunotherapeutic strategy to eliminate T(EM) linked to the pathogenesis of a number of autoimmune diseases.

Heterogeneity in envelope protein sequence and N-linked glycosylation among yellow fever virus vaccine strains.

We have compared the deduced envelope (E) protein sequences of two biologically well-characterized yellow fever (YF) virus vaccine strains. The 17DD strain has been produced in Brazil for more than 50 years and used to successfully vaccinate millions of people worldwide. The 17D-213 is a candidate vaccine strain produced in tissue culture which has previously passed the monkey neurovirulence assay for testing human YF vaccines. Nucleotide sequence analysis of polymerase chain reaction-amplified cDNA revealed a number of mutations which were strain- and substrain-specific. A major difference of 17DD and 17D-213 as compared to 17D-204 and Asibi was the existence of a potential N-linked glycosylation site located at amino acid residues 153 and 151 of 17DD and 17D-213, respectively. These acceptor sites are apparently utilized for the addition of high-mannose carbohydrate chains as shown by endoglycosidase analyses of immunoprecipitated E proteins. Glycosylated E protein is also used to assemble YF vaccine virions. This work and eventual complete nucleotide sequence analysis of both vaccine strains should help to define possible changes involved in YF virus attenuation and allow their biological importance to be determined using a recently developed system for generating YF virus from cDNA. In addition, these data provide an estimate on the extent of genetic variability among YF 17D seeds and vaccines.

Treatment of human taeniasis in the Philippines: a review.

Studies relating to the treatment of human taeniasis in the Philippines using Atabrine, bithionol, and mebendazole are reviewed. Cure rates of 93.7% for Atabrine, 85%-100% for bithionol, and 84.62% for mebendazole are reported. Aside from these studies, recent experience in the treatment of Taenia saginata infections at the Department of Parasitology, College of Public Health, is also reported. Twenty patients referred to the department were treated, 3 with bithionol, 17 with praziquantel. Of the 17 patients treated with praziquantel, the study reported cure rates of 47-88.24%.

Hepatitis B: a case for prenatal screening of all patients.

Evaluation of a mass screening program to detect hepatitis B surface antigen in the obstetric population of Shands Hospital, University of Florida in Gainesville, from January 1, 1983, through December 31, 1985, was undertaken. Prevalence of hepatitis B surface antigen seropositivity was 0.54%. Review of medical records revealed that 67% of patients with positive hepatitis B surface antigen screens had no risk factors identifiable by routine prenatal history. Although all patients were screened on admission to the hospital, the results of the immunoassay were not available in time for staff to institute isolation procedures for the laboring mother or unwashed neonate. In 82% of the cases, treatment of neonates occurred later than 12 hours after delivery (the current Centers for Disease Control recommendation). It is recommended that prenatal screening of all patients with a hepatitis B surface antigen immunoassay be done by 34 weeks' gestation where the patient population is of predominantly low socioeconomic status to ensure appropriate isolation and timely neonatal immunoprophylaxis.

Sonographic detection of fetal urinary-tract anomalies.

The diagnosis of urinary-tract anomalies in the fetus can frequently be made with antenatal ultrasound imaging. This report details 19 cases of fetal urinary-tract malformation diagnosed during a 30-month period. Early onset and more complete urinary-tract obstruction, oligohydramnios, renal dysplasia, and pulmonary hypoplasia occur frequently in cases where urinary-tract anomalies were detected in utero. The frequent association of other major structural anomalies, severe renal damage, and irreversible pulmonary hypoplasia make imperative an intensive evaluation of the fetus before considering invasive surgical procedures.

Immunocytochemical localization of relaxin in human decidua and placenta.

Specimens from 20 human term placentas were stained with 4 different antisera produced against porcine relaxin (Rlx) using the avidin-biotin immunoperoxidase procedure. Cells of the parietal decidua adherent to the fetal membranes, cells of the chorionic cytotrophoblast, as well as cells of the placental basal plate consistently stained with all 4 anti-Rlx sera. Occasionally, Rlx was detected in epithelial cells lining the amniotic membrane. The syncytiotrophoblast stained for Rlx in 2 specimens only. This response was seen only in syncytiotrophoblast that lined villi in close proximity to the basal plate. Syncytiotrophoblast of the chorionic villi either did not stain at all or gave very weak positive immunostaining with the anti-Rlx sera in all specimens. No difference was noted in staining patterns among placentas delivered by elective cesarean section or vaginal delivery.

Reproductive failure and cervical incompetence in women exposed to diethylstilbestrol.

Exposure to diethylstilbestrol in utero is recognized to be associated with a wide range of developmental abnormalities in the female genital tract, the most prominent being adenocarcinoma of the vagina. Recently anatomic anomalies have been discovered with increasing frequency. This case illustrates the association of exposure to diethylstilbestrol, cervical incompetence, and poor reproductive history, and stresses the importance of early evaluation.