Establishment of a production process for a novel vaccine candidate against Lawsonia intracellularis

BACKGROUND: Lawsonia intracellularis remains a problem for the swine industry worldwide. Previously, we designed and obtained a vaccine candidate against this pathogen based on the chimeric proteins: OMP1c, OMP2c, and INVASc. These proteins formed inclusion bodies when expressed in E. coli, which induced humoral and cellular immune responses in vaccinated pigs. Also, protection was demonstrated after the challenge. In this study, we established a production process to increase the yields of the three antigens as a vaccine candidate. RESULTS: Batch and fed-batch fermentations were evaluated in different culture conditions using a 2 L bioreactor. A fed-batch culture with a modified Terrific broth medium containing glucose instead of glycerol, and induced with 0.75 mM IPTG at 8 h of culture (11 g/L of biomass) raised the volumetric yield to 627.1 mg/L. Under these culture conditions, plasmid-bearing cells increased by 10% at the induction time. High efficiency in cell disruption was obtained at passage six using a high-pressure homogenizer and a bead mill. The total antigen recovery was 64% (400 mg/L), with a purity degree of 70%. The antigens retained their immunogenicity in pigs, inducing high antibody titers. CONCLUSIONS: Considering that the antigen production process allowed an increment of more than 70-fold, this methodology constitutes a crucial step in the production of this vaccine candidate against L. intracellularis.

Período de adaptação a dietas com baixa ou alta fibra sobre a digestibilidade e as características fecais em cães / Adaptation period to diets with low or high fiber on digetibility and faecal characteristics in dogs

Objetivou-se avaliar três períodos de adaptação, cinco, 10 e 15 dias, a dietas com baixa ou alta fibra sobre a digestibilidade e características fecais. Foram utilizadas duas dietas, uma contendo 0% de casca de soja (0%CS) e outra com 15% de casca de soja (15%CS), fornecidas a 12 cães adultos. Ao final de cada período, houve coleta total de fezes para avaliação dos coeficientes de digestibilidade aparente (CDA), energia metabolizável (EM) e características fecais. Além disso, foi avaliada a palatabilidade das dietas. Como resultado, não houve efeito do período de adaptação sobre a digestibilidade das dietas (P>0,05). A dieta 15%CS apresentou menor CDA da matéria seca (MS) e EM, em relação à dieta 0%CS (P<0,05). A consistência fecal dos cães não diferiu entre as dietas e os períodos (P>0,05). Houve redução no pH e na amônia fecal dos cães alimentados com a dieta 15%CS (P<0,05), após 10 dias de alimentação. Não houve diferença na palatabilidade das dietas (P>0,05). Períodos de adaptação à dieta superiores a cinco dias não alteram a digestibilidade e a maioria das características fecais de cães, exceto a amônia fecal. A inclusão de 15%CS na dieta reduz a digestibilidade da MS e a EM, bem como o pH e a amônia fecal.(AU)

This study aimed to evaluate three periods of adaptation, 5, 10 and 15 days, to diets with low or high fiber on digestibility and fecal characteristics. Two diets were used, one containing 0% (0% SH) and one containing 15% soybean hulls (15% SH). Diets were provided to 12 adult dogs. At the end of each period there was total collection of feces to evaluate the apparent digestibility coefficients (ADC), metabolizable energy (ME) and fecal characteristics. We also evaluated diet palatability. There was no effect of the adaptation period on digestibility of diets (P> 0.05). The 15% SH diet reduced ADC of dry matter (DM) and ME, compared to the 0% SH diet (P< 0.05). Fecal consistency of the dogs did not differ among diets and periods (P> 0.05). There was a decrease in fecal pH and ammonia in dogs fed the diet with 15% SH (P< 0.05) after 10 days of feeding (P< 0.05). There was no difference in diet palatability (P> 0.05). Diet adaptation longer than 5 days did not alter the digestibility and most fecal characteristics of dogs, except for fecal ammonia. The inclusion of 15% SH in diet reduces DM digestibility and fecal pH and ammonia in dogs.(AU)

Requirements and energy efficiency of Pelibuey and Katahdin non pregnant, non lactating ewes in Yucatan, Mexico / Requerimientos y eficiencia energética de ovejas Pelibuey y Katahdin no gestantes, no lactantes en Yucatán, México

Abstract Objective. To estimate metabolizable energy requirements for maintenance (MEm), energy efficiency of weight gain (EEWG) and fat thickness, in Pelibuey and Katahdin ewes in Yucatan, Mexico. Material and method. Eight non pregnant, non lactating, multiparous ewes, were feed at three intake levels with a diet containing 2.0 Mcal/Kg-1 of ME and 11 % of CP. Feed intake was measured daily, ewes weight and thickness of subcutaneous fat (SF) were determined every 14 days. The EMm was estimated by regressing values of live weight against metabolizable energy intake (MEI), EEWG was estimate as gram of weight gain per Mcal of MEI. Results. There were no differences between breeds in MEm (97±4 and 110±4 kcal/Kg0.75, for Pelibuey and Katahdin) and EEWG (58±8 and 63±8 g/Mcal of MEI, for Pelibuey and Katahdin); differences were found in SF (6.1±0.2 and 4.9±0.2 mm, for Pelibuey and Katahdin). Conclusions. The energy requirements for maintenance were similar in Pelibuey and Katahdin ewes in Yucatan, Mexico.

Resumen Objetivo. Estimar los requerimientos de energía metabolizable para el mantenimiento (MEm), le eficiencia energética del incremento de peso (EEWG) y el grosor de grasa, en ovejas Pelibuey y Katahdin en Yucatán, México. Materiales y método. Ocho ovejas multíparas no gestantes, no lactantes, fueron alimentadas a tres niveles de consumo con una dieta que contenía 2.0 Mcal/kg-1 de EM y 11% de PC. El consumo de alimento fue medido diariamente, el peso de ovejas y el grosor de grasa subcutánea (SF) se determinó cada 14 días. La MEm fue estimada por regresión de los valores de peso vivo contra el consumo de EM (MEI), la EEWG fue estimada como los gramos de ganancia de peso por Mcal de MEI. Resultados. No existieron diferencias entre razas en EMm (97±4 y 110±4 kcal/Kg0.75, para Pelibuey y Katahdin respectivamente) y EEWG (58±8 y 63±8 g/Mcal de MEI, para Pelibuey y Katahdin respectivamente); se encontraron diferencias en SF (6.1±0.2 y 4.9±0.2 mm, para Pelibuey y Katahdin respectivamente). Conclusiones. Los requerimientos de energía para mantenimiento fueron similares en las ovejas Pelibuey y Katahdin en Yucatán, México.

Probable enfermedad de Kawasaki diagnosticada en fase de secuelas en un hombre adulto de 56 años de edad / Possible Kawasaki disease in a 56 years-old male patient diagnosed in sequelae phase

Se presenta el caso de un paciente de 56 años edad que sufre un evento isquémico agudo con arterias coronarias con una importante dilatación en los segmentos proximales izquierdos y una dilatación de hasta 10 mm de diámetro en la coronaria derecha proximal y todo el segmento vertical, compatibles con enfermedad de Kawasaki en fase de secuelas crónicas.

We present a 56 years-old male patient with an acute ischemic episode. The patient showed significant dilation of the left main coronary artery and proximal segments of the left descending artery, as well as a right coronary artery with a 10 mm diameter in the proximal and medial segments. These findings are compatible with a Kawasaki's disease in a chronic phase.

Use of Triticum aestivum in open wound healing: a clinical, pathological, and tensiometric assessment in the rabbit model / Uso de Triticum aestivum na cicatrização de feridas: um ensaio clínico, histopatológico e tensiométrico em coelhos

While Triticum sp. has been shown to act in wound healing, stimulating collagen synthesis by fibroblasts, the use of this plant extract has yet to be assessed in vivo, in commercially viable presentations. This study used rabbits and assessed, on days seven, 14, and 21, the presence or absence of granulation tissue and epithelialization, histopathological structures, and scar quality through the breaking and tension strength. Treatments, performed for 21 days, were aqueous extract of T. aestivum at a concentration of 2mg/mL (group I) and 10mg/mL (group II) and a nonionic cream (control group). We demonstrate that the formation of granulation tissue was not significantly different between treatments. In the analysis of epithelial tissue, wounds in group II differed from other treatments by day 7. On days 14 and 21 there was no significant clinical difference between groups. In the histopathological evaluation, scar quality and rupture strength did not differ between the groups in the studied period. In the tension strength evaluation, group I differed from the others, presenting a higher tension strength overall. The studied treatments did not differ regarding healing evolution of the skin wounds, but T. aestivum extract, at 2mg/mL, presents better results in the tension strength evaluation...

O extrato de trigo (Triticum sp.) vem sendo usado na cicatrização de feridas por estimular a síntese de fibroblastos, entretanto a sua aplicabilidade in vivo em apresentações comercialmente viáveis ainda tem de ser demonstrada. Neste estudo, avaliaram-se feridas cutâneas de coelhos tratadas com extrato aquoso de T. aestivum quanto à presença de tecido de granulação e epitelização, estruturas histológicas, qualidade cicatricial, além de ensaio tensiométrico. As feridas foram tratadas diariamente, por 21 dias, com diferentes concentrações do extrato (grupo I = 2mg/mL; grupo II = 10mg/mL) ou apenas o veículo (grupo controle = creme não iônico), e avaliadas nos dias sete, 14 e 21. A formação de tecido de granulação não diferiu entre os tratamentos. A epitelização aconteceu em menor tempo em feridas do grupo II, mas aos 14 dias já não havia diferença neste parâmetro. Na avaliação histopatológica, a qualidade cicatricial e a força de ruptura não diferiram no período estudado, entretanto a resistência tensiométrica das feridas do grupo I foi maior que a dos demais tratamentos. Dessa forma, conclui-se que, mesmo não havendo diferença na evolução cicatricial de feridas tratadas ou não com extrato aquoso de T. aestivum, o uso desse composto, a 2mg/mL, resultou em tecidos cicatriciais mais resistentes à tração...

Infection in immune-deficiency states: aplastic anemia and leukemia a two-year review of cases at the UP-PGH Medical Center, (1974-1975)

In this two-year review of 107 cases of Aplastic Anemia, and Acute and Chronic Leukemia, it was found that the focus of infection in over one half of the total number of cases, was pulmonary, with soft tissue and urinary tract infection ranking second and third. The Onset of infection was found to be approximately 15 days after admission in aplastics, and 12 days for leukemicsStaphylococcus Aureaus was the most common pathogen in community acquired infections, while Pseudomonas, Klebsiella and E. Coli were common among the hospital acquired group. More deaths were associated with gram-negative organisms and, at a mean neutrophile count of 1,450/cu.mm. However, no significant relationship could be established between death and lymphocyte levelIn leukemia, markedly low neutrophile counts were observed at the time infection was known to set in, while no change was observed in the aplastic group, from admission to time of infection. It was only with the chronic leukemics that a significant decrease in lymphocyte count was noted. (Summary)

Esclerose tuberosa: estudo multidisciplinar de 15 casos / Tuberose esclerosis: a multidisciplinary study of 15 cases

Sao apresentados os resultados de estudo multidisciplinar de 15 pacientes con esclerose tuberosa, 8 do sexo masculino e 7 do sexo feminino, com idades entre 19 meses e 23 anos; 11 foram considerados como casos esporadicos e, em 4, nao foi possivel precisar os antecedentes familiares.Foram observadas as seguintes manifestacoes cutaneas: adenoma de Pringle em 9 casos; manchas acromicas em 9 casos; fibroma periungueal em 3 casos. Dos 15 casos 13 apresentaram D.M., deficiencia mental, enquanto os outros 2 tinham inteligencia limitrofe. Com excecao de 2 pacientes, todos tinham crises epilepticas e os tipos de crises mais frequentemente observados foram as ausencias atipicas e as crises tonicas. Espasmos infantis estavam presentes em 3 casos e em apenas um caso o eletrencefalograma apresentou aspecto hipsarritmico. A tomografia axial computadorizada, realizada em 7 casos, revelou calcificacoes periventriculares em todos, mesmo naqueles que apresentaram RX simples de cranio normal (4 casos)