Effect of supplementation with algae ß-glucans on performance, health, and blood metabolites of Holstein dairy calves.

Studies have shown that ß-glucans extracted from the cell wall of cereals, algae, and yeasts have been associated with improved immune function. However, it is unknown whether algae ß-glucan supplementation affects the performance, blood metabolites, or cell counts of immune cells in dairy calves. The objective of this randomized clinical trial was to evaluate whether supplementation of ß-glucans to milk replacer in dairy calves fed 6 L/d improved growth performance and fecal status and altered the blood metabolite profile. In this trial, we enrolled Holstein calves (n = 34) at birth (body weight 36.38 ± 1.33 kg; mean ± standard deviation) to receive, from 1 d of age, either 2 g/d algae ß-glucans mixed into 6 L/d of milk replacer (22.4% crude protein and 16.2% fat) or an unsupplemented milk replacer (control). The calves were blocked in pairs according to birth weight, sex, and date of birth (up to 5 d difference). Calves were housed individually, and calf starter (24.7% crude protein and 13.9% neutral detergent fiber) was offered ad libitum based on orts of the previous day until 56 d of age (end of the trial). Body weight was measured weekly, and health checks and daily fecal consistency were evaluated daily in every calf by the same observer. Calves with 2 consecutive days of loose feces that sifted through bedding were considered diarrhea positive. We used a linear mixed effects model to evaluate the effects of ß-glucan supplementation fed during the preweaning period on performance (average daily gain), final weight, feed efficiency (FE), white blood cell count, and selected blood metabolites, repeated by time. A generalized linear mixed effects model was also run to evaluate the likelihood of a diarrhea bout in the first 28 d of life, controlling for the calf as the subject with a logistic distribution. We included age, serum total protein at 48 h, and birth weight as covariates. At 56 d, ß-glucan-supplemented calves weighed more than control calves (56.3 vs. 51.5 kg). Treatment had no effect on total starter intake, but there was a treatment by age interaction for FE, with greater FE for ß-glucan-supplemented calves in wk 3 and 5 of age. There was only a tendency for average daily gain to be greater in supplemented calves than in control calves for the duration of the study. Furthermore, control calves had 14.66 [95% confidence interval (95% CI): 9.87-21.77] times greater odds of having a diarrheal bout than ß-glucan-supplemented calves. Control calves had 12.70 (95% CI: 8.82-18.28) times greater odds of having an additional day with an abnormal fecal score compared with ß-glucan-supplemented calves, suggesting that supplementation ameliorated diarrhea severity. We found no association of treatment with concentrations of serum total protein, albumin, creatinine, or glucose during the preweaning period. Our findings suggest that dietary supplementation of 2 g/d of algae ß-glucans to milk replacer improved fecal status and may affect growth, as evidenced by a higher weaning weight, compared with control calves. Future studies should explore the effect of algae ß-glucans on lower-gut physiology and digestibility in dairy calves.

Characterization of an Infectious Bronchitis Virus Isolated from Commercial Layers Suffering from False Layer Syndrome.

Infectious bronchitis virus (IBV) is a gammacoronavirus that primarily induces an upper respiratory disease in chickens, also affecting the urogenital tract and occasionally leading to a condition called false layer syndrome (FLS), where sexually mature hens ovulate normally but are unable to lay eggs. Here, we describe an outbreak of FLS in Arizona from which an IBV variant that is nearly 90% homologous to DMV/1639 using the Spike subunit 1 gene, named AZ/FLS/17, was isolated and used in challenge experiments. Three-day-old specific-pathogen-free chicks were challenged with AZ/FLS/17 or M41 in high and low doses, and the disease outcomes were compared. Overall, no differences in microscopic lesions or viral loads in the reproductive tract were detected between AZ/FLS/17- and M41-infected birds. To minimize the losses linked to FLS in the problematic flocks, an updated live-attenuated IBV vaccine protocol including the use of the Ma5 strain at the hatchery was implemented, resulting in a drastic reduction of false layers in the subsequent flocks. To monitor the circulation of wild-type and vaccine strains in this population, a molecular surveillance study was performed. Samples were collected at 1, 7, 14, and 21 days of age, and from laying hens at 30 and 36 wk. In older birds, the IBV strains detected were more diverse than at 1 and 7 days of age. Nevertheless, live vaccine combinations are still widely used to decrease the losses caused by FLS in commercial egg laying flocks worldwide.

Caracterización de un virus de la bronquitis infecciosa aislado de aves de postura comerciales que padecían el síndrome de la falsa ponedora. El virus de la bronquitis infecciosa (IBV) es un gammacoronavirus que induce principalmente una enfermedad de las vías respiratorias superiores en los pollos, que también afecta el tracto urogenital y ocasionalmente conduce a una condición llamada síndrome de la falsa ponedora (FLS), donde las gallinas sexualmente maduras ovulan normalmente pero no pueden producir huevos. En este estudio se describe un brote del síndrome de la falsa ponedora en Arizona a partir del cual se aisló una variante del virus de bronquitis que es casi 90% similar a la variante DMV/1639 usando el gene S1, la cual se denominó AZ/FLS/17, y se usó en experimentos de desafío. Pollos de tres días de edad libres de patógenos específicos (SPF) fueron desafiados con el virus AZ/FLS/17 o con el virus M41 en dosis altas y bajas, y se compararon los resultados de la enfermedad. En general, no se detectaron diferencias en las lesiones microscópicas o en las cargas virales en el tracto reproductivo entre las aves infectadas con el virus AZ/FLS/17 y el virus M41. Para minimizar las pérdidas relacionadas con el síndrome de la falsa ponedora en las parvadas problemáticas, se implementó un protocolo actualizado con vacuna viva atenuada contra la bronquitis infecciosa que incluía el uso de la cepa Ma5 en la incubadora, lo que resultó en una reducción drástica de las gallinas falsas ponedoras en las parvadas subsecuentes. Para monitorear la circulación de cepas de tipo silvestre y vacunales en esta población, se realizó un estudio de vigilancia molecular. Se recolectaron muestras a los 1, 7, 14 y 21 días de edad y de gallinas de postura a las 30 y 36 semanas. En aves mayores, las cepas del virus de bronquitis detectadas fueron más diversas que a los días 1 y 7 de edad. Sin embargo, las combinaciones de vacunas vivas todavía se utilizan ampliamente para disminuir las pérdidas causadas por el síndrome de la falsa ponedora en las parvadas de postura de huevo comercial en todo el mundo.

Supplementation of lysolecithin in milk replacer for Holstein dairy calves: Effects on growth performance, health, and metabolites.

Lysolecithin is an antiinflammatory emulsifier associated with improved apparent digestibility of total dietary fat and improved feed efficiency in dairy cattle. However, it is unknown if lysolecithin (LYSO) improves performance in calves. Moreover, since many conventional milk replacers use vegetable-sourced fat (e.g., palm oil), nutrient absorption and fecal score may be affected in neonatal calves. Thus, the objective of this study was to evaluate the effects of LYSO supplemented in milk replacer on performance, metabolites, and gut health of preweaned dairy calves. Holstein calves (n = 32) with adequate passive transfer were assigned in pairs (16 blocks) balanced by birth weight, date of birth, and sex at 1 d of age to randomly receive either LYSO (mixed in 2 milk replacer feedings at a rate of 4 g/d Lysoforte, Kemin Industries Inc., Des Moines, IA) or a milk replacer control (nothing added). Both treatments were fed 6 L/d milk replacer [22.5% crude protein, 16.2% crude fat (vegetable oil fat source) on a dry matter basis with 14% solids] by bucket in 2 daily feedings for 56 d. Calves were individually housed in wooden hutches and offered a commercial calf starter (24.6% crude protein and 13.9% neutral detergent fiber) and water by bucket ad libitum. Feed refusals and calf health was assessed daily. Weights and blood metabolites (glucose, total serum protein, albumin, creatinine, triglycerides, and cholesterol) were sampled weekly, and calves completed the study before weaning at 56 d of age. The effects of LYSO on calf average daily gain, feed efficiency, and blood metabolites were evaluated using a linear mixed model with time as a repeated measure, calf as the subject, and block as a random effect in SAS (SAS Institute Inc., Cary, NC). The effect of LYSO to improve the odds of abnormal fecal score was evaluated using a logistic model. Supplementation of LYSO increased average daily gain (control 0.28 ± 0.03 kg; LYSO 0.37 ± 0.03 kg; least squares means ± standard error of the mean) and increased feed efficiency (gain-to-feed; control 0.25 ± 0.03; LYSO 0.32 ± 0.03). Similarly, LYSO calves had a higher final body weight at d 56 (control 52.11 ± 2.33 kg; LYSO 56.73 ± 2.33 kg). Interestingly, total dry matter intake was not associated with LYSO despite improved average daily gain (total dry matter intake control 1,088.7 ± 27.62 g; total dry matter intake LYSO 1,124.8 ± 27.62 g). Blood glucose, albumin, creatinine, triglycerides, and cholesterol were not associated with LYSO. Indeed, only total serum protein had a significant interaction with LYSO and age at wk 5 and 6. Moreover, control calves had a 13.57 (95% confidence interval: 9.25-19.90) times greater odds of having an abnormal fecal score on any given day during the diarrhea risk period from d 1 to 28. The inclusion of LYSO as an additive in milk replacer in a dose of 4 g/d may improve performance, and calf fecal score, preweaning. Further research should investigate the mechanisms behind the effects of LYSO on fat digestibility in calves fed 6 L/d of milk replacer with vegetable-sourced fat.

Avian Reoviruses of the Same Genotype Induce Different Pathology in Chickens.

The emergence of avian reovirus variant strains has caused negative effects in the poultry industry worldwide. Regardless of the efforts in molecular characterization and classification of these variants, information about the pathogenicity, transmissibility, and immunosuppression in chickens is limited. The genomes of two variant strains (A and B) and a classic S1133 strain (C) belonging to the same sigma C genotype 1 were compared. Additionally, these strains were used in a challenge experiment to evaluate inoculated and indirectly exposed specific-pathogen-free chickens. The whole-genome sequence analysis of the three strains revealed nucleotide identity differences in the L3, M2, and S1 genes. Strains A and B also showed homology differences in the S4 gene, despite having high homologies in all other genes. The in vivo challenge experiments showed that, whereas variant A induced high viral loads in tendons, hearts, and duodena of inoculated chickens, variant B induced high viral loads in indirectly exposed chickens. Likewise, histopathology reflected differences in the pathologic effects induced by these strains. For instance, the B and C strains induced more severe microscopic lesions compared with the A strain. Lymphoid depletion was more severe in bursas than in thymi, and inoculated birds were more affected than exposed birds. In conclusion, different pathologic outcomes in chickens were observed depending on the strain and transmission route. This study provides insights onto the relationship between pathogenicity and genomic composition of avian reoviruses.

Reovirus aviares del mismo genotipo inducen diferentes patologías en pollos. La aparición de cepas variantes del reovirus aviar ha causado efectos negativos en la industria avícola en todo el mundo. Independientemente de los esfuerzos en la caracterización molecular y clasificación de estas variantes, la información sobre la patogenicidad, transmisibilidad e inmunodepresión en pollos es limitada. Se compararon los genomas de dos cepas variantes (A y B) y una cepa S1133 clásica (C) perteneciente al mismo genotipo 1 del gene sigma C. Además, estas cepas se utilizaron en un experimento de desafío para evaluar pollos libres de patógenos específicos inoculados y expuestos indirectamente. El análisis de la secuencia del genoma completo de las tres cepas reveló diferencias de identidad de nucleótidos en los genes L3, M2 y S1. Las cepas A y B también mostraron diferencias de homología en el gene S4, a pesar de tener altas similitudes en todos los demás genes. Los experimentos de exposición in vivo mostraron que, mientras que la variante A inducía altas cargas virales en tendones, corazones y duodeno en los pollos inoculados, la variante B inducía altas cargas virales en pollos expuestos indirectamente. Asimismo, la histopatología reflejó diferencias en los efectos patológicos inducidos por estas cepas. Por ejemplo, las cepas B y C indujeron lesiones microscópicas más graves en comparación con la cepa A. La despoblación linfoide fue más severa en las bolsas que en el timo, y las aves inoculadas resultaron más afectadas que las expuestas. En conclusión, se observaron diferentes resultados patológicos en pollos según la cepa y la vía de transmisión. Este estudio proporciona información sobre la relación entre la patogenicidad y la composición genómica de los reovirus aviares.

Infectious Bronchitis Virus Surveillance in Broilers in California (2012-20).

Infectious bronchitis virus (IBV) causes severe economic losses among chicken flocks worldwide. Although IBV molecular surveillance has been conducted in California broilers, seasonal and spatial-temporal trends in IBV prevalence are poorly defined. The goals of this study were to evaluate seasonal and spatial-temporal trends in IBV prevalence and to determine the predominant IBV genotypes obtained over the last 8 yr from a broiler company located in the California Central Valley. In total, 3439 broilers with a suspicion of IBV infection were submitted to the California Animal Health and Food Safety laboratories between January 2012 and February 2020. Swabs from tracheas, kidneys, and cecal tonsils from each submission were independently pooled and screened for IBV using reverse transcriptase quantitative PCR (RT-qPCR). Positive samples were submitted for virus isolation. Viral isolates were subject to a conventional RT-PCR targeting the S1 gene hypervariable region. Positive samples from this RT-PCR were sequenced, and phylogenetic analyses were performed. In total, 1243 pooled swab samples were positive for IBV. Positive results were more frequently detected in fall and winter months compared to spring. Spatial analyses revealed an IBV hot spot in the vicinity of Livingston, and two areas with a low prevalence (i.e., cold spots) around Riverdale. The IBV spatial-temporal distribution identified three significant clusters: one hot spot around Turlock from 2015 to 2016, a second hot spot around Merced from 2012 to 2016, and a cold spot around Fresno from 2017 to 2020. Predominant genotypes changed over time from IBV Cal 99, which was predominant between 2012 and 2014, to IBV 3099 in 2019. Vaccination efforts were initiated in 2018, and as a result, we detected an emerging variant with 92% similarity to CA 3099 in 2020. This work highlights the importance of ongoing surveillance in IBV prevention programs. Surveillance strategies are necessary to monitor trends in diseases such as infectious bronchitis, and the tools used for surveillance need to be sensitive enough to detect new variants and identify spatial-temporal trends.

Vigilancia del virus de la bronquitis infecciosa en pollos de engorde en California (2012­2020). El virus de la bronquitis infecciosa (IBV) causa graves pérdidas económicas entre las parvadas de pollos en todo el mundo. Aunque la vigilancia molecular del virus de la bronquitis infecciosa se ha realizado en pollos de engorde en el estado de California, las tendencias estacionales y espacio-temporales sobre la prevalencia de este virus están mal definidas. Los objetivos de este estudio fueron evaluar las tendencias estacionales y espacio-temporales sobre la prevalencia del virus de la bronquitis infecciosa y determinar los genotipos predominantes de este virus obtenidos durante los últimos ocho años de una empresa de pollos de engorde ubicada en el Valle Central de California. En total, 3439 pollos de engorde con sospecha de infección por el virus de la bronquitis infecciosa se enviaron a los laboratorios de Salud Animal y Seguridad Alimentaria del estado de California entre enero del 2012 y febrero del 2020. Los hisopos de tráqueas, riñones y tonsilas cecales de cada caso se combinaron de forma independiente y se examinaron para detectar al virus de la bronquitis utilizando transcripción reversa y un método cuantitativo de PCR (RT-qPCR). Se enviaron muestras positivas para aislamiento del virus. Los aislados virales se sometieron a un método convencional de RT-PCR dirigido a la región hipervariable del gene S1. Se secuenciaron muestras positivas mediante la prueba RT-PCR y se realizaron análisis filogenéticos. Un total de 1243 muestras combinadas de hisopos dieron positivo para el virus de la bronquitis infecciosa. Los resultados positivos se detectaron con mayor frecuencia en los meses de otoño e invierno en comparación con la primavera. Los análisis espaciales revelaron un punto activo para el virus de la bronquitis infecciosa en las cercanías de Livingston y dos áreas con una baja prevalencia (es decir, puntos fríos) alrededor de Riverdale. La distribución espacio-temporal del virus de la bronquitis identificó tres grupos importantes: un punto activo alrededor de Turlock entre los años 2015 a 2016, un segundo punto activo alrededor de Merced entre los años 2012 a 2016 y un punto frío alrededor de Fresno entre los años 2017 a 2020. Los genotipos predominantes cambiaron con el tiempo, consideraron el subtipo IBV Cal 99, que fue predominante entre 2012 y 2014, a el tipo IBV 3099 en 2019. Los esfuerzos de vacunación se iniciaron en el 2018, y como resultado, detectamos una variante emergente con un 92% de similitud con el virus CA 3099 en 2020. Este trabajo destaca la importancia de la vigilancia continua en los programas de prevención para la bronquitis infecciosa. Las estrategias de vigilancia son necesarias para monitorear las tendencias en enfermedades como la bronquitis infecciosa, y las herramientas utilizadas para la vigilancia deben ser lo suficientemente sensibles como para detectar nuevas variantes e identificar tendencias espacio-temporales.

Whole-flint corn grain or tropical grass hay free choice in the diet of dairy calves.

Tropical grass hay feeding is related to improved ruminal health; however, it may decrease energy intake. On the other hand, whole-flint corn grain may be an alternative fiber source in the diet of dairy calves. Forty-two Holstein calves were used in a randomized block design, considering sex, birth date, and weight at 21 d of age, when the supply of whole-flint corn grain or tropical grass hay started. Three component-fed solid diets were compared: (1) starter concentrate only during the preweaning and starter concentrate with free choice of chopped Tifton-85 hay postweaning (SC), (2) starter concentrate with free choice of chopped Tifton-85 hay pre- and postweaning (SCH), and (3) starter concentrate with free choice of whole-flint corn grain pre- and postweaning (SCW). The animals were evaluated from 21 to 84 d of age. Calves were managed equally during the first 21 d, fed with 6 L/d of whole milk and a commercial starter concentrate (46% nonfiber carbohydrates, small particles, and pelleted) ad libitum. After that, milk feeding was reduced to 4 L/d until gradual weaning at 56 d of age. At 56 d of age, 4 animals per treatment were randomly chosen to be slaughtered for digestive tract weight evaluation and to collect tissue for histological analysis of the ruminal wall, duodenum, and cecum, whereas the other 30 animals were weaned and evaluated for a further 22 d when the SC diet also received hay ad libitum. Feed intake was measured daily. Weight gain and metabolic indicators of intermediate metabolism were evaluated weekly. Ruminal fluid was collected at wk 6, 8, 10, and 12 of age. The SCH diet increased the total and starter dry matter intake, and consequently, the average daily gain and body weight at 56 d of age. The SCW diet promoted an increase in propionate and decreased acetate-to-propionate ratio. Morphometric variables were affected by the SCH diet. The postweaning performance was unaffected by solid diets; however, the SCW diet decreased ruminal and fecal pH. Feeding hay, starting at 21 d of age, can stimulate early solid diet intake, promoting better performance and ruminal and intestinal development, when a highly fermentable and small particle pelleted starter is fed.

Infectious Coryza: Persistence, Genotyping, and Vaccine Testing.

The reemergence of infectious coryza (IC) caused by Avibacterium paragallinarum (AP) as an acute and occasionally chronic respiratory disease in domestic poultry has caused severe losses in several U.S. states. The disease is also associated with decreased egg production in layers and increased condemnations from air sac infections in broilers. A series of applied experiments were performed to elucidate the persistence of AP in infected broiler flocks, to genotype AP strains isolated from field cases, and to evaluate commercial and autogenous vaccine protection in commercial and specific-pathogen-free (SPF) chickens. Experimental evaluation of environmental persistence suggests that AP did not persist more than 12 hr in a hypothetically contaminated environment. Additionally, other detected potential pathogens such as Gallibacterium anatis and infectious bronchitis virus caused mild respiratory signs in the exposed birds. The HMTp210 and HagA genes of four IC field strains were sequenced and compared with published sequences of HMTp210 and HagA. The HMTp210 phylogeny showed a marginally imperfect clustering of the sequences in genogroups A, B, and C. Although not definitive, this phylogeny provided evidence that the four field strains aligned with previously characterized serovar C strains. Moreover, the base pair homology of the four strains was 100% identical to serovar C reference strains (H-18 and Modesto). HagA phylogeny was unclear, but interestingly, the IC field strains were 100% homologous to C-1 strains reported from Mexico and Ecuador. Finally, vaccine protection studies in commercial hens indicate that clinical signs are induced by a combination of IC and other concomitant pathogens infecting commercial birds. Additionally, vaccine protection experiments performed in SPF hens indicated that protection provided by the two commercial vaccines tested provided a reduction in clinical signs and bacterial shedding after two applications.

Coriza infecciosa: Persistencia, genotipificación y pruebas para vacunas. El resurgimiento de la coriza infecciosa (CI) causada por Avibacterium paragallinarum (AP) como una enfermedad respiratoria aguda y ocasionalmente crónica en aves domésticas ha causado graves pérdidas en varios estados de los Estados Unidos. La enfermedad también se asocia con una disminución en la producción de huevo en gallinas de postura y al incremento de decomisos por infecciones de los sacos aéreos en pollos de engorde. Se realizó una serie de experimentos aplicados para dilucidar la persistencia de A. paragillanarum en parvadas de pollos de engorde infectados, para genotipificar las cepas de A. paragallinarum aisladas de casos de campo y para evaluar la protección de vacunas comerciales y autógenas en pollos comerciales y en aves libres de patógenos específicos (SPF). La evaluación experimental de la persistencia ambiental sugiere que A. paragallinarum no persistió más de doce horas en un ambiente hipotéticamente contaminado. Además, otros patógenos potenciales detectados como Gallibacterium anatis y el virus de la bronquitis infecciosa causaron signos respiratorios leves en las aves expuestas. Los genes HMTp210 y HagA de cuatro cepas de campo de coriza infecciosa se secuenciaron y compararon con las secuencias publicadas de HMTp210 y HagA. La filogenia de HMTp210 mostró una agrupación marginalmente imperfecta de las secuencias en los genogrupos A, B y C. Aunque no es definitiva, esta filogenia proporcionó evidencia de que las cuatro cepas de campo se alinearon con cepas del serovar C previamente caracterizadas. Además, la homología de pares de bases de las cuatro cepas fue 100% idéntica a las cepas de referencia del serovar C (H-18 y Modesto). La filogenia de HagA no fue clara, pero curiosamente, las cepas de campo de coriza infecciosa fueron 100% similares con las cepas C-1 reportadas en México y Ecuador. Finalmente, los estudios de protección de vacunas en gallinas comerciales indican que los signos clínicos son inducidos por una combinación de coriza infecciosa y otros patógenos concomitantes que infectan a las aves comerciales. Además, los experimentos de protección de vacunas realizados en aves libres de patógenos específicos indicaron que la protección proporcionada por las dos vacunas comerciales analizadas proporcionó una reducción en los signos clínicos y en la eliminación bacteriana después de dos aplicaciones.

Understanding Immune Resistance to Infectious Bronchitis Using Major Histocompatibility Complex Chicken Lines.

Genetic resistance or susceptibility to infectious diseases has been largely associated with the avian major histocompatibility complex (MHC) genes. Our goal was to determine resistance and susceptibility of MHC B haplotype in congenic and inbred chicken lines in order to establish a resistant-susceptible model. Eight congenic lines (253/B18, 254/B15, 330/B21, 312/B24, 331/B2, 335/B19, 336/B21, and 342/BO), two inbred lines (003/B17 and 077/B19), and three commercial lines (white leghorn, brown layers, and broilers) were used in two experiments. We analyzed and compared immunologic responses and the effect of challenge by measuring viral load, IgG and IgA humoral responses, histopathology and histomorphometry, clinical signs, and immune cell populations in the different MHC B haplotype lines. We found that respiratory signs, tracheal deciliation and inflammation, airsacculitis, viral shedding in tears, and local humoral responses were good parameters to determine resistance or susceptibility. Based on these results, we identified 331/B2 as the most resistant and 335/B19 as the most susceptible congenic chicken lines. These two lines will be used as an animal model in subsequent experiments to understand the mechanisms by which the immune system in chickens generates resistance to infectious bronchitis virus.

Sarcopenia and osteoporosis in Portuguese centenarians.

BACKGROUND/OBJECTIVES: To evaluate the prevalence of sarcopenia and osteoporosis in centenarians and its association with body composition and past physical activity (PPA). SUBJECTS/METHODS: We conducted an observational cross-sectional study in a representative nationwide population of 253 Portuguese centenarians. A clinically oriented questionnaire and anthropometric and body composition analysis by bioimpedance were executed. Sarcopenia was determined using a muscle mass (MM) index cutoff ⩽16.7 kg/m2. Osteoporosis was defined through estimated bone mass (BM), according to gender and body weight. Inadequate physical performance was considered when the timed-up-and-go test was >12 s. RESULTS: BM and MM were gender dependent, even after adjusting for multiple variables. We found a protective effect of intense PPA on sarcopenia risk (OR=0.41, 95% CI=0.20-0.82, P=0.013). Those with worse physical function status presented a significantly higher risk for osteoporosis (OR=8.9, 95% CI=1.94-40.08, P=0.005). Multivariate models showed that low MM, BM and fat mass (FM) were predictors of underweight in centenarians. Underweight, male gender, worst physical function, hypohydration and sarcopenia were associated with increased osteoporosis risk. The risk for sarcopenia was higher in women (P=0.002), in those underweight (P=0.005) and in osteoporotic individuals (P<0.0001). Conversely, increased MM was protective against osteoporosis (P<0.0001), whereas higher BM decreased the risk for sarcopenia (P=0.005). CONCLUSIONS: Gender was revealed to be a relevant factor influencing BM and MM in centenarians. MM and BM were better predictors of underweight than was dehydration. Healthy BM and MM were positively related with PPA intensity and determined physical performance, having a positive impact on the centenarian's independence. During aging, improving physical function might add to sarcopenia and osteoporosis prevention, besides prevention of femur neck fractures and cardiovascular diseases.

Evaluation of Recurrence Predictors and Survival Probability After Liver Transplantation for Hepatocellular Carcinoma: Analysis From a Single Center.

BACKGROUND: Liver transplantation is the criterion standard treatment for hepatocellular carcinoma, but tumor recurrence remains a problem leading to a worse prognosis. We aimed to evaluate factors associated with recurrence of hepatocellular carcinoma and survival after this procedure. METHODS: We retrospectively reviewed data from 101 patients submitted to liver transplantation because of hepatocellular carcinoma from January 2005 to January 2012 at our single center. Univariate and multivariate analyses were performed to identify preoperative factors and histologic findings associated with lower survival rates and recurrence. The significance level was 5%. RESULTS: There was recurrence in 10 cases (9.9%), with an average time of 25.28 ± 26.92 months. Microvascular invasion (P = .005; hazard ratio [HR], 4.94; 95% confidence interval [CI], 1.42-17.12) was an independent factor for recurrence. Microvascular invasion (P = .035; HR, 1.87; 95% CI, 1.04-3.25) and tumors outside the criteria of the University of San Francisco (P = .046; HR, 1.81; 95% CI, 1.01-3.25) were independent factors for the risk of death. Poorly differentiated tumors had a higher level of alphafetoprotein (P = .03), and values <100 ng/mL were associated with well differentiated tumors. CONCLUSIONS: Microvascular invasion was associated with recurrence and lower survival. Tumors outside the criteria of the University of California, San Francisco had lower survival. Alpha-fetoprotein levels >100 ng/mL were associated with poorly differentiated tumors.

Characterization and genetic diversity of pepper (Capsicum spp) parents and interspecific hybrids.

Pepper species exhibit broad genetic diversity, which enables their use in breeding programs. The objective of this study was to characterize the diversity between the parents of different species and their interspecific hybrids using morphological and molecular markers. The parents of Capsicum annuum (UFPB-01 and -137), C. baccatum (UFPB-72), and C. chinense (UFPB-128) and their interspecific hybrids (01x128, 72x128, and 137x128) were used for morphological and molecular characterization. Fruit length and seed yield per fruit (SYF) traits showed the highest variability, and three groups were formed based on these data. CVg/CVe ratio values (>1.0) were calculated for leaf length (1.67) and SYF (5.34). The trait that most contributed to divergence was the largest fruit diameter (26.42%), and the trait that least contributed was pericarp thickness (0.33%), which was subject to being discarded. The 17 primers produced 58 polymorphic bands that enabled the estimation of genetic diversity between parents and hybrids, and these results confirmed the results of the morphological data analyses. The principal component analysis results also corroborated the morphological and random-amplified polymorphic DNA data, and three groups that contained the same individuals were identified. These results confirmed reports in the literature regarding the phylogenetic relationships of the species used as parents, which demonstrated that C. annuum was closer to C. chinense as compared to C. baccatum.

Evaluation of a histocompatibility antigen related to hepatitis B virus in patients with hepatocellular carcinoma in the western Brazilian Amazon.

Hepatocellular carcinoma is an infection of variable incidence that can be caused by hepatitis B virus (HBV), which is endemic in the Amazon region. The diagnosis of HBV can be performed through the use of serum markers such as the hepatitis B surface antigen. The chronic HBV can cause mutagenesis and carcinogenesis, being the susceptibility of infection due to allele human leukocyte antigen (HLA). Thus, we evaluated the clinical, molecular and laboratory profile (histocompatibility complex) of HBV in 22 patients with hepatocellular carcinoma in Amazonia, including 18 males and 4 females, using a blood sample for generic HLA class II. The results showed increased frequency of disease evolution in adults between 25 and 64 years old, who comprised 19 of the 22 patients studied. Most patients (16/22) presented high levels of alpha-fetoprotein and transaminases (14/22). The most common HLA alleles were DRB1 04 (8/44), DRB1 08 (9/44), DRB 03 (16/44), and DQB1 04 (9/44). When we compared specific phenotype frequencies of HLA-DRB1 between patients and controls, we found that patients had a significantly higher frequency of allele DRB1 08 and a significantly lower frequency of DRB1 07 and DRB1 12 compared to previous studies on Asian and Amazonian populations suggesting ethnic differences. We suggest that alleles HLA-DRB 08, HLA-DRB 03 and HLA-DQB1 04 may be risk factors for hepatocellular carcinoma in Amazon.

Motor behavioral abnormalities and histopathological findings of Wistar rats inoculated with HTLV-1-infected MT2 cells.

The objective of the present study was to describe motor behavioral changes in association with histopathological and hematological findings in Wistar rats inoculated intravenously with human T-cell lymphotropic virus type 1 (HTLV-1)-infected MT2 cells. Twenty-five 4-month-old male rats were inoculated with HTLV-1-infected MT2 cells and 13 control rats were inoculated with normal human lymphocytes. The behavior of the rats was observed before and 5, 10, 15, and 20 months after inoculation during a 30-min/rat testing time for 5 consecutive days. During each of 4 periods, a subset of rats was randomly chosen to be sacrificed in order to harvest the spinal cord for histopathological analysis and to obtain blood for serological and molecular studies. Behavioral analyses of the HTLV-1-inoculated rats showed a significant decrease of climbing, walking and freezing, and an increase of scratching, sniffing, biting, licking, and resting/sleeping. Two of the 25 HTLV-1-inoculated rats (8%) developed spastic paraparesis as a major behavioral change. The histopathological changes were few and mild, but in some cases there was diffuse lymphocyte infiltration. The minor and major behavioral changes occurred after 10-20 months of evolution. The long-term observation of Wistar rats inoculated with HTLV-1-infected MT2 cells showed major (spastic paraparesis) and minor motor abnormalities in association with the degree of HTLV-1-induced myelopathy.

Observation of tags and hybrid layer of a single bottle conventional adhesive system and a self-etching adhesive system, on sound dentin.

This experimental light microscopy study investigated the formation of a hybrid layer and resin tags on sound dentin, after utilization of conventional and self-etching adhesive systems. After restorative procedures, the specimens were decalcified in a formic acid and sodium citrate solution, embedded in paraffin, sectioned at 6-microm thickness and stained by the Brown & Brenn method for analysis and measurement by light microscopy (AXIOPHOT) (400x). The results were statistically analyzed by analysis of variance, at a significance level of 5%. Based on the results, it could be concluded that the conventional adhesive allowed the formation of a thicker hybrid layer than the self-etching adhesive, with similar penetration into the dentinal tubules (resin tags).

Low molecular weight protein tyrosine phosphatase genetic polymorphism and susceptibility to cancer development.

Low molecular weight protein tyrosine phosphatases (LMW-PTPs) are a family of 18-kDa enzymes involved in cell growth regulation. Human acid phosphatase 1 (ACP1) is genetically polymorphic, and three common alleles segregating at the ACP1 locus on the short arm of chromosome 2 give rise to six phenotypes. Each allele appears to encode two electrophoretically different isozymes, fast and slow, which are produced in allele-specific ratios. Fast isozymes are related with cytoskeletal organization, cellular organization, and spreading. Slow isozymes are associated with growth factor receptors and dephosphorylation. In this study, ACP1 genetic polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism on 74 subjects with various cancers; the control group was 236 healthy subjects randomly selected. With genotypes cumulated according to fast isoform concentration, [A + AC] < [AB + BC] < [BB], subjects with cancer presented an increase of fast isozyme concentration (BB 38.2%; P = 0.002, chi2), relative to the control sample (19.8%). The increase of fast isozyme concentration increased the invasive capacity of cancer cells, whereas a decrease of slow isozyme concentration in cancer did not cause growth inhibition and so resulted in cancer cell proliferation.

Immunization with cytoplasmic repetitive antigen and flagellar repetitive antigen of Trypanosoma cruzi stimulates a cellular immune response in mice.

In previous studies, we demonstrated that CRA and FRA recombinant proteins, used for diagnosis of Chagas' disease, elicited a humoral immune response in susceptible and resistant mice. To understand better the immune response to these proteins, we have evaluated, the cellular immune response in CRA- and in FRA-immunized BALB/c and C57BL/6 mice. A specific cellular lymphoproliferative response was observed in both strains of mice. Spleen cell cultures mainly from CRA-immunized C57BL/6 and FRA-immunized BALB/c mice produced high levels of IFN-y, indicating the induction of a Type 1 immune response. Regarding the T cell subsets, CD4+ T cells were the major source of IFN-y in CRA- and FRA-immunized mice. These results suggest that CRA and FRA are important immunogens in inducing a Type 1 immune response and that they may be considered as potential vaccine antigens.

Interleukin-12 stimulation of lymphoproliferative responses in Trypanosoma cruzi infection.

The cytokine interleukin-12 (IL-12) is essential for resistance to Trypanosoma cruzi infection because it stimulates the synthesis of interferon-gamma (IFN-gamma), a major activator of the parasiticidal effect of macrophages. A less studied property of IL-12 is its ability to amplify the proliferation of T or natural killer (NK) lymphocytes. We investigated the role of endogenously produced IL-12 in the maintenance of parasite antigen (T-Ag)-specific lymphoproliferative responses during the acute phase of T. cruzi infection. We also studied whether treatment with recombinant IL-12 (rIL-12) would stimulate T-Ag-specific or concanavalin A (Con A)-stimulated lymphoproliferation and abrogate the suppression that is characteristic of the acute phase of infection. Production of IL-12 by spleen-cell cultures during T. cruzi infection increased in the first days of infection (days 3-5) and decreased as infection progressed beyond day 7. The growth-promoting activity of endogenous IL-12 on T-Ag-specific proliferation was observed on day 5 of infection. Treatment of cultures with rIL-12 promoted a significant increase in Con A-stimulated proliferation by spleen cells from normal or infected mice. Enhanced T-Ag-specific proliferation by rIL-12 was seen in spleen cell cultures from infected mice providing that nitric oxide production was inhibited by treatment with the competitive inhibitor NG-monomethyl-L-arginine (NMMA). Enhancement of proliferation promoted by IL-12 occurred in the presence of neutralizing anti-interleukin-2 (IL-2) antibody, suggesting that this activity of IL-12 was partly independent of endogenous IL-2. Thymidine incorporation levels achieved with rIL-12 treatment of the cultures were approximately 50% of those stimulated by rIL-2 in the same cultures.

The influence of differing pore sizes on the biocompatibility of two polypropylene meshes in the repair of abdominal defects. Experimental study in dogs.

Two types of monofilament polypropylene meshes of markedly different construction, configuration and pore size were compared and used to repair full-thickness muscle defects in the abdominal wall of 22 mongrel dogs to assess their biocompatibility with host tissues. The defects were repaired with Prolene (Ethicon) woven mesh (pore size = 164 x 96 microns) and with an experimental, extruded mesh called T mesh (pore size = 3 mm x 4 mm). On the 30th postoperative day, the animals were sacrificed, and the segments of the abdominal wall containing the implanted meshes were excised. Although the Prolene mesh had greater tensile strength before implantation, 30 days after implantation, the T mesh showed similar tensile strength to Prolene mesh. The collagen densitometry showed a significant increase of total and mature collagen type I deposition in the T mesh. This suggests that the increased mature collagen type I deposition significantly increases the tensile strength of the reinforced mesh tissue and that the larger pore in the T mesh contributed to this finding by allowing increasing fibber orientation within the pores as a result of in vivo tension.