Effect of low-protein corn and soybean meal-based diets on nitrogen utilization, litter quality, and water consumption in broiler chicken production: insight from meta-analysis.

The growing demand for high-value animal protein must be met using sustainable means that optimize the utilization of nutrients, especially nitrogen (N) so that excreta do not over-fertilize fields and end up causing soil acidification, waterway eutrophication and greenhouse gas emissions. Malodorous N compounds can cause respiratory diseases and poor growth in livestock. The increasing availability of feed-grade amino acids makes it possible to formulate low-protein diets for broilers and thereby reduce N excretion. However, published studies of the effects of such diets on broiler growth performance have been based on reducing CP contents gradually in a variety of ways that have given inconsistent results. Since the amount of published data is now large, a meta-analysis was performed in order to categorize diet formulation strategies and quantify their impact on N balance, water consumption, litter moisture, plasma uric acid. This showed that lowering the CP content of broiler diets generally means replacing some soybean meal with corn and hence increasing the starch content. However, since soybean meal is also a source of potassium, this reduces electrolyte balance. Lowering the CP content from 19% to 17% is associated with a 29% reduction of N excretion in broilers aged 0-21 d, and a 7% increase in N efficiency (N retention/N intake). Reducing the CP content from 19% to 17% decreases daily water consumption by 20.6 mL/bird, litter moisture by 2.2% and plasma uric acid by 0.56 mg/dL. This meta-analysis improves our understanding of the low-protein strategy and allows us to quantify its impact on N balance, litter quality and uric acid. It shows that managing N excretion is wholly beneficial and reduces litter wetness.

Effects of Bacillus subtilis, butyrate, mannan-oligosaccharide, and naked oat (ß-glucans) on growth performance, serum parameters, and gut health of broiler chickens.

Four nonantibiotic alternative growth promoters for broiler chickens were evaluated. Ross 308 chicks were fed a control diet (mainly corn and soybean meal) or a diet supplemented with a probiotic (Bacillus subtilis Gallipro DSM 17299), encapsulated butyric acid (Novyrate C), mannan-oligosaccharide (Actigen MOS) or formulated with 20% naked oat (starter diet) and 30% naked oat (grower and finisher). The study was carried out as a complete random blocked design with 10 pens for each diet, 45 birds per pen. Compared to the control, the naked oat diet improved the average daily gain by 16% during the starter phase (up to d 10). The probiotic did so during the grower phase as did butyric acid in the finisher phase (up to d 34). For the experiment overall, the probiotic decreased average daily gain slightly. The best improvement in feed conversion ratio was obtained in the butyrate group (5%). No significant treatment effect on crop pH or on mortality was observed. The naked oat diet gave a slightly lower cecum pH on d 34. The MOS supplement decreased jejunal mass on d 34 and increased villus length (34%) and villus height/crypt depth ratio (32%) measured on d 10. Naked oat, butyric acid and MOS diets all reduced serum endotoxin levels. The probiotic increased serum C-reactive protein. All noncontrol diets reduced serum malondialdehyde. The naked oat diet reduced d 34 litter pH by about 0.3. Some effects of the proposed non-antibiotic growth promoters have been observed and could contribute to livestock performance. Their exact modes of action remained to be defined.

Cervical vertebral osteomyelitis secondary to Streptococcus equi infection in an adult horse - case report / Osteomielite vertebral cervical secundária à infecção por Streptococcus equi em um cavalo adulto - relato de caso

A 15-year-old, mixed breed, male horse was attended with a history of multiple abscesses in the cervical region with a three-year evolution. Upon admission, three fistulous tracts with drainage of purulent secretions in the cervical region, low body score, restriction of cervical movements, and painful sensitivity to palpation were observed. The horse was diagnosed with osteomyelitis secondary to Streptococcus equi infection. The initial treatment was antibiotic therapy and local curative. Owing to the lack of response, surgical debridement was performed. An initial favorable response was observed; however, after 4 months, drainage recurred, and the animal was euthanized. A post-mortem computed tomography scan was performed to obtain details of the injury. Cervical osteomyelitis is rare, and its occurrence through hematogenous spread in adult horses and the tomographic findings had not been reported previously. The long period of evolution, difficulty in performing an aggressive debridement, and the presence of multi-drug resistant bacteria contributed to the negative outcome.(AU)

Um equino macho, sem raça definida, de 15 anos de idade, foi atendido com histórico de múltiplos abscessos cervicais com evolução de três anos. Na admissão, foram observados: três trajetos fistulosos com drenagem de material purulento na região cervical; baixo escore corporal; restrição de movimentos cervicais; e sensibilidade dolorosa à palpação da região. Foi diagnosticada osteomielite vertebral cervical secundária à infecção por Streptococcus equi. O tratamento inicial consistiu na administração de antibióticos e curativo local. Na ausência de resposta à terapia, realizou-se o debridamento cirúrgico. Inicialmente, obteve-se uma resposta favorável, entretanto, após quatro meses, houve recidiva da lesão e o animal foi submetido à eutanásia. Realizou-se tomografia computadorizada no post mortem para detalhamento da lesão. A osteomielite vertebral cervical é rara, e sua ocorrência por meio de disseminação hematógena em animais adultos não foi previamente reportada. O longo período de evolução, aliado à dificuldade de realização de um debridamento agressivo, e a característica multirresistente do agente etiológico contribuíram para o desfecho negativo do caso.(AU)

Interaction of potassium carbonate and soybean oil supplementation on performance of early-lactation dairy cows fed a high-concentrate diet.

Potassium carbonate supplementation is known to improve milk fat synthesis and to modify milk mineral composition in dairy cows. The objective of the current experiment was to evaluate the effect of K2CO3 on production performance, biohydrogenation of fatty acids (FA), and mineral composition of milk in early-lactation dairy cows fed a high-concentrate diet with or without soybean oil (SBO), as a source of polyunsaturated FA. Twenty-eight ruminally fistulated Holstein cows were used in a randomized complete block design. The experiment lasted 33 d, including a 5-d pretreatment collection period used as a covariate. Experimental treatments were arranged as a 2 × 2 factorial with 0 or 1.5% K2CO3 and with 0 or 2% SBO, and balanced to contain 40% forage (57% corn silage + 43% grass silage) and 60% concentrate. Preplanned orthogonal contrasts were used to assess the effects of K2CO3, SBO, and their interaction. Feeding K2CO3 did not affect milk yield, but tended to increase 4% fat-corrected milk and fat yield when combined with SBO. However, adding SBO to diets increased milk yield. Dietary K2CO3 supplementation did not affect milk fat concentration of trans-10 18:1 or any other identified biohydrogenation intermediates. Soybean oil supplementation decreased milk fat concentration of C16 and de novo synthesized FA, and increased preformed FA. Among the other effects of SBO supplementation observed, concentrations of cis-9,trans-11 18:2 increased, as well as most of the cis and trans isomers of 18:1 and 18:0. Milk urea N decreased in cows fed K2CO3 as compared with unsupplemented diets. A positive relation was established between milk Cl concentration and milk yield, suggesting that the equilibrium of this ion is linked to the efficiency of lactogenesis. The effect of K2CO3 on this mineral equilibrium in the mammary gland remains to be established. Overall, results have shown that potential effect of K2CO3 on milk fat synthesis is dependent on the levels of dietary polyunsaturated FA.

Potassium carbonate as a cation source for early-lactation dairy cows fed high-concentrate diets.

Previous studies reported that addition of K2CO3 to high-concentrate diets improved milk fat synthesis, although the mechanism is yet to be established. The objective of the current experiment was to investigate the effects of dietary cation-anion difference (DCAD), cation source, and buffering ability of the mineral supplement on rumen biohydrogenation of fatty acids and production performance in dairy cows fed a high-concentrate diet. Thirty-five early-lactation Holstein cows (25 multiparous ruminally fistulated and 10 primiparous nonfistulated) were used in a randomized complete block design (7 blocks) with 33-d periods, including a 5-d pre-treatment collection period used as a covariate. Diets were (1) control, a basal diet [47% nonfibrous carbohydrates, DCAD (Na + K - Cl - S) = 65 mEq/kg of dry matter (DM)] containing 40% forage (including 60% corn silage) and 60% concentrate, (2) K2CO3 (control + K2CO3, 1.8% of DM, DCAD = 326 mEq/kg of DM), (3) KHCO3 (control + KHCO3, 2.6% of DM, DCAD = 324 mEq/kg of DM), (4) KCl (control + KCl, 2.0% of DM, DCAD = 64 mEq/kg of DM), and (5) Na2CO3 (control + Na2CO3, 1.4% of DM, DCAD = 322 mEq/kg of DM). Pre-planned orthogonal contrasts were used to assess the effects of K2CO3 (control vs. K2CO3), buffering ability (K2CO3 vs. KHCO3), DCAD (K2CO3 vs. KCl), and cation type (K2CO3 vs. Na2CO3). Supplementing K2CO3 in a high-concentrate diet did not improve milk fat yield or 4% fat-corrected milk yield. Milk fat concentration was greater in cows fed K2CO3 compared with control (4.03 vs. 3.26%). Milk yield tended to decrease (34.5 vs. 38.8 kg/d) and lactose yield decreased in cows fed K2CO3 as compared with KCl (1.64 vs. 1.87 kg/d). Milk fat concentration of trans-10 18:1 was increased when cows were fed Na2CO3 as compared with K2CO3. A positive relationship was observed between concentrations of anteiso 15:0 and trans-10,cis-12 18:2 in milk fat from cows receiving K2CO3. Milk Na concentration was increased, whereas milk Cl was decreased with K2CO3 as compared with KHCO3 or KCl. A positive relationship was established between milk Cl concentration and milk yield (R2 = 0.34) across all dietary treatments. Cation-anion difference (Na + K - Cl - S) in ruminal fluid was increased with K2CO3 as compared with control or KCl. Blood pH tended to decrease in cows fed KCl compared with K2CO3. Our results suggest that mineral supplementation tends to affect milk and milk fat synthesis and that factors other than DCAD, potassium ion, or buffer ability may be implicated. The variations observed in mineral composition of milk suggest an allostatic process to maintain an ionic equilibrium in mammary epithelial cells in response to mineral composition of the diet.

Kidney transplantation in children weighing less than 15 kg: extraperitoneal surgical access-experience with 62 cases.

Small children are a challenging group in whom to perform KT. This retrospective study analyzed the results of 62 KTs in children weighing <15 kg, performed between 1998 and 2010, using extraperitoneal access and anastomosis of the renal vessels of donors to the aorta and IVC or iliac vessels of the recipients. Thirty-two (51.6%) grafts were LRDTs and 30 (48.4%) were DDRTs-28 of them pediatric. The mean age at KT was 3.7 ± 2.2 yr (1-12), and the mean weight was 12.3 ± 2.1 kg (5.6-14.9). Ten children weighed <10 kg, and five (8.1%) children presented previous thrombosis of the venous system. At one and five yr, patient survival was 93.2% and 84.2%, and graft survival was 85.2% and 72.7%. There were no differences between the rates for LRDT and DDRT. There were six vascular complications (four vascular thromboses, one laceration, and one renal artery stenosis) and two perirenal collections. Extraperitoneal access is a valid KT technique in children weighing <15 kg.

Proteomic analysis of metacyclic trypomastigotes undergoing Trypanosoma cruzi metacyclogenesis.

Trypanosoma cruzi, the causative agent of the Chagas disease, has a complex life cycle alternating between replicative and noninfective forms with nonreplicative and infective forms of the parasite. Metacyclogenesis is a process that takes place in the invertebrate host, comprising morphogenetic transformation from a noninfective form to an infective form, such that parasites acquire the ability to invade human cells. We analyze here the metacyclogenesis process by 2D electrophoresis coupled to MALDI-TOF MS. A large proportion of unique proteins expressed during metacyclogenesis were observed. Interestingly, 50% of the spots were found to differ between epimastigotes and trypomastigotes. We provide a 2D map of the infective metacyclic trypomastigotes. Sixty six protein spots were successfully identified corresponding to 43 different proteins. We analyzed the expression profiles for the identified proteins along metacyclogenesis and classified them into three groups according to their maximal level of expression. We detected several isoforms for a number of proteins, some displaying differential expression during metacyclogenesis. These results suggest that posttranslational modifications may be a fundamental part of the parasite's strategy for regulating gene expression during differentiation. This study contributes to the identification of relevant proteins involved in the metacyclogenesis process. The identification and molecular characterization of these proteins will render vital information about the steps of the parasite differentiation into the infective form.

Results of en bloc renal transplants of pediatric deceased donors into adult recipients.

UNLABELLED: Various strategies have evolved to expand the donor pool due to the extreme shortage of organs. Herein we reviewed our experience with en bloc pediatric kidney transplantation since 1998. METHODS: From January 1998 to December 2004, nine adult patients underwent kidney transplantation using en bloc kidneys from donors <5 years old (range, 1 to 4). The mean age of the recipients was 45.1 years (range, 34 to 57). RESULTS: In recipients of en bloc pediatric transplantation, cold ischemia time ranged from 14 to 26.2 hours (mean, 21.3 hours). Mean serum creatinine at 3, 6, and 12 months after transplantation was 1.53 +/- 0.57, 1.27 +/- 0.27, and 1.15 +/- 0.26 mg/dL compared with 1.93 +/- 1.35, 1.81 +/- 1.17, and 1.73 +/- 0.85 (P = .08) in recipients of single kidneys from ideal cadaveric donors (UNOS criteria, n = 368). Patient and graft survival at 1 year were 88.8% compared with 91.2% and 85% with ideal donors (P = NS), respectively. Three cases required additional surgery. There was one death due to a cerebral vascular accident. CONCLUSION: The present study confirmed the excellent results achieved with transplantation using en bloc kidneys from young donors.

Stage-specific gene expression during Trypanosoma cruzi metacyclogenesis

The process of Trypanosoma cruzi metacyclogenesis involves the transformation of noninfective epimastigotes into metacyclic trypomastigotes, which are the pathogenic form. The analysis of stage-specific genes during T. cruzi metacyclogenesis may provide insight into the mechanisms involved in the regulation of gene expression in trypanosomatids. It may also improve the understanding of the mechanisms responsible for the pathology of Chagas disease, and could lead to the identification of new targets for chemotherapy of this disease. We have demonstrated that during metacyclogenesis the expression of several genes is controlled at the translational level by an alternative regulatory mechanism. This mechanism may involve the mobilization of mRNA to the translation machinery. We have been using self-made T. cruzi microarrays to investigate the role of polysomal mobilization in modulating gene expression during metacyclogenesis

Trypanosoma cruzi: a gene family encoding chitin-binding-like proteins is posttranscriptionally regulated during metacyclogenesis.

The development of the representation of differential expression method has lead to the cloning of Trypanosoma cruzi stage-specific genes. We used this method to characterize a multicopy gene family differentially expressed during metacyclogenesis. The genomic and cDNA clones sequenced encoded three short cysteine-rich polypeptides, of two types, with predicted molecular masses of 7.1, 10.4, and 10.8 kDa. We searched GenBank for similar sequences and found that the sequences of these clones were similar to that encoding the wheat germ agglutinin protein. The region of similarity corresponds to the chitin-binding domain, with eight similarly positioned half-cysteines and conserved aromatic residues involved in chitin recognition. Multiple copies of the genes of this family are present on a high- molecular-mass chromosome. We studied the expression of genes of this family during metacyclogenesis by determining messenger RNA (mRNA) levels. The mRNAs for the members of this gene family were present in the total RNA fraction but were mobilized to the polysomal fraction of adhered (differentiating) epimastigotes during metacyclogenesis, with a peak of accumulation at 24 of differentiation. Polyclonal antisera were raised against a recombinant protein and a synthetic peptide. The specific sera obtained detected 7- and 11-kDa proteins in T. cruzi total protein extracts. The 11-kDa protein was present in similar amounts in the various cell populations, whereas the 7-kDa protein displayed differential synthesis during metacyclogenesis, with maximal levels in 24-h-adhered (differentiating) epimastigotes.

Cloning and characterization of the metacyclogenin gene, which is specifically expressed during Trypanosoma cruzi metacyclogenesis.

We isolated a gene that is differentially expressed during Trypanosoma cruzi metacyclogenesis by the representation of differential expression (RDE) method, using differentiating epimastigotes cultured in chemically defined medium. This gene, the metacyclogenin gene, encodes a 630-nucleotide mRNA that is specifically associated with the polysomes of epimastigotes allowed to differentiate for 24 h. We sequenced and characterized the metacyclogenin gene and found that there were at least three copies of the gene organized into tandem 2.8 kb repeats in the genome of T. cruzi Dm28c. We analyzed the repeats and found that they contained two other genes, one encoding tryparedoxin peroxidase and the other encoding a 0.6 kb mRNA (named associated gene or AG) with sequences showing no significant similarity to those in the GenBank database. Northern blot analysis of polysomal RNA extracted from replicating and differentiating epimastigotes showed that metacyclogenin and AG genes displayed similar patterns of expression. Their products were detected only in differentiating epimastigotes, whereas tryparedoxin peroxidase was detected only in the polysomal RNA fraction of replicating and differentiating epimastigotes. In Northern blots of total RNA from differentiating and replicating epimastigotes, the genes studied were detected in both cell populations. The differential expression of the metacyclogenin gene was confirmed by immunocytochemistry studies showing that the protein is detected only in differentiating (adhered) epimastigote. The results suggest that mRNA mobilization to polysomes is an important mechanism in the regulation of gene expression in T. cruzi.

Differential gene expression during Trypanosoma cruzi metacyclogenesis.

The transformation of epimastigotes into metacyclic trypomastigotes involves changes in the pattern of expressed genes, resulting in important morphological and functional differences between these developmental forms of Trypanosoma cruzi. In order to identify and characterize genes involved in triggering the metacyclogenesis process and in conferring to metacyclic trypomastigotes their stage specific biological properties, we have developed a method allowing the isolation of genes specifically expressed when comparing two close related cell populations (representation of differential expression or RDE). The method is based on the PCR amplification of gene sequences selected by hybridizing and subtracting the populations in such a way that after some cycles of hybridization-amplification genes specific to a given population are highly enriched. The use of this method in the analysis of differential gene expression during T. cruzi metacyclogenesis (6 hr and 24 hr of differentiation and metacyclic trypomastigotes) resulted in the isolation of several clones from each time point. Northern blot analysis showed that some genes are transiently expressed (6 hr and 24 hr differentiating cells), while others are present in differentiating cells and in metacyclic trypomastigotes. Nucleotide sequencing of six clones characterized so far showed that they do not display any homology to gene sequences available in the GeneBank.