Rumen odd and branched chain fatty acids in relation to in vitro rumen volatile fatty acid productions and dietary characteristics of incubated substrates

A first aim of this batch in vitro experiment (21 h) was to use changes in odd and branched chain fatty acid (OBCFA) patterns to suggest shifts in microbial populations, associated with four types of incubated whole dairy cow diets. Principal component analysis suggested higher dietary starch increased the proportion of C15:0 and C17:0, whereas increased neutral detergent fibre content was positively related to anteiso C15:0 concentrations, which is in agreement with the importance of these fatty acids in respectively amylolytic and cellulolytic bacteria. A second aim of the experiment was to relate rumen volatile fatty acid proportions to OBCFA by principal component regression and to compare these relations with predictions based on diet proximate composition. The R2 values achieved for the regressions between acetate, propionate and butyrate, and OBCFA were 79.6%, 86.6% and 84.9% respectively. Moreover, in the current study, predictions of the rumen fermentation pattern showed higher R2 (p < 0.01) when based on OBCFA compared with proximate feed composition. If relations persist in vivo, there could be scope for milk OBCFA to predict the supply of specific rumen nutrients.     

Exploration of plant genomes in the FLAGdb++ environment

Background

Efficient high throughput screening systems of useful mutants are prerequisite for study of plant functional genomics and lots of application fields. Advance in such screening tools, thanks to the development of analytic instruments. Direct analysis in real-time (DART)-mass spectrometry (MS) by ionization of complex materials at atmospheric pressure is a rapid, simple, high-resolution analytical technique. Here we describe a rapid, simple method for the genetic discrimination of intact Arabidopsis thaliana mutant seeds using metabolic profiling by DART-MS.

Results

To determine whether this DART-MS combined by multivariate analysis can perform genetic discrimination based on global metabolic profiling, intact Arabidopsis thaliana mutant seeds were subjected to DART-MS without any sample preparation. Partial least squares-discriminant analysis (PLS-DA) of DART-MS spectral data from intact seeds classified 14 different lines of seeds into two distinct groups: Columbia (Col-0) and Landsberg erecta (Ler) ecotype backgrounds. A hierarchical dendrogram based on partial least squares-discriminant analysis (PLS-DA) subdivided the Col-0 ecotype into two groups: mutant lines harboring defects in the phenylpropanoid biosynthetic pathway and mutants without these defects. These results indicated that metabolic profiling with DART-MS could discriminate intact Arabidopsis seeds at least ecotype level and metabolic pathway level within same ecotype.

Conclusion

The described DART-MS combined by multivariate analysis allows for rapid screening and metabolic characterization of lots of Arabidopsis mutant seeds without complex metabolic preparation steps. Moreover, potential novel metabolic markers can be detected and used to clarify the genetic relationship between Arabidopsis cultivars. Furthermore this technique can be applied to predict the novel gene function of metabolic mutants regardless of morphological phenotypes.     

Biometrical, biological, biochemical and molecular characteristics of Meloidogyne incognita isolates and related species

Meloidogyne incognita is one of the most polyphagous species of root-knot nematodes occurring in Brazil and worldwide. Eight M. incognita isolates were studied, representing two enzymatic phenotypes (esterase and malate desydrogenase: I1/N1, I2/N1) and four cryptic Meloidogyne sp.1 (S2/N1) isolates, representing one cytological type (3n?=?40–46). Three M. hispanica isolates (Hi3/N1, 2n?=?32–36) and two of an atypical Meloidogyne sp.2 (S2a/N3, 3n?=?40–44) were included in this study for comparison. All isolates were tested with three M. incognita-specific molecular markers. The primer pairs B06F/R, miF/R and incK14F/R amplified three species-specific fragments of 1,200?bp, 955?bp and 399?bp, respectively for M. incognita and Meloidogyne sp.1 isolates. No amplification occurred in the M. hispanica and Meloidogyne sp.2 isolates, except with primers miF/R (1,650?bp). The genetic variability of the Meloidogyne spp. isolates was evaluated, using RAPD and ISSR markers. The phylogenetic analyses revealed two strongly supported monophyletic clades: clade I, consisting of M. hispanica and the atypical Meloidogyne sp.2 isolates, and clade II, clustering together all M. incognita and the Meloidogyne sp.1 isolates. Considering the biometrical, cytological and molecular approaches, it was possible to conclude that the isolates with three enzymatic phenotypes (I1/N1, I2/N1 and S2/N1) presented the characteristics described for M. incognita. Some correlations were detected between the isozymatic phenotypes and the tree topology (S2a/N3, Hi3/N1, I1/N1, S2/N1), but no strict correlation could be observed for the phenotype I2/N1 and one isolate of S2/N1. Morphologically, the Msp.2 isolates differ from M. incognita and M. hispanica by the female stylet features presenting straight cone tip and round pear shaped knobs, posteriorly sloping. The results of this study suggested that the Msp.2 isolates with phenotypes S2aN3 belong to a new or an unidentified species closely related to M. hispanica.     

Mutations in the juxtamembrane domain of c-KIT are associated with higher grade mast cell tumors in dogs

Mast cell tumors are among the most commonly seen tumors of the skin in dogs and are more highly aggressive than mast cell tumors of other species. Some breeds display a markedly higher incidence of mast cell tumor development than others and appear to have some genetic predisposition. Recently, mutations have been found in canine mast cell tumor tissues and cell lines within the juxtamembrane domain of the protooncogene c-KIT In previous studies utilizing a small number of cases, no association between the presence of a mutation and the breed of dog or grade of the tumor could be identified. An expanded study with a larger sample set was performed to explore this possibility. The juxtamembrane domain of c-KIT was amplified using the polymerase chain reaction from genomic DNA preparations of 88 paraffin-embedded mast cell tumors from selected breeds. Mutations, consisting of duplications and deletions, were found in 12 of the tumors. A significant association was found between the presence of a mutation and a higher grade of tumor but not between breed and grade or between breed and the presence of a mutation.     

Detection of enzootic nasal tumor virus (ENTV) in a sheep flock in southern Brazil

Enzootic nasal tumor (ENT) is a contagious neoplasm associated with enzootic nasal tumor virus (ENTV), which may induce disease in sheep (ENTV-1) and goats (ENTV-2). This study aimed to describe the occurrence of ENT in two Texel sheep (Ovis aries) from a 75-sheep flock, located in the city of Gravataí, southern Brazil. Animals used to be purchased from different origins, and no specific tests for disease monitoring or quarantine procedure were performed. Affected animals presented respiratory distress, anorexia with severe weight loss, and mucopurulent unilateral nasal discharge. Necropsy was performed in both animals and nasal cavity masses were observed. Histopathological analysis demonstrated an epithelial neoplasm compatible with nasal adenocarcinoma. PCR using a protocol that amplifies a 591 bp sequence of 5’LTR-gag region of ENTV1 was performed followed by DNA sequencing. Both samples were positive, and the sequences obtained presented highest identity (97%) with ENTV strain TN28 (GenBank accession number MH899613) detected in a Texel sheep from Scotland. This is the first report of ENTV-1 leading to enzootic nasal tumor in sheep in Latin America, which confirms the presence of the retrovirus in sheep flocks in the Brazilian territory.

     

A three‐dimensional cell culture system as an in vitro canine mammary carcinoma model for the expression of connective tissue modulators

In this study, derived complex carcinoma (CC) and simple carcinoma (SC) cell lines were established and cultured under two‐dimensional (2D) and three‐dimensional (3D) conditions. The 3D was performed in six‐well AlgiMatrix? (LifeTechnologies®, Carlsbad, CA, USA) scaffolds, resulting in spheroids sized 50–125 µm for CC and 175–200 µm for SC. Cell viability was demonstrated up to 14 days for both models. Epidermal growth factor receptor (EGFR) was expressed in CC and SC in both systems. However, higher mRNA and protein levels were observed in SC 2D and 3D systems when compared with CC (P < 0.005). The connective tissue modulators, metalloproteinases‐1, ‐2, ‐9 and ‐13 (MMPs), relaxin receptors 1 and 2 (RXR1 and RXR2) and E‐cadherin (CDH1) were quantitated. All were upregulated similarly when canine mammary tumour (CMT)‐derived cell lines were cultured under 3D AlgiMatrix, except CDH1 that was downregulated (P < 0.005). These results are promising towards the used of 3D system to increase a high throughput in vitro canine tumour model.     

Assessment of the efficacy of composite surgery for the treatment of dorsal displacement of the soft palate in a group of 53 racing Thoroughbreds (1990-1996)

REASONS FOR PERFORMING STUDY: There has been no objectively assessed case-control study of the efficacy of surgery to correct dorsal displacement of the soft palate (DDSP) previously reported. HYPOTHESIS: Composite surgery has a beneficial result on racing performance in horses affected with DDSP as compared to a matched control population. METHODS: Race records were obtained for 53 racing Thoroughbreds which underwent composite staphylectomy, sternothyrohyoideus myectomy and ventriculectomy for correction of idiopathic DDSP at the University of Bristol between 1990 and 1996. Each surgical case was matched for age, sex and training yard with 2 control horses. The racing performance, based on prize money won, of surgical cases and control horses were compared for 3 races run before and after the date of surgery. RESULTS: Ninety-two percent of the surgical cases returned to racing after surgery. There was a significant increase in earnings of the surgical group before and after surgery (P = 0.011), but there was no significant difference in earnings of the control group before and after the date of surgery (P = 0335). Sixty percent of the surgical group had higher earnings after surgery than before, compared to 40% of controls. When horses which underwent surgery were ranked relative to their 2 matched controls, surgical cases did not significantly change in rank (P = 033), whereas control horses significantly decreased in rank (P = 0.012). Additionally, horses within the surgical group were more likely (P < 0.01) to start in 3 post operative races than those in the control group. CONCLUSIONS AND POTENTIAL RELEVANCE: Composite surgery had a beneficial effect on racing performance of horses afflicted with idiopathic DDSP, and further studies to evaluate objectively the usefulness of other surgical techniques are warranted.     

Higher feeding diets effects on age and liveweight gain at puberty in crossbred Nelore × Hereford heifers

This study was conducted to determine the age and liveweight at puberty of 120 crossbred beef heifers submitted to four diets to achieve predetermined weight gains (kg/day): 0.5 (G500; n?=?32), 0.75 (G750; n?=?32), 1.0 (G1000; n?=?29), and 1.25 (G1250; n?=?27). Animals were classified depending on their level of crossing between Nelore (N) and Hereford (H): 25%N-75%H, 37.5%N-63.5%H, 43.7%N-56.7%H, 50%N-50%H, and 75%N-25%H. Reproductive evaluation was performed at the beginning and at the end of the experimental period by ultrasonography and hormone analysis. The average age and liveweight at puberty were 388.0?±?1.9 days and 331.4?±?1.3 kg, respectively. Animals from the 25%N-75%H group reached puberty earlier than heifers from other genetic groups supplemented with G1250 diet (P?<?0.05). Heifers with higher degree of Nelore (75%N-25%H) fed with G1000 diet showed estrus 42 days prior to mating, but only 57 % reached puberty at mating (P?<?0.05). Heifers with follicles of higher diameter reached puberty (P?<?0.05) earlier. Higher average daily weight gain showed a positive effect on follicular diameter and IGF-I level at puberty (P?<?0.01). Concentrations of GH were lower in heifers fed G1250 compared to G1000 diet (P?<?0.05). There was a significant interaction between nutritional level and insulin levels at puberty (P?<?0.01). We demonstrated the relationship between IGF-I and average daily gain on the onset of puberty in heifers. In conclusion, heifers submitted to the higher feeding level showed a higher follicular diameter and were younger at puberty.     

Comparison between Haemophilus parasuis infection in colostrums-deprived and sow-reared piglets

The aim of this study was to compare the development of Glasser's disease in sow-reared and colostrum-deprived piglets. Ninety piglets from a commercial pig farm in Spain were used. The farm was positive for Haemophilus parasuis. Fifty-two pigs were sow-reared (SR) and 38 were colostrum-deprived (CD) piglets. The animals were intratracheally inoculated with H. parasuis serovar 5 and sacrificed at 1, 2 and 3 days post-infection. To assess the development of disease, antibody titers, clinical signs, pathological lesions, microbiological isolation and PCR amplification were compared between the groups. Inoculation of SR pigs did not cause clinical signs or lesions of Glasser's disease. In SR pigs, H. parasuis isolation and specific PCR amplification from tissues showed a very low number of positive samples. In contrast, in CD pigs, inoculation resulted in the typical signs and lesions of Glasser's disease. Positive microbiological isolation and specific PCR products were obtained from the majority of the tissues tested, and no antibodies against H. parasuis were detected. The experimental infection using CD pigs describes a successful method to study this microorganism and confirms the important role that maternal antibodies play in protection against clinical signs and disease.