L-2-hydroxyglutaric Aciduria in Two Female Yorkshire Terriers

Two female Yorkshire terrier puppies were presented with generalized tonic-clonic seizures and ataxia. MRI revealed bilaterally symmetrical, diffuse regions of gray matter hyperintensity on T2-weighted and fluid-attenuated inversion recovery sequences. Urinary organic acids were quantified by gas chromatography-mass spectroscopy and were consistent with a diagnosis of L-2-hydroxyglutaric aciduria (L2HGA). The L2HGDH gene encodes for the enzyme L-2-hydroxyglutarate dehydrogenase, which helps break down L-2-hydroxyglutaric acid. In both puppies described in this report, a homozygous mutation at the translation initiation codon of the homolog canine L2HGDH gene was detected (c.1A>G; p.Met1?), confirming the diagnosis of L2HGA at the DNA level. Canine L2HGA is caused by more than one mutation of L2HGDH, as reported in humans.     

Effects of protein source and lipid supplementation on conservation and feed value of total mixed ration silages for finishing beef cattle

The objective of this study was to examine the conservation process and feed value of total mixed ration (TMR) silages. In exp. 1, we evaluated the fermentation pattern and aerobic stability of TMR silages containing different protein and lipid supplementations. In exp. 2, we compared the performance of finishing beef heifers fed those TMR silages. In both experiments, treatments were as follows: ensiled TMR with urea (U); ensiled TMR without a protein supplement at ensiling, but soybean meal supplemented at feeding to balance diet crude protein (CP) in exp. 2 (SMnf; where the acronym nf indicates nonfermented); ensiled TMR with soybean meal (SM); and ensiled TMR with rolled soybean grain (SG). Thirty-two Nellore heifers (313 ± 8.8 kg shrunk body weight [SBW]) were blocked by initial SBW, housed in individual pens, and enrolled in exp. 2 for 82 d. In exp. 1, treatment without a protein supplement (SMnf) had a lower content of CP, soluble CP, NH₃-N, pH, and Clostridium count compared with U (P ≤ 0.03). Lactic acid concentrations tended to be reduced for SMnf compared with U (P = 0.09). Ethanol concentration was reduced in SG compared with SM (P < 0.01). 1,2-Propanediol concentration was increased in SMnf compared with U (P < 0.01), reduced in SM compared with SMnf (P = 0.02), and increased in SG compared with SM (P = 0.02). Dry matter (DM) loss during fermentation was low and similar among treatments (~3.7%). All silages remained stable during 10 d of aerobic exposure after feed out. Considering fermentation traits, such as pH (≤4.72), NH₃-N (<10% of N, except for U treatment), butyric acid (<0.05 % DM), and DM losses (<3.70% DM), all silages can be considered well conserved. In exp. 2, diets were isonitrogenous because soybean meal was added to SMnf before feeding. Compared with SM, cattle fed SG made more meals per day (P = 0.04) and tended to have a decreased intermeal interval (P = 0.09). DM intake, average daily gain, final SBW, hot carcass weight, Biceps femoris fat thickness, and serum levels of triglycerides and cholesterol were increased for SG compared with SM (P ≤ 0.05). In brief, TMR silages exhibited an adequate fermentation pattern and high aerobic stability. The supplementation of true protein did not improve animal performance, whereas the addition of soybean grain as a lipid source improved the performance of finishing cattle fed TMR silages.     

Virulence factors and genetic variability of uropathogenic Escherichia coli isolated from dogs and cats in Italy

In this study, the association between virulence genotypes and phylogenetic groups among Escherichia (E.) coli isolates obtained from pet dogs and cats with cystitis was detected, and fingerprinting methods were used to explore the relationship among strains. Forty uropathogenic E. coli (UPEC) isolated from dogs (n = 30) and cats (n = 10) in Italy were analysed by polymerase chain reaction (PCR) for the presence of virulence factors and their classification into phylogenetic groups. The same strains were characterized by repetitive extragenic palindromic (REP)- and enterobacterial repetitive intergenic consensus (ERIC)-PCR techniques. We found a high number of virulence factors such as fimbriae A, S fimbriae (sfa) and cytotoxic necrotizing factor 1 (cnf1) significantly associated with phylogenetic group B2. We demonstrated a high correlation between α-hemolysin A and pyelonephritis C, sfa, and cnf1 operons, confirming the presence of pathogenicity islands in these strains. In addition, UPEC belonging to group B2 harboured a greater number of virulence factors than strains from phylogenetic groups A, B1, and D. REP- and ERIC-PCR grouped the UPEC isolates into two major clusters, the former grouping E. coli strains belonging to phylogenetic group B2 and D, the latter grouping those belonging to groups A and B1. Given the significant genetic variability among the UPEC strains found in our study, it can be hypothesized that no specific genotype is responsible for cystitis in cats or dogs.     

Antigenicity of a whole parasite vaccine as promising candidate against canine leishmaniasis

Human visceral leishmaniasis, one of the most important zoonoses, is caused by the protozoa Leishmania chagasi (syn. L. infantum) and is present as a fatal disease common in South America and Europe where dogs and wild canids are the main reservoirs. A vaccine against visceral leishmaniasis would be an important tool in the control of this disease in dogs. Although the current strategies for vaccination against leishmaniasis are based on the use of recombinant antigens, killed vaccines are still attractive in terms of stability of their biochemical composition and antigenicity, cost, and safety. Here we evaluate the immunogenicity of a whole parasite vaccine as a promising candidate against canine leishmaniasis, demonstrated by cellular reactivity, changes in the cellular profile of the peripheral blood and by the differential production of immunoglobulins. Our results showed that immunization elicited mainly a strong cellular reactivity and increase in T-lymphocytes, particularly the subpopulation CD8(+) that would be related to the control of tissue parasitism. In addition, a higher production of anti-Leishmania total IgG, characterized by mixed isotypes profile (IgG1 and IgG2), was demonstrated.     

Thromboelastography results on citrated whole blood from clinically healthy cats depend on modes of activation

Background

During the last decade, thromboelastography (TEG) has gained increasing acceptance as a diagnostic test in veterinary medicine for evaluation of haemostasis in dogs, however the use of TEG in cats has to date only been described in one previous study and a few abstracts. The objective of the present study was to evaluate and compare three different TEG assays in healthy cats, in order to establish which assay may be best suited for TEG analyses in cats.

Methods

90 TEG analyses were performed on citrated whole blood samples from 15 clinically healthy cats using assays without activator (native) or with human recombinant tissue factor (TF) or kaolin as activators. Results for reaction time (R), clotting time (K), angle (α), maximum amplitude (MA) and clot lysis (LY30; LY60) were recorded.

Results

Coefficients of variation (CVs) were highest in the native assay and comparable in TF and kaolin activated assays. Significant differences were observed between native and kaolin assays for all measured parameters, between kaolin and TF for all measured parameters except LY60 and between native and TF assays for R and K.

Conclusion

The results indicate that TEG is a reproducible method for evaluation of haemostasis in clinically healthy cats. However, the three assays cannot be used interchangeably and the kaolin- and TF activated assays have the lowest analytical variation indicating that using an activator may be superior for performing TEG in cats.     

A rapid,inexpensive, and semi-quantitative method for determining pollen tube extension using fluorescence

Background

To identify plant genes involved in various key traits, QTL mapping is a powerful approach. This approach is based on the use of mapped molecular markers to identify genomic regions controlling quantitative traits followed by a fine mapping and eventually positional cloning of candidate genes. Mapping technologies using SNP markers are still rather expensive and not feasible in every laboratory. In contrast, microsatellite (also called SSR for Simple Sequence Repeat) markers are technologically less demanding and less costly for any laboratory interested in genetic mapping.

Results

In this study, we present the development and the characterization of a panel of 96 highly polymorphic SSR markers along the Arabidopsis thaliana genome allowing QTL mapping among accessions of the Versailles 24 core collection that covers a high percentage of the A. thaliana genetic diversity. These markers can be used for any QTL mapping analysis involving any of these accessions. We optimized the use of these markers in order to reveal polymorphism using standard PCR conditions and agarose gel electrophoresis. In addition, we showed that the use of only three of these markers allows differentiating all 24 accessions which makes this set of markers a powerful tool to control accession identity or any cross between any of these accessions.

Conclusion

The set of SSR markers developed in this study provides a simple and efficient tool for any laboratory focusing on QTL mapping in A. thaliana and a simple means to control seed stock or crosses between accessions.