Combined use of gliadins and SSRs to analyse the genetic variability of the Spanish collection of cultivated diploid wheat (Triticum monococcum L. ssp. monococcum)

This work studied the combined use of gliadins and SSRs to analyse inter- and intra-accession variability of the Spanish collection of cultivated einkorn (Triticum monococcum L. ssp. monococcum) maintained at the CRF-INIA. In general, gliadin loci presented higher discrimination power than SSRs, reflecting the high variability of the gliadins. The loci on chromosome 6A were the most polymorphic with similar PIC values for both marker systems, showing that these markers are very useful for genetic variability studies in wheat. The gliadin results indicated that the Spanish einkorn collection possessed high genetic diversity, being the differentiation large between varieties and small within them. Some associations between gliadin alleles and geographical and agro-morphological data were found. Agro-morphological relations were also observed in the clusters of the SSRs dendrogram. A high concordance was found between gliadins and SSRs for genotype identification. In addition, both systems provide complementary information to resolve the different cases of intra-accession variability not detected at the agro-morphological level, and to identify separately all the genotypes analysed. The combined use of both genetic markers is an excellent tool for genetic resource evaluation in addition to agro-morphological evaluation.     

Influence of fruit ripeness at the time of seed extraction on pepper (Capsicum annuum) seed germination

The influence of fruit ripeness (half ripe, fully ripe, overripe) at the time of seed extraction on seed germination behaviour, at 25 °C and 13 °C, was studied in two Spanish pepper cultivars for canning. Seeds from half ripe fruits had a poorer behaviour than those taken from fully ripe fruits, especially at 13 °C. Room ripening and overripening of half ripe fruits improved the germination behaviour of the seeds although this behaviour was poorer than that of seeds extracted from fully ripe fruits that were allowed to overripen. Overripening of fully ripe picked fruits slightly improved germination behaviour.     

Avascular necrosis of the femoral heads in a red panda (Ailurus fulgens fulgens): possible Legg-Calve-Perthes disease.

A 17-mo-old captive-born female red panda (Ailurus fulgens fulgens) presented with a sudden onset of lameness in its left hind leg was diagnosed radiographically as having possible severe, bilateral Legg-Calve-Perthes disease with fracture of the great trochanter of the left femur. Surgical repair of the fracture was performed using pins and a tension band wire through a lateral approach to the hip. This is the first case reported at Madrid Zoo-Aquarium, where 63 individuals have been bred over 15 yr.     

In vitro micrografting and its applications to fruit science

Improvements have been brought to in vitro micrografting. The breaking of dormancy by dipping the seeds of the rootstock in concentrated solutions of benzylaminopurine avoids having recourse to long stratification in the cold (60–120 days according to species). The initial pre-treatment of apices with cytokinin allows their development and facilitates later grafting. Moreover, adding DIECA to the cut sections avoids oxidation and, together with the insertion of a piece of agar between apex and rootstock, increases the success of micrografting. Finally, a special technique of in vivo micrografting of pretreated apices on young plants has been developed.Micrografting is commonly used to obtain peach trees free of Sharka and NRS viruses, and to study incompatibility mechanisms in heterografts, leading to an early diagnosis of incompatibilities.     

Pathophysiology of hepatic coccidiosis in rabbits

Four groups of 5 40-day-old, Eimeria stiedai-naive, New Zealand rabbits were infected with 0 (Group A), 10² (B), 10³ (C) and 10⁴ (D) sporulated oocysts of E. stiedai and observed for 50 days. Serum glutamic pyruvic (GPT) and glutamic oxalacetic (GOT) transaminases, bilirubinemia, lipemia, proteinemia, glycemia, oocyst output, body, carcass and liver weights, and mortality were recorded. Four physiopathological events were identified: (1) a phase of indirect damage to the hepatocytes that takes place during the first 2 weeks of infection and is characterized by increased transaminases; (2) a cholestatic period consequent to the production of oocysts that begins suddenly in the 3rd week, diminishes gradually towards the 7th week, and is characterized by a rise of bilirubinemia and lipemia; (3) a stage of metabolic dysfunction that begins in the 3rd–4th week, intensifies for the next 3 weeks and starts to recover during the 7th week. It is characterized by hypoproteinemia and hypoglycemia; (4) a period of immunodepression characterized by the inability of the heavily infected host to inhibit oocyst production. Apart from the cholestatic phase, the respective pathogenic mechanisms remains to be studied.     

Effects of hemolysis, lipemia, hyperbilirrubinemia, and anticoagulants in canine C-reactive protein, serum amyloid A, and ceruloplasmin assays

The objective of the present study was to determine the effects that hemolysis, lipemia, bilirubinemia, and anticoagulants might have on the most commonly used assays for C-reactive protein and serum amyloid A, and determination of ceruloplasmin values in dogs. Solutions of hemoglobin, lipid, and bilirubin were added to serum aliquots. Additionally, serum and plasma samples with different anticoagulants (heparin, EDTA, and citrate) were obtained from healthy dogs. Hemolysis, lipemia, and hyperbilirubinemia interfered significantly with the C-reactive protein and ceruloplasmin results, but not with those for the serum amyloid A assay. The use of anticoagulants produced significant changes in the results for the assays tested. However, the magnitude of the differences caused by the interfering substances does not appear to have an important impact on the clinical interpretation of the tests.     

Participation of Rhipicephalus sanguineus (Acari: Ixodidae) in the epidemiology of canine visceral leishmaniasis

The vectorial competence of the tick Rhipicephalus sanguineus is discussed in relation to the epidemiology of canine visceral leishmaniasis, taking into account its strict association with dogs and the low indices of natural infection presented by its known vector, the phlebotomine sand fly Lutzomyia longipalpis. In order to evaluate natural infection by Leishmania chagasi and the infectivity of these parasites in the tick, 39 specimens (6 females, 11 males and 22 nymphs) of R. sanguineus were removed from 21 dogs showing diverse symptoms of zoonotic visceral leishmaniasis (ZVL). Six ticks (15.4%) gave positive results for the genus Leishmania using the PCR technique. To determine the infectivity of the parasites, 36 hamsters were inoculated orally and peritoneally with macerates of ticks removed from nine dogs symptomatic for visceral leishmaniasis. After 6 months the hamsters were sacrificed and necropsied. Serum was removed for IFAT, as well as spleen and liver fragments to make imprint smears and for PCR. Eight (88.9%) of these dogs presented ticks that were infective for 14 hamsters (41.2%), 12 (85.7%) of them infected peritoneally and two (14.3%) orally. PCR revealed 27 smears (40.9%) to be positive, 20 (62.5%) of them infected peritoneally and seven (20.6%) orally. IFAT showed 14 positive animals (41.2%). Based on these findings, we suggest that the vectorial capacity of R. sanguineus for L. chagasi should be evaluated further, opening new perspectives in the epidemiology of ZVL.     

Experimental use of polyamide bands in combination with intramedullary pinning for repair of oblique femoral fractures in rabbits

OBJECTIVE: To evaluate the use of polyamide bands, manufactured for securing electrical cables, for repair of oblique femoral fractures in rabbits. STUDY DESIGN: Experimental study. ANIMALS: Twelve nulliparous, 21-25-week-old, California female rabbits. METHODS: Rabbits were divided into 3 groups (n=4) and studied for 14, 28, or 56 days. A Z-shaped mid-diaphyseal femoral osteotomy was repaired with a 2.5 mm intramedullary pin and a polyamide 6.6 cerclage band. Healing was evaluated at intervals by physical examination, limb use, femoral radiographs, and callus histology. RESULTS: Rabbits had early limb use with good wound healing. From the 1st day, movement of the hip and stifle joints was satisfactory. Radiographically, fractures were healed at 28 days. Histologically, there was no foreign body reaction and bone healing was normal. CONCLUSION: Nylon cerclage band application was accomplished easily, maintained reduction, and resulted in good healing and limb use. CLINICAL RELEVANCE: Sterilized nylon bands, manufactured for electrical use, can be used for cerclage in rabbits.     

Minimally invasive patent ductus arteriosus occlusion in 5 dogs

OBJECTIVE: To report a technique for minimally invasive occlusion of patent ductus arteriosus (PDA) and outcome in 5 dogs. STUDY DESIGN: Clinical cases. Animals: Five, 4-6-month-old, dogs with PDA. MATERIALS AND METHODS: Titanium ligating clips were used for PDA closure in all dogs. Three dogs had video-enhanced mini-thoracotomy PDA occlusion. Two other dogs had thoracoscopic PDA occlusion using a custom-designed thoracoscopy clip applicator. RESULTS: Thoracoscopic PDA occlusion was successful in both dogs in which it was attempted. Complete PDA closure was achieved in 4 dogs. Three months after surgery, the largest dog had residual ductal flow that hemodynamically was insignificant. CONCLUSIONS: Although technically demanding, minimally invasive PDA occlusion is a safe and reliable technique in dogs. Preoperative measurement of the diameter of the PDA is crucial to determine if complete closure with metal clips can be achieved. CLINICAL RELEVANCE: Minimally invasive PDA occlusion should be considered as an alternative to occlusion via conventional thoracotomy.     

Mapping of B-cell epitopes in the N-terminal repeated peptides of Anaplasma marginale major surface protein 1a and characterization of the humoral immune response of cattle immunized with recombinant and whole organism antigens

Major surface protein (MSP) 1a of the genus type species Anaplasma marginale (Rickettsiales: Anaplasmataceae) together with MSP1b forms the MSP1 complex. MSP1a has been shown to be involved in adhesion, infection and tick transmission of A. marginale, as well as to contribute to protective immunity in cattle. A differential antibody response to MSP1a and MSP1b was observed in cattle immunized with A. marginale derived from bovine erythrocytes (anti-MSP1a response) or cultured tick cells (anti-MSP1b response). In this study, we further characterized the MSP1a antibody response of cattle using several immunogens, including recombinant MSP1a (rMSP1a) protein, erythrocyte- or tick cell culture-derived A. marginale, or a combination of tick cell culture-derived A. marginale and rMSP1a. The MSP1a antibody response to all these immunogens was directed primarily against the N-terminal region of MSP1a that contains tandemly repeated peptides, whereas low antibody levels were detected against the C-terminal portion. Linear B-cell epitopes of MSP1a were mapped using synthetic peptides representing the entire sequence of the protein that were prepared by SPOT synthesis technology. Only two peptides in the N-terminal repeats were recognized by sera from immunized cattle. These peptides shared the sequence SSAGGQQQESS, which is likely to contain the linear B-cell epitope that was recognized by the pools of bovine sera. The average differential of antibody titers against MSP1a minus those against MSP1b correlated with lower percent reductions in PCV. A preferential antibody response to MSP1a was observed in cattle immunized with erythrocyte-derived, cell culture-derived plus rMSP1a or rMSP1a alone, and the percent reduction PCV was significantly lower in these cattle as compared with the other immunization groups. These results provide insight into the bovine antibody response against A. marginale and the role of MSP1a in protection of cattle against A. marginale infection.