Anatomía del Desarrollo del Tracto Digestivo y Forma Corporal Externa Durante el Período Embrionario en el Caprino (Copra hircus)

Anatomy of the digestive tract and external body development during the embryonic stages of the caprine (Copra, bircus) The purpose of this work has been to establish the pattern of prenatal growth and normal development of the digestive tract and annex glands during goat embryonic stages. 21 embryos with ages ranging from 14 to 34 days (1.69 to 5.90 cm CR) as determined by registering the mating time, were obtained by cesarea. This material was histologically processed to obtain complete serial sections of the stomatodaeum, foregut, midgut, hindgut and cloaca. In this work, it is chronologically described the morphogenesis and histogenesis of the mouth, hypophysis, pharynx and its derivatives, stomach, intestine, liver, pancreas and cloaca. The results obtained establish chronological comparisons with the development of the lamb and gives information on the unitary origin of the gastric compartments in ruminants.     

Major outer membrane proteins of Brucella spp.: past,present and future

The major outer membrane proteins (OMPs) of Brucella spp. were initially identified in the early 1980s and characterised as potential immunogenic and protective antigens. They were classified according to their apparent molecular mass as 36–38 kDa OMPs or group 2 porin proteins and 31–34 and 25–27 kDa OMPs which belong to the group 3 proteins. The genes encoding the group 2 porin proteins were identified in the late 1980s and consist of two genes, omp2a and omp2b, which are closely linked in the Brucella genome, and which share a great degree of identity (>85%). In the 1990s, two genes were identified coding for the group 3 proteins and were named omp25 and omp31. The predicted amino acid sequences of omp25 and omp31 share 34% identity. The recent release of the genome sequence of B. melitensis 16 M has revealed the presence of five additional gene products homologous to Omp25 and Omp31. The use of recombinant protein technology and monoclonal antibodies (MAbs) has shown that the major OMPs appear to be of little relevance as antigens in smooth (S) B. abortus or B. melitensis infections i.e. low or no protective activity in the mouse model of infection and low or no immunogenicity during host infection. However, group 3 proteins, in particular Omp31, appear as immunodominant antigen in the course of rough (R) B. ovis infection in rams and as important protective antigen in the B. ovis mouse model of infection. The major OMP genes display diversity and specific markers have been identified for Brucella species, biovars, and strains, including the recent marine mammal Brucella isolates for which new species names have been proposed. Recently, Omp25 has been shown to be involved in virulence of B. melitensis, B. abortus and B. ovis. Mutants lacking Omp25 are indeed attenuated in animal models of infection, and moreover provide levels of protection similar or better than currently used attenuated vaccine strain B. melitensis Rev.1. Therefore, these mutant strains appear interesting vaccine candidates for the future. The other group 3 proteins identified in the genome merit also further investigation related to the development of new vaccines.     

Clinical evaluation and prerelease management of American river otters in the second year of a reintroduction study

In the first year (1984) of a reintroduction study, 10 American river otters (Lutra canadensis) from Louisiana were transported to Oklahoma, held for 5 days for clinical evaluation, surgical implantation with intra-abdominal radiotelemetry devices, and then released in Oklahoma. Four of 10 otters released died within 32 days. Clinical evaluation indicated that respiratory tract disease, bacterial and parasitic infections, and inanition may have contributed to the death of these otters. In the second year (1985) of the study, an exotic feline diet was fed, and the holding period for 10 otters was increased to provide time for evaluation and treatment before surgery, postsurgical acclimation to Oklahoma, and reevaluation before release. Although the initial clinical findings on otters in the second year were similar to those found in the first year, otter body weights increased, and the prevalence and severity of clinical abnormalities decreased with treatment during the second-year holding period. Three of 10 second-year otters died during the holding period, and contributing causes of death were determined to be: trauma (hepatic hematoma), inanition, renal disease, pneumonia, salmonellosis (Salmonella anatum), and a retropharyngeal abscess (Klebsiella pneumoniae). Seven healthy otters were reintroduced into Oklahoma in 1985, and postrelease deaths were not experienced.     

Evaluation in mice of Brucella ovis attenuated mutants for use as live vaccines against B. ovis infection

Brucella ovis causes ram contagious epididymitis, a disease for which a specific vaccine is lacking. Attenuated Brucella melitensis Rev 1, used as vaccine against ovine and caprine brucellosis caused by B. melitensis, is also considered the best vaccine available for the prophylaxis of B. ovis infection, but its use for this purpose has serious drawbacks. In this work, two previously characterized B. ovis attenuated mutants (Δomp25d and Δomp22) were evaluated in mice, in comparison with B. melitensis Rev 1, as vaccines against B. ovis. Similarities, but also significant differences, were found regarding the immune response induced by the three vaccines. Mice vaccinated with the B. ovis mutants developed anti-B. ovis antibodies in serum of the IgG₁, IgG_2a and IgG_2b subclasses and their levels were higher than those observed in Rev 1-vaccinated mice. After an antigen stimulus with B. ovis cells, splenocytes obtained from all vaccinated mice secreted similar levels of TNF-α and IL12(p40) and remarkably high amounts of IFN-γ, a crucial cytokine in protective immunity against other Brucella species. By contrast, IL-1α -an enhancer of T cell responses to antigen- was present at higher levels in mice vaccinated with the B. ovis mutants, while IL-10, an anti-inflammatory cytokine, was significantly more abundant in Rev 1-vaccinated mice. Additionally, the B. ovis mutants showed appropriate persistence, limited splenomegaly and protective efficacy against B. ovis similar to that observed with B. melitensis Rev 1. These characteristics encourage their evaluation in the natural host as homologous vaccines for the specific prophylaxis of B. ovis infection.     

Mutants in the lipopolysaccharide of Brucella ovis are attenuated and protect against B. ovis infection in mice

Brucella spp. are Gram-negative bacteria that behave as facultative intracellular parasites of a variety of mammals. This genus includes smooth (S) and rough (R) species that carry S and R lipopolysaccharides (LPS), respectively. S-LPS is a virulence factor, and mutants affected in the S-LPS O-polysaccharide (R mutants), core oligosaccharide or both show attenuation. However, B. ovis is naturally R and is virulent in sheep. We studied the role of B. ovis LPS in virulence by mutating the orthologues of wadA, wadB and wadC, three genes known to encode LPS core glycosyltransferases in S brucellae. When mapped with antibodies to outer membrane proteins (Omps) and R-LPS, wadB and wadC mutants displayed defects in LPS structure and outer membrane topology but inactivation of wadA had little or no effect. Consistent with these observations, the wadB and wadC but not the wadA mutants were attenuated in mice. When tested as vaccines, the wadB and wadC mutants protected mice against B. ovis challenge. The results demonstrate that the LPS core is a structure essential for survival in vivo not only of S brucellae but also of a naturally R Brucella pathogenic species, and they confirm our previous hypothesis that the Brucella LPS core is a target for vaccine development. Since vaccine B. melitensis Rev 1 is S and thus interferes in serological testing for S brucellae, wadB mutant represents a candidate vaccine to be evaluated against B. ovis infection of sheep suitable for areas free of B. melitensis.     

Essential role of Fkbp6 in male fertility and homologous chromosome pairing in meiosis

Meiosis is a critical stage of gametogenesis in which alignment and synapsis of chromosomal pairs occur, allowing for the recombination of maternal and paternal genomes. Here we show that FK506 binding protein (Fkbp6) localizes to meiotic chromosome cores and regions of homologous chromosome synapsis. Targeted inactivation of Fkbp6 in mice results in aspermic males and the absence of normal pachytene spermatocytes. Moreover, we identified the deletion of Fkbp6 exon 8 as the causative mutation in spontaneously male sterile as/as mutant rats. Loss of Fkbp6 results in abnormal pairing and misalignments between homologous chromosomes, nonhomologous partner switches, and autosynapsis of X chromosome cores in meiotic spermatocytes. Fertility and meiosis are normal in Fkbp6 mutant females. Thus, Fkbp6 is a component of the synaptonemal complex essential for sex-specific fertility and for the fidelity of homologous chromosome pairing in meiosis.     

Ratio of maltose to maltulose and furosine as quality parameters for infant formula

Nonenzymic browning reactions in commercial infant formulas were evaluated through their furosine content as well as the isomeric disaccharides formed during processing. Lactulose was observed only in samples containing appreciable amounts of lactose, whereas maltulose was present in all samples due to the isomerization of maltose. Because formation of maltulose depends on the initial amount of maltose present, the ratio maltose/maltulose was used for comparative purposes. The ratio maltose/maltulose varied within a wide range, 27-167; therefore, low values in maltose/maltulose ratio may indicate severe processing conditions during manufacture, whereas high values may indicate mild processing conditions. Variable amounts of furosine content in samples with similar maltose/maltulose ratios may be attributed to different conditions used during storage. Levels of furosine higher than those reported for milk powder were detected in most studied samples. Determination of both furosine and maltose/maltulose ratio would yield information retrospectively about the heat treatment applied during processing and the storage conditions of commercial infant formula.     

A comparative study of the precipitation acidity in two cities in Cantabria (Spain) with different degrees of industrialization

A study was carried out on the pH values obtained from bulk (wet and dry) precipitation samples collected simultaneously in two cities in Cantabria (Spain), Santander and Torrelavega over a period of a year. It was observed that the annual weighted average values for the pH of precipitation, 4.42 and 4.21, respectively, differ significantly although the difference is less than that existing between their pollution levels. The difference is greater when total annual H⁺ depositions are compared. The study also demonstrated the strong influence of long-range transport effect on acidity and to a lesser degree the effect of mid-range transport.     

2-Furoylmethyl amino acids and hydroxymethylfurfural as indicators of honey quality

Determination of changes in 2-furoylmethyl amino acids and hydroxymethylfurfural during the storage of four honey samples at 25 and 35 degrees C during 12 months was achieved to assess the potential use of both parameters, singly or in combination, as quality indicators. 2-Furoylmethyl amino acids increased during storage at both temperatures, whereas hydroxymethylfurfural only presented slight variations during storage at 25 degrees C but increased noticeably at 35 degrees C. The study of 2-furoylmethyl amino acids in 49 commercial honeys revealed that 2-furoylmethyl lysine (furosine) was present in all samples, whereas 2-furoylmethyl derivatives of arginine, GABA, and proline were only present in seven samples. Hydroxymethylfurfural can be considered as a good indicator of heat treatments applied to honey samples, whereas 2-furoylmethyl amino acids can be used as suitable markers of the storage period. The use of both parameters can be useful to detect adulteration with invert syrups, excessive heat treatments, or prolonged storage of honey samples.     

Differentiation of the crystalline lens in explants of the chicken embryo in the absence of the hypoblast and optical vesicle

In vitro studies of lens formation in chick embryo have suggested the action of two factors leading the lens induction in the cephalic ectoderm in the absence of optic vesicle: preliminary instructive specific stimulus (homotypic endo-mesoderm) and permissive unspecific stimulus (heterotypic mesenchymes). In order to detect the true capacities of tissues that exert this influence in the cultural condition, series of in vitro experiments were planned. Exclusion experiments: explants including presumptive lens ectoderm were cultured previous to a progressive exclusion of adjacent tissues to trigger lens formation (endoderm, mesoderm and neural tissue), from stage 1 to 7 of HAMBURGER/HAMILTON. Recombinant experiments: Recombinations of caudal epiblast with cephalic hypoblast from blastoderms stages 3, 4 and 5; and recombinations of cardiac mesoderm stage 7 with trunk ectoderm stage 11, were cultured in close association. Lens and lentoids were formed in the presumptive lens ectoderm, even when endoderm, neural tissue and optic vesicle were excluded, but always in presence of subjacent mesoderm. Observation of cephalic epiblast after to be separated mechanically from the underlying tissues showed that the presumptive cardiac mesoderm remains in contact with the epiblast. Beside the cardiac area was capable of forming lens bodies in contact with the trunk ectoderm. It was concluded that the cardiac mesoderm is able to exert a instructive specific stimulus and a permissive unspecific stimulus during in vitro lens formation.