Aplicação da microtomografia de raios-x laboratorial e síncrotron em estudos in vitro utilizando dente bovino e ciclagem de pH

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

"Cara inchada" of cattle, an infectious, apparently soil antibiotics-dependant periodontitis in Brazil

The objective of this review on the investigation of "cara inchada" in cattle (CI), pursued over the last 30 years, was to elucidate the pathogenicity of the disease and come to proper conclusions on its etiology. CI has been widely considered to be of nutritional origin, caused primarily by mineral deficiency or imbalance. However, the disease consists of a rapidly progressive periodontitis, affecting the periodontal tissues at the level of the premolars and molars during the period of tooth eruption generally starting in young calves. The disease led to great economic losses for farmers in central-western Brazil, after the occupation of new land for cattle raising in the 1960s and 1970s. The lateral enlargement of the maxillary bones of affected calves gave the disease the popular name of "cara inchada", i.e., swollen or enlarged face. The enlargement was found to be due to a chronic ossifying periostitis resulting from the purulent alveolitis of CI. Black-pigmented non-saccharolytic Bacteroides melaninogenicus, always together with Actinomyces (Corynebacterium) pyogenes, were isolated in large numbers from the periodontal lesions. B. melaninogenicus could be isolated in small numbers also from the marginal gingiva of a few healthy calves maintained on CI-free farms. "In vitro"-assays showed that streptomycin and actinomycin, as well as the supernatants of cultivates of actinomycetes from soils of CI-prone farms, applied in subinhibitory concentrations to the bacteria tested, enhanced significantly (up to 10 times) the adherence of the black-pigmented B.melaninogenicus to epithelial cells of the bovine gingiva. The antibiotics are apparently produced in large quantities by the increased number of soil actinomycetes, including the genus Streptomyces, that develop when soil microflora are modified by cultivating virgin forest or "Cerrado" (tree-savanna) for the first time for cattle grazing. The epidemiology of CI now provides strong evidence that the ingestion with the forage of such antibiotics could possibly be an important determinant factor for the onset and development of this infectious periodontitis. The antibiotic enhanced adherence of B.melaninogenicus to the sulcus-epithelium of the marginal gingiva, is thought to allow it to colonize, form a plaque and become pathogenic. There is experimental evidence that this determinant factor for the development of the periodontitis is present also in the milk of the mothers of CI-diseased calves. It has been shown that the bacteria isolated from the periodontal CI-lesions produce enzymes and endotoxins capable of destroying the periodontal tissues. The epidemiology of CI, with its decline in incidence and its disappearance after several years, could be explained by the fact that the former equilibrium of the microflora of the once undisturbed virgin soil has been reached again and that the number of antibiotic producing actinomycetes has been anew reduced. By this reasoning and all the data available, CI should be considered as a multifactorial infectious disease, caused primarily by the anaerobic black-pigmented non-saccharolytic Bacteroides melaninogenicus, always together with the micro-anaerobic Actinomyces pyogenes. Accordingly, the onset and development of the infectious periodontitis is apparently determined by ingestion with the forage of subinhibitory concentrations of antibiotics produced in recently cultivated virgin soils. This hypothesis is supported by the recent observation of renewed outbreaks of CI-periodontitis in former CI-prone areas, following fresh cultivation after many years. The infectious nature of CI is confirmed by trials in which virginiamycin was used efficiently for the oral treatment of CI-diseased cattle. Previously it has been shown, that spiramycin and virginiamycin, used as additives in mineral supplements, prevented CI-periodontitis.

Season of birth and sheep husbandry in late Roman and Medieval coastal Flanders: a pilot study using tooth enamel d18O analysis

From the early Roman period, there is archaeological evidence for the exploitation of the Flemish coastal plain (Belgium) for a range of activities, such as sheep herding on the then developing salt-marshes and salt-meadows for the production of wool. During the early Middle Ages, this culminated in the establishment of dedicated ‘sheep estates’. This phase of exploitation was followed by extensive drainage and land reclamation measures in the high Medieval period, transforming areas into grassland, suited for cattle breeding. As part of a larger project investigating the onset, intensification and final decline of sheep management in coastal Flanders in the historical period, this pilot study presents the results of sequential sampling and oxygen isotope analysis of a number of sheep teeth (M2, n = 8) from four late Roman and Medieval sites (dating from 4th to 15th century AD), in order to assess potential variations in season of birth between the different sites and through time. In comparison with published data from herds of known birth season, incremental enamel data from the Flemish sites are consistent with late winter/spring births, with the possibility of some instances of slightly earlier parturition. These findings suggest that manipulation of season of birth was not a feature of the sheep husbandry-based economies of early historic Flanders, further evidencing that wool production was the main purpose of contemporary sheep rearing in the region. Manipulation of season of birth is not likely to have afforded economic advantage in wool-centred economies, unlike in some milk- or meat-based regimes.

Cara inchada of cattle, an infectious, apparently soil antibiotics-dependant periodontitis in Brazil

O objetivo desta revisão das pesquisas sobre a cara inchada dos bovinos (CI), realizadas no decorrer dos últimos 30 anos, é de elucidar melhor a sua etiologia. A CI geralmente tem sido considerada de origem nutricional, causada primariamente por deficiência ou desequilíbrio mineral. A doença caracteriza-se por uma periodontite rapidamente progressiva, que afeta os tecidos peridentários a nível dos premolares e molares no período de erupção dos dentes e que se inicia geralmente em bezerros jovens. A doença causou grandes perdas econômicas aos pecuaristas da Região Centro-Oeste do Brasil, nas décadas de 1960 e 1970, com a ocupação de novas terras para criação de gado. O freqüente abaulamento lateral dos ossos maxilares nos bezerros, que deu à doença o nome popular de cara inchada, foi demonstrado ser conseqüente à periostite crônica ossificante resultante da alveolite purulenta da CI. Das lesões peridentárias foi isolado, em grande número, Bacteroides melaninogenicus, sempre junto com Actinomyces (Corynebacterium) pyogenes. Bactérias classificadas como pertencentes ao grupo sacarolí-tico e não-sacarolítico dos pigmentados de negro Bacteroides melaninogenicus e Bacteroides spp também foram isoladas, em pequeno número, de bovinos jovens sadios de fazendas CI-negativas. Ensaios in vitro mostraram que os antibióticos estreptomicina e actinomicina, bem como os sobrenadantes de cultivos de actinomicetos do solo de fazendas CI-positivas, aplicadas nas bactérias ensaiadas em concentrações subinibidoras, aumentaram significativamente (até 10 vezes) a aderência de B.melaninogenicus a células epiteliais da gengiva bovina. Esses antibióticos são produzidos no solo em conseqüência de um aumento do número de actinomicetos, incluindo os do gênero Streptomyces, quando há modificação de sua microbiota em áreas previamente ocupadas por mata virgem ou vegetação natural de Cerrado, que foram cultivadas pela primeira vez na formação de pastagem para o gado. em face da epidemiologia da CI, há fortes evidências de que a ingestão desses antibióticos pelos bovinos, junto com a forrageira, seja importante fator desencadeante para o desenvolvimento da periodontite. Através do aumento da aderência de B. melaninogenicus ao epitélio da gengiva marginal, em face da ingestão dos antibióticos pelos animais, as bactérias conseguem colonizar, formar a placa bacteriana e tornar-se patogênicas. Há evidência de que o fator desencadeante (aparentemente, os antibióticos) esteja também presente no leite de vacas-mães de bezerros afetados pela CI. Foi demonstrado que as bactérias envolvidas na periodontite produzem enzimas e endotoxinas capazes de ação destrutiva sobre os tecidos peridentários. A epidemiologia da CI, com a diminuição de sua incidência e o seu desaparecimento no decorrer dos anos, pode ser explicada pelo fato de que o prévio equílibrio da microbiota no solo virgem foi alcançado novamente e a produção dos antibióticos se reduziu. Desta maneira, a CI deve ser considerada como uma periodontite infecciosa multifatorial, causada sobretudo por bactérias anaeróbias pertencentes ao grupo Bacteroides melaninogenicus e, ao que tudo indica, desencadeada pela ingestão contínua, com a forrageira, de concentrações subinibidoras de antibióticos de solos recentememente cultivados. Esta hipótese é reforçada pela observação recente de novos surtos de CI, em áreas anteriormente positivas para a doença, em conseqüência da reforma de pastagens e capineiras após muitos anos. A natureza infecciosa da CI-periodontite foi confirmada através de experimento, em que virginiamicina mostrou-se eficaz no tratamento oral de bovinos afetados pela doença. Os antibióticos espiramicina e virginiamicina, usados como aditivos em suplementos minerais no campo, mostraram-se eficientes na prevenção da CI.

Efficacy of mouth rinses and toothpaste on tooth whitening.

People increasingly desire tooth whitening. Considering the wide range of whitening products on the market, this study evaluated the efficacy of whitening toothpastes and mouth rinses compared with the 10% carbamide peroxide (CP) whitening gel. We obtained 120 cylindrical specimens from bovine teeth, which were darkened for 24 hours in a coffee solution. The color measurement was performed by a spectrophotometer using the CIE L*a*b* system, and specimens were divided into six groups according to the use of the following agents: group 1, conventional fluoridated toothpaste; group 2, Close Up White Now; group 3, Listerine Whitening; group 4, Colgate Plax Whitening; group 5, experimental mouth rinse with Plasdone; and group 6, 10% CP Whiteness Perfect. After the simulation of 12 weeks of treatment for groups 1 to 5 and 14 days of treatment for group 6, the specimens were subjected to a new color reading. Data were subjected to one-way analysis of variance (α=0.05), which showed significant differences among groups after 12 weeks for ΔE (p=0.001). Results of the Tukey test revealed that groups 3, 4, and 6 presented significantly higher color alteration than groups 1, 2, and 5. The whitening toothpaste Close Up White Now and the experimental mouth rinse with Plasdone showed similar color alteration as conventional toothpaste after a 12-week treatment simulation. These groups presented significantly lower color alteration compared with whitening mouth rinses Listerine and Colgate Plax Whitening, which showed similar results to those observed after 14 days of bleaching with 10% CP treatment.

Bovine Teeth in Age Assessment, from Medieval Cattle to Belgian Blue : Methodology, Possibilities and Limitations

The overall aim of the present thesis was to develop and characterise an age assessment method based on incremental lines in dental cementum using contemporary bovine teeth and teeth from archaeological faunal assemblages. The investigations also included two other age assessment methods: tooth wear pattern and macroscopic dental measurements. The first permanent mandibular molar and lower jaws from 70 contemporary cattle of known age and 170 archaeological molar sets from ten different Swedish archaeological sites were used. The following conclusions were drawn: • The number of incremental lines in the dental cementum varied between different parts of the tooth root as well as within one and the same individual. The results from contemporary cattle of known age showed a strong relationship between age and incremental lines in the cementum of the distal part of the mesial root (R2=65.5%) and the known ages of the animals. • With the “best” model variation in age could be explained to 65.5% (R2) by the number of incremental lines. Thus, the remaining age variation (approximately 35%) could not be explained by these lines. Other factors than must thus be responsible. However, with the exception of calves born the present material did not reveal any such significant relationship. • The results from cattle of known age indicate that the method of assessing age on the basis of cemental incremental lines is more reliable than other methods such as tooth wear or tooth measurements. However, by combining counting incremental lines and one variable assessing tooth dimension (tooth height) a slightly stronger relationship could be obtained (R2=74.5%). The results from age assessment of the medieval and post-Reformation cattle emphasize the importance of supplementing any age estimation of archaeological assemblages based on dental indicators with characteristics for the particular assessment model. Furthermore, conclusions based on age assessment with such models can not be drawn with any more detailed time scale than about 2 years leaving at best only 25% (R2) of factors influencing the dental indicator(s) utilized in the model unexplained. The accuracy of the age assessment required by the particular historical context in which the archaeological remains are found should thus decide what level of accuracy should be chosen.

An unusual splice defect in the mitofusin 2 gene (MFN2) is associated with degenerative axonopathy in Tyrolean Grey cattle

Tyrolean Grey cattle represent a local breed with a population size of approximately 5000 registered cows. In 2003, a previously unknown neurological disorder was recognized in Tyrolean Grey cattle. The clinical signs of the disorder are similar to those of bovine progressive degenerative myeloencephalopathy (weaver syndrome) in Brown Swiss cattle but occur much earlier in life. The neuropathological investigation of an affected calf showed axonal degeneration in the central nervous system (CNS) and femoral nerve. The pedigrees of the affected calves suggested a monogenic autosomal recessive inheritance. We localized the responsible mutation to a 1.9 Mb interval on chromosome 16 by genome-wide association and haplotype mapping. The MFN2 gene located in this interval encodes mitofusin 2, a mitochondrial membrane protein. A heritable human axonal neuropathy, Charcot-Marie-Tooth disease-2A2 (CMT2A2), is caused by MFN2 mutations. Therefore, we considered MFN2 a positional and functional candidate gene and performed mutation analysis in affected and control Tyrolean Grey cattle. We did not find any non-synonymous variants. However, we identified a perfectly associated silent SNP in the coding region of exon 20 of the MFN2 gene. This SNP is located within a putative exonic splice enhancer (ESE) and the variant allele leads to partial retention of the entire intron 19 and a premature stop codon in the aberrant MFN2 transcript. Thus we have identified a highly unusual splicing defect, where an exonic single base exchange leads to the retention of the preceding intron. This splicing defect represents a potential explanation for the observed degenerative axonopathy. Marker assisted selection can now be used to eliminate degenerative axonopathy from Tyrolean Grey cattle.

Antimicrobial susceptibility of coagulase-negative Staphylococci isolated from mastitic cattle in Brazil

Um total de 109 cepas de Staphylococci coagulase-negativa foi isolado de leite de vacas com mastite clínica e subclínica, em 35 fazendas, situadas em nove estados brasileiros, no período de fevereiro a maio de 2005. Os isolados foram investigados em relação a susceptibilidade in vitro a diversos agentes antimicrobianos. A resistência à penicilina foi a observação mais freqüente (93,5%), seguida por sulfonamida (88,9%), novobiocina (88,6%) e ampicilina (85,3%). Todas as cepas examinadas mostraram resistência a pelo menos uma das drogas antimicrobianas testadas. Cepas apresentando resistência múltipla foram extremamente comuns, com 10,0% dos microrganismos isolados apresentando resistência a todas as drogas antimicrobianas. Os resultados obtidos indicaram que as cepas de Staphylococci coagulase-negativas, isoladas no Brasil, apresentaram um alto grau de resistência a antimicrobianos. Estes resultados são, provavelmente, uma conseqüência da pressão devida ao uso intensivo de drogas antimicrobianas.

Control of erosive tooth wear: possibilities and rationale

Dental erosion is a type of wear caused by non bacterial acids or chelation. There is evidence of a significant increase in the prevalence of dental wear in the deciduous and permanent teeth as a consequence of the frequent intake of acidic foods and drinks, or due to gastric acid which may reach the oral cavity following reflux or vomiting episodes. The presence of acids is a prerequisite for dental erosion, but the erosive wear is complex and depends on the interaction of biological, chemical and behavioral factors. Even though erosion may be defined or described as an isolated process, in clinical situations other wear phenomena are expected to occur concomitantly, such as abrasive wear (which occurs, e.g, due to tooth brushing or mastication). In order to control dental loss due to erosive wear it is crucial to take into account its multifactorial nature, which predisposes some individuals to the condition.

Effects of dietary energy and nitrogen supplements on rumen fermentation and protozoa population in buffalo and zebu cattle

The effects were assessed of two energy sources in concentrate (ground grain corn vs. citrus pulp) and two nitrogen sources (soybean meal vs. urea) on rumen metabolism in four buffaloes and four zebu cattle (Nellore) with rumen cannula and fed in a 4 × 4 Latin square design with feeds containing 60% sugar cane. Energy supplements had no effect on the rumen ammonia concentration in cattle, but ground grain corn promoted higher ammonia level than citrus pulp in buffalo. Urea produced higher ammonia level than soybean meal in both animal species. On average, the buffaloes maintained a lower rumen ammonia concentration (11.7 mg/dL) than the cattle (14.5 mg/dL). Buffaloes had lower production of acetic acid than cattle (58.7 vs. 61.6 mol/100 mol) and higher of propionic acid (27.4 vs. 23.6 mol/100 mol). There was no difference in the butyric acid production between the buffaloes (13.6 mol/100 mol) and cattle (14.8 mol/100 mol) and neither in the total volatile fatty acids concentration (82.5 vs. 83.6 mM, respectively). The energy or nitrogen sources had no effect on rumen protozoa count in either animal species. The zebu cattle had higher rumen protozoa population (8.8 × 10(5)/mL) than the buffaloes (6.1 × 10(5)/mL). The rumen protozoa population differed between the animal species, except for Dasytricha and Charonina. The buffaloes had a lower Entodinium population than the cattle (61.0 vs 84.9%, respectively) and a greater percentage of species belonging to the Diplodiniinae subfamily than the cattle (28.6 vs. 1.4%, respectively). In cattle, ground corn is a better energy source than citrus pulp for use by Entodinium and Diplodiniinae. In the buffaloes, the Entodinium are favored by urea and Diplodiniinae species by soybean meal.

Timing and sequence of primary tooth eruption in children with cleft lip and palate

OBJECTIVE: To determine the timing and sequence of eruption of primary teeth in children with complete bilateral cleft lip and palate. MATERIAL AND METHODS: This cross-sectional study was conducted at the Hospital for Rehabilitation of Craniofacial Anomalies of the University of São Paulo, Bauru, SP, Brazil, with a sample of 395 children (128 girls and 267 boys) aged 0 to 48 months, with complete bilateral cleft lip and palate. RESULTS: Children with complete bilateral clefts presented a higher mean age of eruption of all primary teeth for both arches and both genders, compared to children without clefts. This difference was statistically signifcant for all teeth, except for the maxillary first molar. Mean age of eruption of most teeth was lower for girls compared to boys. The greatest delay was found for the maxillary lateral incisor, which was the eighth tooth of children with clefts of both genders. Analyzing by gender, the maxillary lateral incisor was the eighth tooth to erupt in girls and the last in boys. CONCLUSION: The results suggest an interference of the cleft on the timing and sequence of eruption of primary teeth.

Genome wide scan for quantitative trait loci affecting tick resistance in cattle (Bos taurus x Bos indicus)

Background : In tropical countries, losses caused by bovine tick Rhipicephalus (Boophilus) microplus infestation have a tremendous economic impact on cattle production systems. Genetic variation between Bos taurus and Bos indicus to tick resistance and molecular biology tools might allow for the identification of molecular markers linked to resistance traits that could be used as an auxiliary tool in selection programs. The objective of this work was to identify QTL associated with tick resistance/susceptibility in a bovine F2 population derived from the Gyr (Bos indicus) x Holstein (Bos taurus) cross. Results: Through a whole genome scan with microsatellite markers, we were able to map six genomic regions associated with bovine tick resistance. For most QTL, we have found that depending on the tick evaluation season (dry and rainy) different sets of genes could be involved in the resistance mechanism. We identified dry season specific QTL on BTA 2 and 10, rainy season specific QTL on BTA 5, 11 and 27. We also found a highly significant genome wide QTL for both dry and rainy seasons in the central region of BTA 23. Conclusions: The experimental F2 population derived from Gyr x Holstein cross successfully allowed the identification of six highly significant QTL associated with tick resistance in cattle. QTL located on BTA 23 might be related with the bovine histocompatibility complex. Further investigation of these QTL will help to isolate candidate genes involved with tick resistance in cattle.

Epidermal growth factor in liposomes may enhance osteoclast recruitment during tooth movement in rats

Objective: To evaluate the effects of local administration of epidermal growth factor (EGF) located within liposomes on recruitment of osteoclasts during mechanical force in rats. Materials and Methods: An orthodontic elastic band was inserted between the left upper first and second molars, to move mesially the first molar. Rats were randomly divided into 4 groups (n = 8): EGF (2 ng/mu L) located within liposomes (group 1), liposomes only (group 2), soluble EGF (2 ng/mu L; group 3), or vehicle alone (group 4). The solutions were injected into the region of the root furcation of the left first molar after elastic band insertion. Tooth movement was measured using a plaster model of the maxilla, and the number of osteoclasts recruited at the pressure side of the first molar was histologically evaluated. Results: Intergroup analysis showed that there was no significant difference between group 2 and group 4 (P >.05) and between group 1 and group 3 (P >.05). However, group 1 and group 3 exhibited greater differences in tooth movement than group 2 and group 4 (P <.05). On the other hand, group 1 showed greater tooth movement than groups 2 and 4 with statistical significance (P <.01). The increase in the number of osteoclasts in group 1 was significantly higher than in the other groups (P <.05). Conclusion: Exogenous EGF-liposome administration has an additive effect when compared with soluble EGF on the rate of osteoclast recruitment, producing faster bone resorption and tooth movement.

An assessment of population structure in eight breeds of cattle using a whole genome SNP panel

Background: Analyses of population structure and breed diversity have provided insight into the origin and evolution of cattle. Previously, these studies have used a low density of microsatellite markers, however, with the large number of single nucleotide polymorphism markers that are now available, it is possible to perform genome wide population genetic analyses in cattle. In this study, we used a high-density panel of SNP markers to examine population structure and diversity among eight cattle breeds sampled from Bos indicus and Bos taurus. Results: Two thousand six hundred and forty one single nucleotide polymorphisms ( SNPs) spanning all of the bovine autosomal genome were genotyped in Angus, Brahman, Charolais, Dutch Black and White Dairy, Holstein, Japanese Black, Limousin and Nelore cattle. Population structure was examined using the linkage model in the program STRUCTURE and Fst estimates were used to construct a neighbor-joining tree to represent the phylogenetic relationship among these breeds. Conclusion: The whole-genome SNP panel identified several levels of population substructure in the set of examined cattle breeds. The greatest level of genetic differentiation was detected between the Bos taurus and Bos indicus breeds. When the Bos indicus breeds were excluded from the analysis, genetic differences among beef versus dairy and European versus Asian breeds were detected among the Bos taurus breeds. Exploration of the number of SNP loci required to differentiate between breeds showed that for 100 SNP loci, individuals could only be correctly clustered into breeds 50% of the time, thus a large number of SNP markers are required to replace the 30 microsatellite markers that are currently commonly used in genetic diversity studies.

Multi-trait and random regression mature weight heritability and breeding value estimates in Nelore cattle

Mature weight breeding values were estimated using a multi-trait animal model (MM) and a random regression animal model (RRM). Data consisted of 82 064 weight records from 8 145 animals, recorded from birth to eight years of age. Weights at standard ages were considered in the MM. All models included contemporary groups as fixed effects, and age of dam (linear and quadratic effects) and animal age as covariates. In the RRM, mean trends were modelled through a cubic regression on orthogonal polynomials of animal age and genetic maternal and direct and maternal permanent environmental effects were also included as random. Legendre polynomials of orders 4, 3, 6 and 3 were used for animal and maternal genetic and permanent environmental effects, respectively, considering five classes of residual variances. Mature weight (five years) direct heritability estimates were 0.35 (MM) and 0.38 (RRM). Rank correlation between sires' breeding values estimated by MM and RRM was 0.82. However, selecting the top 2% (12) or 10% (62) of the young sires based on the MM predicted breeding values, respectively 71% and 80% of the same sires would be selected if RRM estimates were used instead. The RRM modelled the changes in the (co) variances with age adequately and larger breeding value accuracies can be expected using this model.