Socio-Spatial Relationships in Dairy Cows

Farm animals may serve as models for evaluating social networks in a controlled environment. We used an automated system to track, at fine temporal and spatial resolution (once per minute, +/- 50 cm) every individual in six herds of dairy cows (Bos taurus). We then analysed the data using social network analyses. Relationships were based on non-random attachment and avoidance relationships in respect to synchronous use and distances observed in three different functional areas (activity, feeding and lying). We found that neither synchrony nor distance between cows was strongly predictable among the three functional areas. The emerging social networks were tightly knit for attachment relationships and less dense for avoidance relationships. These networks loosened up from the feeding and lying area to the activity area, and were less dense for relationships based on synchronicity than on median distance with respect to node degree, relative size of the largest cluster, density and diameter of the network. In addition, synchronicity was higher in dyads of dairy cows that had grown up together and shared their last dry period. This last effect disappeared with increasing herd size. Dairy herds can be characterized by one strongly clustered network including most of the herd members with many non-random attachment and avoidance relationships. Closely synchronous dyads were composed of cows with more intense previous contact. The automatic tracking of a large number of individuals proved promising in acquiring the data necessary for tackling social network analyses.

Comparative expression profiling of E. coli and S. aureus inoculated primary mammary gland cells sampled from cows with different genetic predispositions for somatic cell score

BACKGROUND: During the past ten years many quantitative trait loci (QTL) affecting mastitis incidence and mastitis related traits like somatic cell score (SCS) were identified in cattle. However, little is known about the molecular architecture of QTL affecting mastitis susceptibility and the underlying physiological mechanisms and genes causing mastitis susceptibility. Here, a genome-wide expression analysis was conducted to analyze molecular mechanisms of mastitis susceptibility that are affected by a specific QTL for SCS on Bos taurus autosome 18 (BTA18). Thereby, some first insights were sought into the genetically determined mechanisms of mammary gland epithelial cells influencing the course of infection. METHODS: Primary bovine mammary gland epithelial cells (pbMEC) were sampled from the udder parenchyma of cows selected for high and low mastitis susceptibility by applying a marker-assisted selection strategy considering QTL and molecular marker information of a confirmed QTL for SCS in the telomeric region of BTA18. The cells were cultured and subsequently inoculated with heat-inactivated mastitis pathogens Escherichia coli and Staphylococcus aureus, respectively. After 1, 6 and 24 h, the cells were harvested and analyzed using the microarray expression chip technology to identify differences in mRNA expression profiles attributed to genetic predisposition, inoculation and cell culture. RESULTS: Comparative analysis of co-expression profiles clearly showed a faster and stronger response after pathogen challenge in pbMEC from less susceptible animals that inherited the favorable QTL allele 'Q' than in pbMEC from more susceptible animals that inherited the unfavorable QTL allele 'q'. Furthermore, the results highlighted RELB as a functional and positional candidate gene and related non-canonical Nf-kappaB signaling as a functional mechanism affected by the QTL. However, in both groups, inoculation resulted in up-regulation of genes associated with the Ingenuity pathways 'dendritic cell maturation' and 'acute phase response signaling', whereas cell culture affected biological processes involved in 'cellular development'. CONCLUSIONS: The results indicate that the complex expression profiling of pathogen challenged pbMEC sampled from cows inheriting alternative QTL alleles is suitable to study genetically determined molecular mechanisms of mastitis susceptibility in mammary epithelial cells in vitro and to highlight the most likely functional pathways and candidate genes underlying the QTL effect.

Dual origins of dairy cattle farming - evidence from a comprehensive survey of European Y-chromosomal variation

BACKGROUND: Diversity patterns of livestock species are informative to the history of agriculture and indicate uniqueness of breeds as relevant for conservation. So far, most studies on cattle have focused on mitochondrial and autosomal DNA variation. Previous studies of Y-chromosomal variation, with limited breed panels, identified two Bos taurus (taurine) haplogroups (Y1 and Y2; both composed of several haplotypes) and one Bos indicus (indicine/zebu) haplogroup (Y3), as well as a strong phylogeographic structuring of paternal lineages. METHODOLOGY AND PRINCIPAL FINDINGS: Haplogroup data were collected for 2087 animals from 138 breeds. For 111 breeds, these were resolved further by genotyping microsatellites INRA189 (10 alleles) and BM861 (2 alleles). European cattle carry exclusively taurine haplotypes, with the zebu Y-chromosomes having appreciable frequencies in Southwest Asian populations. Y1 is predominant in northern and north-western Europe, but is also observed in several Iberian breeds, as well as in Southwest Asia. A single Y1 haplotype is predominant in north-central Europe and a single Y2 haplotype in central Europe. In contrast, we found both Y1 and Y2 haplotypes in Britain, the Nordic region and Russia, with the highest Y-chromosomal diversity seen in the Iberian Peninsula. CONCLUSIONS: We propose that the homogeneous Y1 and Y2 regions reflect founder effects associated with the development and expansion of two groups of dairy cattle, the pied or red breeds from the North Sea and Baltic coasts and the spotted, yellow or brown breeds from Switzerland, respectively. The present Y1-Y2 contrast in central Europe coincides with historic, linguistic, religious and cultural boundaries.

Analysis of key molecules of the innate immune system in mammary epithelial cells isolated from marker-assisted and conventionally selected cattle

Mastitis is the most prevalent infectious disease in dairy herds. Breeding programs considering mastitis susceptibility were adopted as approaches to improve udder health status. In recent decades, conventional selection criteria based on phenotypic characteristics such as somatic cell score in milk have been widely used to select animals. Recently, approaches to incorporate molecular information have become feasible because of the detection of quantitative trait loci (QTL) affecting mastitis resistance. The aims of the study were to explore molecular mechanisms underlying mastitis resistance and the genetic mechanisms underlying a QTL on Bos taurus chromosome 18 found to influence udder health. Primary cell cultures of mammary epithelial cells from heifers that were selected for high or low susceptibility to mastitis were established. Selection based on estimated pedigree breeding value or on the basis of marker-assisted selection using QTL information was implemented. The mRNA expression of 10 key molecules of the innate immune system was measured using quantitative real-time PCR after 1, 6, and 24 h of challenge with heat-inactivated mastitis pathogens (Escherichia coli and Staphylococcus aureus) and expression levels in the high and low susceptibility groups were compared according to selection criteria. In the marker-assisted selection groups, mRNA expression in cells isolated from less-susceptible animals was significantly elevated for toll-like receptor 2, tumor necrosis factor-alpha, IL-1beta, IL-6, IL-8, RANTES (regulated upon activation, normal t-cell expressed and secreted), complement factor C3, and lactoferrin. In the estimated pedigree breeding value groups, mRNA expression was significantly elevated only for V-rel reticuloendotheliosis viral oncogene homolog A, IL-1 beta, and RANTES. These observations provide first insights into genetically determined divergent reactions to pathogens in the bovine mammary gland and indicate that the application of QTL information could be a successful tool for the selection of animals resistant to mastitis.

Mitochondrial diversity and the origins of African and European cattle.

The nature of domestic cattle origins in Africa are unclear as archaeological data are relatively sparse. The earliest domesticates were humpless, or Bos taurus, in morphology and may have shared a common origin with the ancestors of European cattle in the Near East. Alternatively, local strains of the wild ox, the aurochs, may have been adopted by peoples in either continent either before or after cultural influence from the Levant. This study examines mitochondrial DNA displacement loop sequence variation in 90 extant bovines drawn from Africa, Europe, and India. Phylogeny estimation and analysis of molecular variance verify that sequences cluster significantly into continental groups. The Indian Bos indicus samples are most markedly distinct from the others, which is indicative of a B. taurus nature for both European and African ancestors. When a calibration of sequence divergence is performed using comparisons with bison sequences and an estimate of 1 Myr since the Bison/Bos Leptobos common ancestor, estimates of 117-275,000 B.P. and 22-26,000 B.P. are obtained for the separation between Indians and others and between African and European ancestors, respectively. As cattle domestication is thought to have occurred approximately 10,000 B.P., these estimates suggest the domestication of genetically discrete aurochsen strains as the origins of each continental population. Additionally, patterns of variation that are indicative of population expansions (probably associated with the domestication process) are discernible in Africa and Europe. Notably, the genetic signatures of these expansions are clearly younger than the corresponding signature of African/European divergence.

Feeding strategies for grain-fed cattle in a hot environment

Six Bos taurus (Hereford) steers (body weight 324 22 kg) were used in a 45-day study with a replicated 3 x 3 Latin-square design. Three treatments [ad libitum feeding (ADLIB); limit feeding, 85% of ad libitum (LIMIT); bunk management feeding where steers were only given access to feed from 1600 to 0800 hours the following day (BUNK)] were imposed over 3 periods, with 2 steers assigned to each treatment in each period. Cattle were managed in a temperature-controlled metabolism unit and were exposed to both thermoneutral (17.7degreesC-26.1degreesC) and hot (16.7degreesC-32.9degreesC) environmental conditions. By design, during the thermoneutral period, the ADLIB cattle displayed greater intake (P < 0.05) than the LIMIT group, with the BUNK group being intermediate. However, during the hot period, both the LIMIT and BUNK treatment groups increased feed intake 4-5%, whereas feed intake of the ADLIB treatment group declined nearly 2%. During both periods respiration rate (RR, breath/min) followed the same pattern that was observed for feed intake, with the greatest (P < 0.05) RR found in the ADLIB treatment group (81.09 and 109.55, thermoneutral and hot, respectively) and lowest (P < 0.05) RR in the LIMIT treatment group (74.47 and 102.76, thermoneutral and hot, respectively). Rectal temperature (RT) did not differ among treatments during the thermoneutral period or the first hot day, although during the thermoneutral period the ADLIB treatment group did tend to display a lower RT, possibly as a result of other physiological processes (pulse rate and RR) aiding to keep RT lower. During the hot period, differences in RT were found on Day 5, with the LIMIT cattle having lower (P < 0.10) RT (38.92degreesC) than the ADLIB (39.18degreesC) cattle, with BUNK cattle RT (39.14degreesC) being intermediate. However, when hourly data were examined, the ADLIB cattle had greater(P < 0.05) RT than the BUNK and LIMIT at 1800 hours and greater RT (P < 0.05) than the LIMIT group at 1400, 1500, and 1600 hours. Clearly, a change in diurnal RT pattern was obtained by using the LIMIT and BUNK feeding regimen. Both of these groups displayed a peak RT during the hot conditions, between 2100 and 2200 hours, whereas the ADLIB group displayed a peak RT between 1400 and 1500 hours, a time very close to when peak climatic stress occurs. Based on these results it is apparent that feedlot managers could alleviate the effects of adverse hot weather on cattle by utilising either a limit-feeding regimen or altering bunk management practices to prevent feed from being consumed several hours prior to the hottest portion of the day.

Prevalence, severity, and heritability of Stephanofilaria lesions on cattle in central and southern Queensland, Australia

Observations of cattle in central and southern Queensland are collated to de. ne the prevalence and area of Stephanofilaria lesions associated with infestations of the buffalo fly, Haematobia irritans exigua. The observations were made on herds that were being used for other purposes. In a survey of similar to 1500 animals at Belmont in central Queensland in 1982, 98% of cows and 70% of calves had lesions. Most lesions were on the neck and dewlap and 10% were raw and weeping at the time of sampling. The total area of lesions per animal was strongly related to cattle breed and age. Old Bos taurus animals had the greatest area of lesions, whereas young Bos indicus had the least. Heritability estimates were low, averaging 0.01 for calves and 0.18 for cows. A smaller survey of cows and steers at Craighoyle in central Queensland in 1986 showed a higher numbers of lesions and positive correlations between the total lesion area and animal size. The lesion area increased with tick survival, suggesting that tick-resistant animals are also resistant to Stephanofilaria infection. Steers had smaller areas of lesions than cows, as found previously with cattle ticks. Long-term monitoring observations in central and southern Queensland between 1981 and 1986 showed that the total area of lesions was seasonal with a peak in late summer, consistent with the seasonal incidence of buffalo fly. Animals segregated into Low and High lesion herds maintained their differences over time. The lesions penetrated the dermis of the cattle hides and rendered the affected area unusable, but few lesions occurred on valuable parts of the hide so such economic effects are likely to be insignificant. One animal nearly died of a haemorrhage from a lesion on the dewlap and had to be treated. The results can inform policy on buffalo fly control, and biosecurity preparations in relation to the potential establishment of the OldWorld screw-worm fly, Chrysomyia bezziana, in Australia, which will be facilitated by the lesions. The results emphasise the significant animal welfare and biosecurity risks posed by the lesions in northern Australia.

La geochimica degli isotopi stabili applicata a reperti scheletrici fossili per ricostruzioni paleoambientali (sito archeologico di Arslantepe –Turchia).

Il sito archeologico di Arslantepe (provincia di Malatya, Turchia) rappresenta un caso di studio di potenziale interesse per l’interazione tra i mutamenti climatici e la storia della civiltà. Il sito, occupato quasi ininterrottamente per un periodo di tempo relativamente lungo (6250-2700 BP), ha fornito una grande quantità di reperti ossei, distribuiti lungo una stratigrafia archeologica relativamente dettagliata e supportata da datazioni al radiocarbonio. Tali reperti, indagati con le tecniche della geochimica degli isotopi stabili, possono costituire degli efficaci proxy paleoclimatici. In questo lavoro è stata studiata la composizione isotopica di 507 campioni di resti ossei umani e animali (prevalentemente pecore, capre, buoi). I rapporti isotopici studiati sono relativi a ossigeno (δ18Ocarb, δ18Oph), carbonio (δ13Ccarb, δ13Ccoll) e azoto (δ15N), misurati nella frazione minerale e organica dell’osso; la variabilità nel tempo di questi parametri, principalmente legati alla paleonutrizione, può essere correlata, direttamente o indirettamente, a cambiamenti dei parametri ambientali quali temperatura e umidità atmosferiche. I risultati indicano che la dieta degli animali selvatici e domestici di Arslantepe era quasi esclusivamente a base di piante a ciclo fotosintetico C3, generalmente tipiche di climi umidi o temperati. La presenza di piante C4, più tipiche di condizioni aride, sembrerebbe essere riconoscibile solamente nella dieta del bue (Bos taurus). La dieta umana era esclusivamente terrestre a base di cereali e carne di caprini con una percentuale esigua o del tutto assente di carne di maiale e bue. Dal punto di vista paleoclimatico il principale risultato del lavoro consiste nel riconoscimento della preservazione di un segnale paleoclimatico a lungo termine (δ18OW, composizione isotopica dell’ossigeno dell’acqua ingerita), che identifica un massimo relativo di umidità attorno al 5000 BP e che si correla, per andamento e ampiezza della variazione a record paleoclimatici di sedimenti lacustri collocati in regioni adiacenti all’area di studio. Sulla base del confronto dei tre segnali isotopici sono state inoltre riconosciute due anomalie climatiche siccitose a breve termine, apparentemente riferibili a due episodi di aridità a scala regionale documentati in letteratura.

Aplicación de técnicas de PCR en tiempo real a la trazabilidad y autenticidad de los piensos

La trazabilidad y el correcto etiquetado de los piensos y sus ingredientes son factores esenciales para prevenir fraudes y garantizar la seguridad alimentaria. En el ámbito de la lucha contra las Encefalopatías Espongiformes Transmisibles (EETs), la prohibición de la Unión Europea (UE) de alimentar a rumiantes y otros animales de granja con harinas de carne y huesos derivadas de animales, hace necesaria la disponibilidad de metodologías que permitan identificar el origen de las materias primas e ingredientes presentes en los piensos. El método oficial de análisis microscópico tradicionalmente empleado para este fin presenta limitaciones a la hora de diferenciar entre los huesos de mamíferos y de aves, así como para determinar el origen animal específico de las partículas detectadas. Por ello, una de las prioridades de la UE en los últimos años ha sido potenciar la búsqueda y desarrollo de técnicas analíticas alternativas que permitan la detección específica de todos los componentes que integran los piensos. Teniendo en cuenta estos aspectos, en esta Tesis Doctoral se han desarrollado técnicas de PCR en tiempo real con sondas TaqMan® para el control de autenticidad y trazabilidad de ingredientes de origen animal utilizados en la fabricación de los piensos. Las especies objeto de este trabajo han sido: vaca (Bos taurus), oveja (Ovis aries), cabra (Capra hircos), grupo rumiante, cerdo (Sus scrofa), pollo (Gallus gallos), pavo (Meleagris g-allopavo), pato (Anal platyrhynchos x Cairina moschata), oca (Anser anser), grupo aviar, caballo (Equus caballus), conejo (Oryctolagus cuniculus), liebre (Lepus capensis), grupo lepórido (conejo y liebre) y pescados...

Avaliação da viabilidade da aplicação de técnicas biomoleculares na autenticação de produtos alimentares

A carne continua a ser a fonte proteica mais comum no quotidiano das pessoas. Além disso, os produtos cárneos processados apresentam-se como uma mais-valia nas suas vidas agitadas. Este tipo de produto torna difícil a diferenciação das carnes utilizadas na sua confecção, sendo por isso propícios a adulteração. A Reacção em Cadeia da Polimerase (PCR) tem ganho cada vez mais importância nos laboratórios de biologia molecular, revelando-se uma técnica de análise rápida, sensível e altamente específica na identificação de espécies em produtos alimentares. No entanto, vários factores podem interferir com o processo de amplificação, pelo que alguns cuidados devem ser implementados desde a aquisição da amostra a analisar, ao seu acondicionamento e posterior extração de ADN. Existem inúmeros protocolos de extração de ADN, devendo para cada estudo avaliar-se e optar-se pelo mais adequado, considerando a finalidade estabelecida para a amostra extraída. O trabalho laboratorial apresentado nesta dissertação baseou-se em três etapas principais. Inicialmente, avaliaram-se diferentes protocolos de extração de ADN, utilizando-se amostras de carne adquiridas num talho. Entre os protocolos testados, o método de Brometo de Cetil-Trimetil-Amónio (CTAB) modificado foi o que permitiu obter amostras de ADN com maior concentração e elevado nível de pureza. Posteriormente, foram testados e optimizados diferentes protocolos de amplificação, por PCR em tempo real, para a detecção das espécies Bos taurus (vaca), Sus scrofa (porco), Equus caballus (cavalo) e Ovis aries (ovelha). Foram empregues primers específicos de espécie para a detecção de genes mitocondriais e genómicos, consoante cada protocolo. Para o caso concreto do porco, foi efectuada a avaliação de dois protocolos, singleplex com EvaGreen® e tetraplex com AllHorse, para possível aplicação dos mesmos na sua quantificação. Os resultados demonstraram elevada especificidade e sensibilidade das reacções para esta espécie, permitindo a sua detecção até um limite de 0,001 ng e 0,1%, respectivamente. Somente a primeira metodologia se mostrou adequada para quantificação. Por último, as metodologias sugeridas foram aplicadas com sucesso na análise de 4 amostras comerciais de hambúrgueres, tendo-se verificado a consistência da rotulagem em todos os casos, no que concerne a composição em termos de espécies animais. O interesse de trabalhos neste âmbito recai na importância da autenticidade dos rótulos de produtos alimentares, principalmente nos produtos cárneos, para segurança dos consumidores e salvaguarda dos produtores.

Adição de Ácido Linoleico conjugado (CLA) no diluidor de congelação de sémen de touro, recolhido Post-Mortem, das raças Holstein Frísia, Aberdeen Angus, Charolês e Limousine

Dissertação de Mestrado, Engenharia Zootécnica, 7 de Abril de 2016, Universidade dos Açores.

Relação entre caracteres morfológicos e produtivos em vacas Holstein-Frísia em S. Miguel

Dissertação de Mestrado, Engenharia Zootécnica, 18 de Julho de 2016, Universidade dos Açores.