A comparison of the estrous behavior of Holstein-Friesian cows when cubicle-housed and at pasture

This study compared estrous behavior of dairy cows kept in cubicle housing and fed a total mixed ration diet (HOUSED treatment) with that of cows kept at pasture with concentrate supplementation (PASTURE treatment). Behavior was compared both in the 48 h around standing estrus and during the standing estrus period. The 23 spring-calving Holstein-Friesians in each treatment were observed directly three times per day for nine weeks. The occurrence of nine selected behaviors associated with estrus was recorded during 20 min observation sessions. Twelve standing estrus events from each treatment were selected for analysis of the frequency of these nine behaviours over the 48 h around standing estrus. Milk progesterone profiles were used to confirm the dates of standing estrus events. Attempting to mount other cows, sniffing the anogenital region of other cows, resting the chin on other cows, receiving chin rests and head-to-head butts all showed significant changes in frequency in the 48 h around standing estrus in both treatments, reaching a peak during standing estrus (P ≤ 0.05). Mounting other cows increased significantly in the PASTURE treatment around standing estrus (P <0.001), but not in the HOUSED treatment. The frequency of ano-genital sniffs received by the animals in the PASTURE treatment also increased significantly around standing estrus (P <0.01) but not in the HOUSED treatment. When the animals were in standing estrus there was a significantly higher frequency of standing to be mounted in PASTURE than in HOUSED cows (median (q1, q3) PASTURE = 2.5 (1.0, 3.0), HOUSED = 0.0 (0.0, 1.0)) (P <0.01), but no difference in the frequency of the other eight sexual behaviors recorded. HOUSED cows did not exhibit the same increase in mounting during the standing estrus period as PASTURE cows and received fewer mounts in observation sessions during standing estrus. These results have implications for the use of estrus detection systems that rely solely on mounting behavior in cubicle-housed dairy cows. © 2012 Elsevier Inc.

Sensitive and specific detection of viable Mycobacterium avium subsp. paratuberculosis in raw milk by the Peptide-mediated magnetic separation (PMS)-phage assay

This study describes further validation of a previously described Peptide-mediated magnetic separation (PMS)-Phage assay, and its application to test raw cows’ milk for presence of viable Mycobacterium avium subsp. paratuberculosis (MAP). The inclusivity and exclusivity of the PMS-phage assay were initially assessed, before the 50% limit of detection (LOD50) was determined and compared with those of PMS-qPCR (targeting both IS900 and f57) and PMS-culture. These methods were then applied in parallel to test 146 individual milk samples and 22 bulk tank milk samples from Johne’s affected herds. Viable MAP were detected by the PMS-phage assay in 31 (21.2%) of 146 individual milk samples (mean plaque count of 228.1 PFU/50 ml, range 6-948 PFU/50 ml), and 13 (59.1%) of 22 bulk tank milks (mean plaque count of 136.83 PFU/50 ml, range 18-695 PFU/50 ml). In contrast, only 7 (9.1%) of 77 individual milks and 10 (45.4%) of 22 bulk tank milks tested PMS-qPCR positive, and 17 (11.6%) of 146 individual milks and 11 (50%) of 22 bulk tank milks tested PMS-culture positive. The mean 50% limits of detection (LOD50) of the PMS-phage, PMS-IS900 qPCR and PMS-f57 qPCR assays, determined by testing MAP-spiked milk, were 0.93, 135.63 and 297.35 MAP CFU/50 ml milk, respectively. Collectively, these results demonstrate that, in our laboratory, the PMS-phage assay is a sensitive and specific method to quickly detect the presence of viable MAP cells in milk. However, due to its complicated, multi-step nature, the method would not be a suitable MAP screening method for the dairy industry.

Portable “lab-on-chip” platform for bovine mastitis diagnosis in raw milk

Tese de Doutoramento em Ciências Veterinárias, Especialidade de Ciências Biológicas e Biomédicas

Evaluation of physicochemical and glycaemic properties of commercial plant-based milk substitutes

The market for plant-based dairy-type products is growing as consumers replace bovine milk in their diet, for medical reasons or as a lifestyle choice. A screening of 17 different commercial plant-based milk substitutes based on different cereals, nuts and legumes was performed, including the evaluation of physicochemical and glycaemic properties. Half of the analysed samples had low or no protein contents (<0.5 %). Only samples based on soya showed considerable high protein contents, matching the value of cow’s milk (3.7 %). An in-vitro method was used to predict the glycaemic index. In general, the glycaemic index values ranged from 47 for bovine milk to 64 (almond-based) and up to 100 for rice-based samples. Most of the plant-based milk substitutes were highly unstable with separation rates up to 54.39 %/h. This study demonstrated that nutritional and physicochemical properties of plant-based milk substitutes are strongly dependent on the plant source, processing and fortification. Most products showed low nutritional qualities. Therefore, consumer awareness is important when plant-based milk substitutes are used as an alternative to cow’s milk in the diet.

A Survey of the Relationship between Climatic Heat Stress Indices and Fundamental Milk Components Considering Uncertainty

The main purpose of this study is to assess the relationship between four bioclimatic indices for cattle (environmental stress, heat load, modified heat load, and respiratory rate predictor indices) and three main milk components (fat, protein, and milk yield) considering uncertainty. The climate parameters used to calculate the climate indices were taken from the NASA-Modern Era Retrospective-Analysis for Research and Applications (NASA-MERRA) reanalysis from 2002 to 2010. Cow milk data were considered for the same period from April to September when the cows use the natural pasture. The study is based on a linear regression analysis using correlations as a summarizing diagnostic. Bootstrapping is used to represent uncertainty information in the confidence intervals. The main results identify an interesting relationship between the milk compounds and climate indices under all climate conditions. During spring, there are reasonably high correlations between the fat and protein concentrations vs. the climate indices, whereas there are insignificant dependencies between the milk yield and climate indices. During summer, the correlation between the fat and protein concentrations with the climate indices decreased in comparison with the spring results, whereas the correlation for the milk yield increased. This methodology is suggested for studies investigating the impacts of climate variability/change on food and agriculture using short term data considering uncertainty.

The Effect of Climate Variability on Main Components of Cow Milk in Iran

The main purpose of this study is to assess the relationship between six bioclimatic indices for cattle (temperature humidity (THI), environmental stress (ESI), equivalent temperature (ESI), heat load (HLI), modified heat load (HLInew) and respiratory rate predictor(RRP)) and fundamental milk components (fat, protein, and milk yield) considering uncertainty. The climate parameters used to calculate the climate indices were taken from the NASA-Modern Era Retrospective-Analysis for Research and Applications (NASA-MERRA) reanalysis from 2002 to 2010. Cow milk data were considered for the same period from April to September when cows use natural pasture, with possibility for cows to choose to stay in the barn or to graze on the pasture in the pasturing system. The study is based on a linear regression analysis using correlations as a summarizing diagnostic. Bootstrapping is used to represent uncertainty estimation through resampling in the confidence intervals. To find the relationships between climate indices (THI, ETI, HLI, HLInew, ESI and RRP) and main components of cow milk (fat, protein and yield), multiple liner regression is applied. The least absolute shrinkage selection operator (LASSO) and the Akaike information criterion (AIC) techniques are applied to select the best model for milk predictands with the smallest number of climate predictors. Cross validation is used to avoid over-fitting. Based on results of investigation the effect of heat stress indices on milk compounds separately, we suggest the use of ESI and RRP in the summer and ESI in the spring. THI and HLInew are suggested for fat content and HLInew also is suggested for protein content in the spring season. The best linear models are found in spring between milk yield as predictands and THI, ESI,HLI, ETI and RRP as predictors with p-value < 0.001 and R2 0.50, 0.49. In summer, milk yield with independent variables of THI, ETI and ESI show the highest relation (p-value < 0.001) with R2 (0.69). For fat and protein the results are only marginal. It is strongly suggested that new and significant indices are needed to control critical heat stress conditions that consider more predictors of the effect of climate variability on animal products, such as sunshine duration, quality of pasture, the number of days of stress (NDS), the color of skin with attention to large black spots, and categorical predictors such as breed, welfare facility, and management system. This methodology is suggested for studies investigating the impacts of climate variability/change on food quality/security, animal science and agriculture using short term data considering uncertainty or data collection is expensive, difficult, or data with gaps.

Determination of subclinical metabolic disorders in transition dairy cows

Das Gesundheitsmanagement von Milchkühen hat in den vergangenen Jahren auf den landwirtschaftlichen Betrieben an Bedeutung gewonnen. Neben Präventionsmaßnahmen zur Gesunderhaltung der Tiere ist die frühzeitige und systematische Erkennung von Erkrankungen hierbei der Hauptbestandteil. Es zeigt sich vermehrt, dass vor allem Transitkühe verstärkt an Stoffwechselerkrankungen in sowohl klinischer als auch subklinischer Form erkranken. Letztere stellen ein hohes Risiko dar, zum einen weil subklinische Erkrankungen oftmals nur schwer oder gar nicht erkannt werden und zum anderen, weil sie in vielen Fällen die Grundlage für meist schwerwiegendere Folgeerkrankungen sind. In der vorliegenden Studie wird das Thema der Früherkennung von subklinischen Ketosen und der subakuten Pansenazidose behandelt. Verschiedene Methoden wurden unter praktischen Versuchsbedingungen auf ihre Tauglichkeit zur Krankheitserkennung hin geprüft. In einer ersten Studie wurde auf einem konventionellen Milchviehbetrieb ein Ketose-Monitoring bei frischlaktierenden Kühen ab Tag 3 postpartum durchgeführt. Insgesamt 15 Tiere waren an einer subklinischen Ketose erkrankt, was eine Aufkommensrate von 26% in den untersuchten Tieren bedeutete. Die Blutproben von insgesamt 24 Tieren wurden auf ihren IL-6-Gehalt untersucht. Von den untersuchten Tieren waren 14 Tiere erkrankt, 10 Tiere bildeten die gesunde Kontrollgruppe. Interleukin-6 wurde bestimmt, da dem Zytokin IL-6 in anderen Studien in Bezug auf Ketosen eine Rolle zugesprochen wurde. Die erwartete Erhöhung von IL-6 bei erkrankten Tieren konnte nicht festgestellt werden; die erkrankten Kühe zeigten vielmehr die niedrigsten IL-6 Werte der Studiengruppe. Insgesamt waren die IL-6 Konzentrationen auf einem niedrigen Niveau mit 27.2 pg/m l± 10.2. Es zeigte sich, dass die IL-6 Bestimmung im Blut hinsichtlich der Erkennung von subklinischen Ketosen nur eingeschränkt nutzbar ist. Es konnte ausschließlich eine schwache negative Korrelation zwischen Beta- Hydroxybutyrat (BHBA, Goldstandard für den Nachweis einer Ketose) und IL-6 detektiert werden. Zusätzlich zu den Blutanalysen wurde ebenfalls die tägliche Wiederkauaktivität mit dem „DairyCheck“ System bestimmt, welches kontinuierlich die charakteristischen Kaumuskelkontraktionen aufzeichnet und somit die Dauer des Wiederkäuens bestimmt werden kann. Es wurde geprüft, ob sich ketotische Tiere von nicht ketotischen Tieren hinsichtlich der täglichen Wiederkäuzeit unterscheiden. Milchkühe mit einer Ketose kauten im Schnitt 475 min/d ± 56 wieder, nach Genesung 497 min/d ± 48. Sie befanden sich somit im Durchschnitt immer unterhalb der gesunden Kontrollgruppe, welche 521 min/d ± 76 wiederkaute. Eine Korrelation zwischen der Wiederkauzeit und dem BHBA- Gehalt im Blut war nur sehr schwach ausgeprägt, nicht zuletzt da die Tiere generell eine hohe Variabilität in der Wiederkauaktivität zeigten. Bei einer weiteren Studie, ebenfalls auf einem Praxisbetrieb durchgeführt, wurde auf die Erkennung der subakuten Pansensazidose (SARA) fokussiert. Hierbei kam ein drahtloses, kommerziell verfügbares Bolussystem zum Einsatz, welches den pH Wert kontinuierlich im Retikulorumen misst. Es macht die Erkennung einer SARA auch unter Praxisbedingungen ohne invasive Methoden wie der Punktion möglich. Das Bolussystem wurde 24 Milchkühen kurz vor der Abkalbung oral eingegeben, um den pH-Wert während der gesamten Transitphase messen und überwachen zu können. Während in der Trockenstehphase nur vereinzelte SARA Fälle auftraten, erlitt ein Großteil der untersuchten Tiere in der Frühlaktation eine SARA. Auf Grundlage von pH-Werten von laktierenden Milchkühen, wurde zusätzlich eine Sensitivitätsanalyse von verschieden, bereits eingesetzten Nachweismethoden durchgeführt, um die Tauglichkeit für die SARA-Diagnostik zu untersuchen. Es handelte sich hierbei zum einen um einen SARA-Nachweis unter Heranziehung von Einzelwerten, Fress- und Wiederkäuzeiten, sowie ausgewählten Milchinhaltsstoffen und der Milchmenge. Die Analyse ergab, dass nahezu alle Nachweismethoden im Vergleich zur Langzeitmessung nur eingeschränkt zur SARA-Diagnostik nutzbar sind. In einem weiteren Teil der Studie wurde eine Kotfraktionierung bei den gleichen Tieren durchgeführt, um damit SARA-Tiere auch mittels der Kotanalyse erkennen kann. Es konnte gezeigt werden, dass zum einen die Ration einen Einfluss auf die Kotzusammensetzung hat (Trockensteherration versus Ration für Laktierende) zum anderen aber auch, dass eine SARA die Zusammensetzung des Kotes verändert. Hierfür wurden Kotproben ausschließlich von laktierenden Kühen untersucht, sodass der Einfluss der Ration ausgeschlossen werden konnte. Erhöhte Faseranteile im Kot von SARA - Kühen gaben Hinweis auf eine verminderte Verdaulichkeit. Dabei erwies sich vor allem die Hemizellulose als guter Parameter, um auf eine SARA schließen zu können. Die Versuchsbedingungen ließen es ebenfalls zu, die pH-Verläufe der Tiere in der Frühlaktation zu untersuchen. Eine Clusteranalyse von pH-Werten der ersten 12 Tage postpartum zeigte, dass die untersuchten Tiere trotz gleicher Haltungs- und Fütterungsbedingungen unterschiedliche pH-Wert Verläufe entwickelten. So gab es eine Gruppe von Milchkühen, die den pH-Wert stabil halten konnte, während die restlichen pH-Abfälle in verschiedenen Verläufen und Intensitäten aufzeigten. Es konnte ebenfalls aufgezeigt werden, dass Tiere innerhalb der Testherde unterschiedliche Schweregrade der SARA entwickelten. Auch in dieser Studie wurde deutlich, dass Tiere scheinbar unterschiedliche Möglichkeiten haben, auf ihre Umwelt zu reagieren, bzw. suboptimalen Bedingungen entgegenwirken zu können. Zusammengefasst wurden verschiedene Methoden zur Ketose- und SARA- Erkennung geprüft, von denen nur einzelne für die Praxis zu empfehlen sind. Die Variabilität der Tiere, sowohl bei der Ausprägung der Erkrankungen als auch bei den gemessenen Parametern verdeutlicht die Notwendigkeit, diese im Herden- und Gesundheitsmanagement in Zukunft stärker zu berücksichtigen.

Embryo transfer as a tool to disseminate k-casein BB genotype and improve milk clotting properties

Dairy industries are asked to be increasingly competitive and efficient. Despite the increasing trend in milk yield and protein content during the last decade genetic selection, milk coagulation ability has diminished and even if the absolute amount of cheese produced has increased, the relative cheese yield from a set amount of milk, has decreased. As casein content and variants, along with milk clotting properties (MCP) are determined to a large extent at DNA level, genetic selection and embryo transfer can provide efficacious tools to reverse this trend and achieve improvements. The aim of the proposed research was to determine how rapidly and to what extent milk coagulation properties could be improved by using embryo transfer (ET) as a tool to increase the frequency of k-casein BB genotype cattle and reducing A and E variants in an Italian Holstein herd with a low prevalence of the favourable genotype. In the effort to optimize superovulation protocols and results, synchronization of wave emergence was performed through manual transrectal ablation of the largest (dominant) ovarian follicle on days 7 or 8 of the cycle (estrus = day 0); different drugs and dosage for the superstimulation protocol were experimented trying to overcome the negative effects of stress and the perturbance of LH secretion in superovulated highly producing lactating cows and the use of SexedULTRA™ sex-sorted semen, for artificial insemination of superovulated cows was reported for the first time. The selection program carried out in this research, gave evidence and gathered empirical data of feasible genetic improvements in cheesemaking ability of milk by means of k-casein BB selection. In conclusion, in this project, selection of k-casein BB genotype markedly enhanced cheese-making properties of milk, providing an impetus to include milk coagulation traits in genetic selection and breeding programs for dairy cattle.

Development And Shelf-life Determination Of Pasteurized, Microfiltered, Lactose Hydrolyzed Skim Milk.

The segment of the world population showing permanent or temporary lactose intolerance is quite significant. Because milk is a widely consumed food with an high nutritional value, technological alternatives have been sought to overcome this dilemma. Microfiltration combined with pasteurization can not only extend the shelf life of milk but can also maintain the sensory, functional, and nutritional properties of the product. This studied developed a pasteurized, microfiltered, lactose hydrolyzed (delactosed) skim milk (PMLHSM). Hydrolysis was performed using β-galactosidase at a concentration of 0.4mL/L and incubation for approximately 21h at 10±1°C. During these procedures, the degree of hydrolysis obtained (>90%) was accompanied by evaluation of freezing point depression, and the remaining quantity of lactose was confirmed by HPLC. Milk was processed using a microfiltration pilot unit equipped with uniform transmembrane pressure (UTP) ceramic membranes with a mean pore size of 1.4 μm and UTP of 60 kPa. The product was submitted to physicochemical, microbiological, and sensory evaluations, and its shelf life was estimated. Microfiltration reduced the aerobic mesophilic count by more than 4 log cycles. We were able to produce high-quality PMLHSM with a shelf life of 21 to 27d when stored at 5±1°C in terms of sensory analysis and proteolysis index and a shelf life of 50d in regard to total aerobic mesophile count and titratable acidity.

Identification Of Corynebacterium Spp. Isolated From Bovine Intramammary Infections By Matrix-assisted Laser Desorption Ionization-time Of Flight Mass Spectrometry.

Corynebacterium species (spp.) are among the most frequently isolated pathogens associated with subclinical mastitis in dairy cows. However, simple, fast, and reliable methods for the identification of species of the genus Corynebacterium are not currently available. This study aimed to evaluate the usefulness of matrix-assisted laser desorption ionization/mass spectrometry (MALDI-TOF MS) for identifying Corynebacterium spp. isolated from the mammary glands of dairy cows. Corynebacterium spp. were isolated from milk samples via microbiological culture (n=180) and were analyzed by MALDI-TOF MS and 16S rRNA gene sequencing. Using MALDI-TOF MS methodology, 161 Corynebacterium spp. isolates (89.4%) were correctly identified at the species level, whereas 12 isolates (6.7%) were identified at the genus level. Most isolates that were identified at the species level with 16 S rRNA gene sequencing were identified as Corynebacterium bovis (n=156; 86.7%) were also identified as C. bovis with MALDI-TOF MS. Five Corynebacterium spp. isolates (2.8%) were not correctly identified at the species level with MALDI-TOF MS and 2 isolates (1.1%) were considered unidentified because despite having MALDI-TOF MS scores >2, only the genus level was correctly identified. Therefore, MALDI-TOF MS could serve as an alternative method for species-level diagnoses of bovine intramammary infections caused by Corynebacterium spp.

Metal Ions Bound To The Human Milk Immunoglobulin A: Metalloproteomic Approach.

The presence of calcium, iron, and zinc bound to human milk secretory IgA (sIgA) was investigated. The sIgA components were first separated by two-dimensional polyacrylamide gel electrophoresis and then identified by electrospray ionization-tandem mass spectrometry (ESI MS MS). The metal ions were detected by flame atomic absorption spectrometry after acid mineralization of the spots. The results showed eight protein spots corresponding to the IgA heavy chain constant region. Another spot was identified as the transmembrane secretory component. Calcium was bound to both the transmembrane component and the heavy chain constant region, while zinc was bound to the heavy chain constant region and iron was not bound with the identified proteins. The association of a metal ion with a protein is important for a number of reasons, and therefore, the findings of the present study may lead to a better understanding of the mechanisms of action and of additional roles that sIgA and its components play in human milk.

Índice de previsão de produção de leite para vacas Jersey

In Brazil the adoption of several models of cattle confinement leads to special conditions for management methods in dairy production, which can be improved by the use of technology that assures better herd management. Indexes relating environmental variables to production are applied for the prediction of milk production. The values of temperature and relative humidity, rain index, solar radiation and pasture soil temperature are generally considered potential stress agents for cows. The objective of this research was to develop an index for predicting milk production for high productivity Jersey milking cows lodged in semi confinement in tropical conditions. The experiment considered two treatments: A - the cows waited for 30 minutes prior to milking in a room with a shower associated to a fan; B - the cows did not have access to this room (control). Other than the waiting period, the cows had access to pasture. Differences in the effect of average production were not statistically significant. The analysis for studying the effect of the variables and designing the model led to a statistical model relating the variables milk production and rain index, as well as the maximum soil temperature of pasture, and milk production.

Análise do investimento em climatização para bovinos de leite em sistema de alojamento free-stall

The difficulty in adapting European dairy cows breeds in Brazil affect considerably the milk production sector. Brazilian climatic conditions are not totally favorable and the development of new tecnologies is needed for the animals express their genetic potential, as well as their best feed conversion. An economical analysis of the applied investment in the free-stall climatization equipment in dairy housing, for estimating studies related to profit, possibility of return investment as well as time for this return is necessary. The objective of this research was to evaluate the influence of climatization investment in the milk production process and analyze the economical aspect of this investment. There were used 470 high productive dairy cows with genetic and morphologic homogeneous characteristics, and analyzed in two similar periods. Investment calculations were done using Excell®. The results were satisfactory and the invested capital was proved to return to the producer in a short term, 57 days.

Composition, functional properties and sensory characteristics of Mozzarella cheese manufactured from different somatic cell counts in milk

In the present study, composition, functional properties and sensory characteristics of Mozzarella cheese produced from milk with somatic cell counts (SCC) at low (<200,000 cells/mL), intermediate (≈400,000 cells/mL) and high (>800,000 cells/mL) levels were investigated. Three batches of cheese were produced for each SCC category. The cheeses were vacuum packed in plastic bags and analysed after 2, 9, 16, 23 and 30 days of storage at 4ºC. SCC level did not affect the moisture, fat, total protein and ash content, mesophilic and psychrotrophic bacteria, and sensory parameters of Mozzarella cheese. However, meltability increased in cheese manufactured from high SCC milk. Results indicated that raw milk used to produce Mozzarella cheese should not contain high SCC (>800,000 cells/mL) in order to avoid changes in the functional properties of the Mozzarella cheese.

Morfometria e distribuição de leite alveolar e cisternal na glândula mamária de vacas Holandesa e Girolanda

Estudaram-se a distribuição do leite nas porções alveolar e cisternal e a morfometria da glândula mamária de 14 vacas, sete da raça Holandesa, grupo H, e sete da Girolanda, grupo G, com ordem e dias de lactação semelhantes, ordenhadas duas vezes ao dia. A produção de leite foi mensurada diariamente, enquanto as frações de leite cisternal e alveolar foram medidas uma vez por semana. Durante o experimento, também foram realizadas medidas de ultrassonografia e morfometria da glândula mamária. As vacas Girolandas apresentaram úberes e tetos maiores que as vacas Holandesas. Diâmetro, espessura da parede e comprimento do canal do teto, medidos por imagens de ultrassom, também foram maiores nas Girolandas, mas a área de cisterna da glândula mamária foi maior nas Holandesas. Não houve diferença na produção de leite e nas frações de leite cisternal e alveolar entre as raças.