A review of vitamin D and its importance to the health of dairy cattle.

This Research Reflection short review will discuss vitamin D metabolism, its role in nutrition, disease prevention, and welfare of dairy cattle, as well as its toxicity. Vitamin D is an important fat-soluble vitamin. However, some researchers regard it as a hormone due to its function in the organism. Its role is not limited just to Ca homoeostasis and bone metabolism but is also associated with immunity. In dairy cattle it is known for preventing milk fever. Cows can acquire vitamin D in many ways for example through feed, parenteral injections or through UVB irradiation from the sun or artificial lighting. The vitamin D in feed can either be plant-/ fungi- based ergocalciferol or animal-based cholecalciferol. There is currently only one registered feed vitamin D supplement for cattle in the European Union and it is cholecalciferol. Animals can also synthesize their own vitamin D when 7-dihydrocholesterol in the skin is irradiated with UVB light resulting in cholecalciferol production. Despite its importance, many cattle are deficient in vitamin D due to inadequate supplementation or insufficient sun exposure. In a study performed at the Veterinary Faculty in Slovenia 12 high producing Holstein Friesian cows at a commercial dairy farm were blood tested for vitamin D status for three succeeding months and all but one were vitamin D insufficient in all testings. The cows were not exposed to direct sunlight and the content of vitamin D3 in feed was <400 IU/kg dry matter, which is less than half of the NRC (2001) recommendation. Deficiency can also occur due to diseases affecting the gastrointestinal tract, such as paratuberculosis, which lower the absorptive capacity of the gut. Vitamin D can be toxic if cows are over-supplemented or consume large quantities of plants like Trisetum flavescens, which contain an active form of vitamin D-calcitriol or its glycosides, that are activated by digestion in the rumen.

Management practices that influence the welfare of calves on small family farms.

In this Research Reflection we review management practices in small family farms with less than 100 cows. Small farms represent the majority of farms in the EU and the world, and they are of great importance for the economy of a country. On cattle farms, the welfare of calves is of primary importance for the profitability of the herd, and poor management is one of the main factors influencing calf health and survival. Data on the risk factors for calf welfare issues in small-scale farms are limited. For this purpose, the literature data from six world countries were presented and compared, including Serbia and Slovenia where a survey related to the issue was carried out within the COST Action FA1308, DairyCare. Some practices within the following areas in calf management were considered: calving management, care for new-born calves, use of painful procedures, colostrum management, cow-calf separation, calf feeding, weaning, calf housing, and general monitoring. In each of the countries, the health and welfare of calves are threatened by some omissions in rearing practices and the major are related to the new-born calf management, the feeding and watering management, and the application of hygienic measures. Many farmers are well aware of the importance of proper calf rearing; others would need more incentive to improve calf management. Each country should pay attention to the education of farmers about the most common deficiencies in calf management.

Welfare assessment in dairy cows using hair cortisol as a part of monitoring protocols.

Welfare of dairy cows can be assessed using welfare assessment protocols consisting of resource, management and animal-based measures. Welfare Quality® Assessment Protocol is one of the best-known protocols, which depends almost entirely on animal-based measures. To gain more objective and rapid welfare assessment, new techniques have been developed to measure welfare of animals, such as hair cortisol concentration. As cortisol is released in response to stress, it has long been used as a biomarker of stress in animals. While the precise mechanism of cortisol incorporation into hair is unknown, hair cortisol concentration seems to be a marker of long-term systemic cortisol concentration. Hair cortisol is, therefore, a potential marker of chronic stress and is not likely to be affected by acute stress. Studies on cattle show connections between hair cortisol concentration and factors such as pregnancy, parity, diseases, ectoparasites, body condition score, environmental changes, stocking density and milk yield. Hair cortisol concentration appears to be affected by time of sampling, cow age and breed, UV radiation, season, body region of sampled hair and hair colour. Its concentration also depends on sampling and analytical methods. Hair cortisol is a promising non-invasive tool to evaluate welfare of dairy cows, however, more research is needed to determine the extent of effects on its concentration and the appropriate method of sampling and analysis. Correlations between Welfare Quality® Assessment Protocol scores and pooled hair cortisol concentrations have not yet been found, and more research is needed with larger sample size, a standardized protocol of hair sampling, processing and analysis. With proper attention to detail, hair cortisol levels in pooled hair samples might come to be used as a reliable indicator of dairy animal welfare.

Coronaviruses in cattle.

Bovine coronaviruses are spread all over the world. They cause two types of clinical manifestations in cattle either an enteric, calf diarrhoea and winter dysentery in adult cattle, or respiratory in all age groups of cattle. The role of coronaviruses in respiratory infections is still a hot topic of discussion since they have been isolated from sick as well as healthy animals and replication of disease is rarely successful. Bovine coronavirus infection is characterised by high morbidity but low mortality. The laboratory diagnosis is typically based on serological or molecular methods. There is no registered drug for the treatment of virus infections in cattle and we are limited to supportive therapy and preventative measures. The prevention of infection is based on vaccination, biosecurity, management and hygiene. This paper will cover epidemiology, taxonomy, pathogenesis, clinical signs, diagnosis, therapy, economic impact and prevention of coronavirus infections in cattle.

Congenital hypotrichosis in a Simmental cross heifer.

BACKGROUND: Congenital hypotrichosis is defined as a less than normal amount of hair, obvious at birth or in the first weeks of life. Causes are nongenetic or genetic. OBJECTIVES: To describe the clinical presentation and histological features of a novel form of hypotrichosis in a heifer. ANIMAL: A 15-month-old Simmental cross-bred heifer was presented with a history of progressive hair loss, which started at four months of age and progressed to severe generalized hypotrichosis. METHODS AND MATERIALS: Anamnestic data, detailed clinical examination, haematological investigation, selected biochemistry profile, RT-PCR and ELISA for bovine viral diarrhoea virus did not suggest a cause for the hypotrichosis; skin samples were examined histologically. RESULTS: Histopathology of the skin showed a 50% reduction in the number of hair follicles and dysplastic hair follicles; these findings led to a diagnosis of congenital hypotrichosis. The distribution of the hair and some of the histopathological characteristics were potentially consistent with viable hypotrichosis. A marked reduction in follicle density, hypertrophy of sebaceous glands, hypoplasia of arrector pili muscles and moderate acanthosis with hyperkeratosis, which have not been described previously in viable hypotrichosis or other forms of congenital hypotrichosis, were also observed. CONCLUSION AND CLINICAL SIGNIFICANCE: This case may represent a novel form of congenital hypotrichosis.

Detection of tetracycline and other antimicrobial residues in milk from cows with clinical mastitis treated by combination therapy.

The purpose of this study was to determine tetracycline and other antimicrobial residues in milk from dairy cows treated for clinical mastitis. Data on cow health and drugs used were obtained from the farm veterinarians. A milk sample from each affected udder quarter of each cow was taken for bacterial identification before treatment. All 35 cows in the study were treated with an intramammary product containing tetracycline, neomycin, bacitracin, and prednisolone (tetracycline injector) and also simultaneously with other drugs via the intramammary or parenteral route. The withhold period of the 'tetracycline product' was eight milkings but, in all cases, longer withhold periods were prescribed due to off-label application of additional combinations of drugs. Milk samples from treated udder quarters, taken two milkings before and two after the prescribed withhold period, were analysed for antimicrobial residues. Additionally, milk samples were taken from untreated healthy udder quarters to check if any crossover of drugs had occurred. Three screening tests were used for antimicrobial detection. In 15 (42·9%) cows the milk samples from the treated infected quarters contained tetracycline residues above the maximum residue limit (MRL) after the prescribed withhold period and, in two cases (5·7%) the same was true for neomycin and cefquinome residues. Cephalexin above the MRL was detected in only one case (2·9%). Beta-lactams did not exceed the MRL after the prescribed withhold period. Antimicrobial residues were not detected in milk samples from untreated quarters. Cow condition including parity, decreased milk production and severity of mastitis significantly influenced the excretion of antimicrobials in milk (P ≤ 0·05). No significant differences were found between the tetracycline positive and tetracycline negative cows regarding the bacteria involved, combination antimicrobial therapy, or treatment interval and duration.

Analytical and Clinical Interference of Sample Hemolysis in Evaluating Blood Biochemical and Endocrine Parameters in Cows.

Hemolysis is a common cause of errors in laboratory tests as it affects blood parameters and leads to a positive or negative bias. This study aims to examine the relationship between the level of hemolysis (expressed as cell-free hemoglobin concentration, g/L) and the variability of metabolic and endocrine parameters and to determine the threshold level of hemolysis that causes an analytically and clinically significant bias for the twenty most frequently examined blood parameters in cows. Paired blood samples of 10 mL each were obtained from 30 cows. One was subjected to mechanical trauma and plasma was extracted directly from the other. Hemolyzed and non-hemolyzed samples from the same animal were mixed to obtain final samples with cell-free hemoglobin concentrations of 0, 1, 2, 4, 6, 8, and 10 g/L. Metabolic and endocrine parameters were measured in the samples and their deviation and the linear equation between the level of hemolysis and the deviation were determined. The following threshold values of hemolysis were determined, which correspond to the acceptable analytical (lower value) and clinical (upper value) levels of parameter variability: BHB 0.96 and 4.81; NEFA 0.39 and 3.31; GLU 0.38 and 3.90; ALB 1.12 and 6.11; TPROT 1.40 and 6.80; UREA 6.62 and 20.1; TBIL 0.75 and 5.65; AST 0.11 and 2.18; GGT 1.71 and 8.90, LDH 0.01 and 0.11, ALP 0.97 and 2.95; TGC 1.56 and 15.5; CHOL 1.29 and 8.56; Ca 5.68 and 25.7; P 0.57 and 8.43; Mg 1.10 and 8.47; INS 1.15 and 3.89; T3 8.19 and 15.6; T4 8.97 and 18.5; and CORT 2.78 and 11.22 g/L cell-free hemoglobin. Three decision levels are available for each metabolic and endocrine parameter: if hemolysis is below the lower (analytical) threshold value, results can be reported without restriction; if hemolysis is between the lower and upper thresholds, the results can be issued with guidance in the form of corrective linear equations; and if hemolysis is above the upper (clinical) threshold, the results and sample must be discarded. This method contributes to an optimal approach to hemolysis interference with metabolic profile parameters in blood samples from cows.

Evaluation of combined high-efficiency DNA extraction and real-time PCR for detection of Mycobacterium avium subsp. paratuberculosis in subclinically infected dairy cattle: comparison with faecal culture, milk real-time PCR and milk ELISA.

BACKGROUND: Johne's disease is caused by Mycobacterium avium subsp. paratuberculosis (Map) and it is one of the most important diseases in cattle worldwide. Several laboratory tests for Map detection are available; however, these are limited by inadequate sensitivity and specificity when used in subclinically infected populations. To identify Map shedders in subclinically infected cattle, we used a new, high-yield method for DNA-extraction from Map in faeces combined with quantitative real-time PCR (qPCR) for amplification of the insertion sequence IS900 of Map (HYDEqPCR). Evaluation of HYDEqPCR was carried out in comparison with faecal culture, milk qPCR, and milk enzyme-linked immunosorbent assay (ELISA), on 141 faecal and 91 milk samples, from 141 subclinically infected dairy cattle. RESULTS: The qPCR proved to be highly sensitive, with a detection limit of 2 IS900 DNA copies/µl in 67 % of the reactions. It also showed 100 % specificity, as determined from 50 Map and non-Map strains, and by the sequencing of qPCR amplicons. The detection limit of HYDEqPCR was 90 Map/g Map-spiked faeces, which corresponds to 2.4 colony forming units/g Map-spiked faeces, with an estimated efficiency of 85 % (±21 %). When tested on the field samples, HYDEqPCR showed 89 % of the samples as positive for Map, whereas faecal culture, milk qPCR, and milk ELISA detected 19 %, 36 % and 1 %, respectively. Fisher's exact tests only show statistical significance (p ≤0.05) for the correlation between HYDEqPCR and faecal culture. The agreement between HYDEqPCR and milk qPCR and milk ELISA was poor, slight, and non-significant. CONCLUSIONS: This study highlights the advantages of HYDEqPCR for detection of Map in subclinically infected populations, in comparison with faecal culture, milk qPCR and milk ELISA. HYDEqPCR can detect low-level Map shedders that go undetected using these other methods, which will thus underestimate the proportions of Map-shedders in herds. Identification of these shedding animals is extremely important for prevention of the spread of Map infection in an animal population. Due to the relatively high sensitivity and specificity of HYDEqPCR, it can be applied to test for Map at the herd or individual level, regardless of animal age or production stage. HYDEqPCR will allow early detection and control of Map in any population at risk.

The Influence of Energy Balance, Lipolysis and Ketogenesis on Metabolic Adaptation in Cows Milked Twice and Three Times Daily.

Increasing milk production requires increasing milking frequency (MF) from two times daily (2X) to three (3X) or more. High milk production leads to negative energy balance (NEB) and homeorhesis, characterized by lipolysis, ketogenesis, and endocrinological changes. The relationship among energy balance (EB), lipolysis, and ketogenesis with endocrine and metabolic parameters in blood of cows milked 2X and 3X daily was studied. Holstein Friesian cows milked 2X (n = 45) and 3X (n = 45) were analyzed, with approximately 50% of cows in each group in positive EB (PEB) and 50% in NEB. After determining EB, blood samples were collected from all cows and blood serum was analyzed for non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), glucose (GLU), cholesterol (CHOL), triglycerides (TGC), total bilirubin (TBIL), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), calcium (Ca), inorganic phosphate (P), total protein (TPROT), albumin (ALB), urea, insulin (INS), T3, T4, and cortisol (CORT), and the RQUICKIBHB index of insulin resistance was calculated. Cows milked 3X in NEB represent a special cluster that partially overlaps with cows milked 2X in NEB and has no contact points with cows in PEB. Cows milked 3X had higher levels of NEFA, BHB, AST, GGT, TBIL, and CORT and lower levels of GLU, Ca, INS, and T4. Cows milked 3X in NEB had higher levels of NEFA, BHB, AST, GGT, TBIL, and CORT and lower levels of GLU, CHOL, TGC, TPROT, P, INS, RQUICKIBHB, and T3 compared with cows milked 2X in NEB and cows in PEB. In cows milked 3X, lipolysis and ketogenesis were much more prominent, and EB levels were lower, implying a pronounced shift in homeorhesis. Metabolic and endocrinology parameters were determined mainly by the values of EB, NEFA, and BHB in cows milked 3X in NEB compared with other categories of cows. The results confirm the peculiarity of metabolic adaptation in cows with increased MF, characterized not only by differences in the concentration of metabolites but also in their interactions.

Identifying "Vulnerable Agricultural Populations" at Risk for Occupational Injuries and Illnesses: A European Perspective.

Objectives: A common understanding of the term "vulnerable populations" in the European agricultural context is needed. Therefore, the purpose of this paper was to identify vulnerable populations working in European agriculture.Methods: Participatory dialogue with members of the Safety Culture and Risk Management in Agriculture (SACURIMA) network was used to identify and build consensus on the major vulnerable groups of people who work in European agriculture.Results: Five groups of vulnerable people were identified, which included: (1) foreign-born farmworkers (including both immigrants and refugees); (2) migrant and seasonal farmworkers; (3) beginning farmers (those with <5 years of experience); (4) farm families (including women, children, and older adults), and (5) farmers and farmworkers who have physical, mental health, or intellectual disabilities.Conclusion: By developing a consistent understanding of vulnerable populations working in European agriculture, we can promote consistency in health and safety messaging, measurement of health and safety constructs, and implementation and dissemination of health and safety programs and information.

Review of Infections With Bovine Herpesvirus 1 in Slovenia.

In the 1950s, infectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV) disease was clinically detected and documented in cattle for the first time in Slovenia. The bovine herpes virus 1 (BoHV-1) was confirmed several times from infected herds by virus isolation on cell cultures. To keep the IC virus-free, high biosecurity measures were introduced. Before entering the IC, all calves are serologically tested and quarantined. Bulls in Slovenian insemination centres (IC) have been negative for IBR /IPV infection since 1979. From 1985 to 1991, few large-scale studies of the prevalence of IBR/IPV were carried out. In 1985, a high percentage (56.9%) of serologically positive animals were found in large state farms with Holstein Friesian cattle. Epidemiological studies in farm with bulls' mother herds were also carried out in the farms with Simmental and Brown cows. Antibodies against BoHV-1 were detected in the serum of 2.3% of Brown cattle and 3.5% of Simmental cattle. In the year 2000, 3.4% of bulk tank milk samples from 13,349 dairy farms were detected BoHV-1 antibodies positive. The highest percentage of positive animals was found in regions with an intensive grazing system (6.2% positive) and the lowest percentage in the east part of Slovenia (0.9% positive) on farms with mostly Simmental cattle. In 2006, a total 204,662 sera of cattle older than 24 months were tested for the presence of BoHV-1 antibodies and positive cattle were detected in 3.6% of tested farms. These farms kept 34,537 animals that were potential carriers of the BoHV-1. Most of the positive farms kept Holstein Friesian cattle, descendants from the state-owned farms, which were privatised or closed after 1990. In 2015, the Administration of the Republic of Slovenia for Food Safety, Veterinary and Plant Protection issued a rule that describes the conditions for granting and maintaining the status of BoHV-1 free holdings. The rule provides a voluntary control programme for breeders who want to obtain BoHV-1 free status and are willing to cover all the cost of acquiring and maintaining that status. There has been very little response from breeders.

The Principles of the Voluntary Programme for the Control and Elimination of Bovine Viral Diarrhoea Virus (BVDV) From Infected Herds in Slovenia.

In Slovenia, the control of bovine viral diarrhoea virus (BVDV) infections started in 1994. Since 2014, a voluntary programme has been running according to the national rules that prescribe the conditions for recognising, acquiring, and maintaining a BVDV-free status for an individual herd. The principle is based on periodical laboratory testing and preventive measures that need to be strictly implemented in a herd. Between 2014 and 2020, a total of 348 herds were included in BVDV antibody testing, and 25.0% of tested herds were detected to be BVDV antibody positive. To recognise the BVDV-free status of the herd, the breeder should provide two consecutive tests with intervals of at least 6 months in all animals in the age from 7 to 13 months, with negative results for BVDV antibodies in ELISA. The BVDV-free status of the herd can be maintained by implementing preventive measures and can be renewed each year with one laboratory test in the age group of animals from 7 to 13 months for antibodies in ELISA. During the 7 years of the voluntary programme, 236 herds were included in the detection of BVDV in individual herds by real-time RT-PCR method and the elimination of positive animals from herds. In 71 (31.3%) herds, at least one BVDV-positive animal was detected, with the identification of a total of 267 persistently infected (PI) animals, representing an average of 2.9% of tested animals. The cost of testing for an average herd, recognised as BVDV-negative, and maintaining its BVDV-free status within the implemented voluntary programme, was €97.64/year, while for the average positive herd, the laboratory costs for elimination of BVDV were €189.59/year. Only limited progress towards eradication at the national level has been achieved in Slovenia since 2014.

Minimal Erythema Dose Determination in Holstein Friesian Cattle.

Cattle on pasture are continuously exposed to solar UV radiation, which has been associated with biological effects such as sunburn, photosensitization, squamous cell carcinoma, and cutaneous vitamin D3 production. The minimal erythema dose (MED) required to produce first-degree sunburn (erythema) is poorly researched in cattle. Since cattle are naturally covered with dense hair coats, the MED is influenced by the UV protection offered by the hair. The objective of this study was to determine the MED on intact-hair-covered (MED-H) and shaved white skin (MED-S) of Holstein Friesian cattle. Twenty-one Holstein Friesian cows and heifers were MED tested using a narrowband UV-B LED light (peak irradiance at 292 nm) on eight hair-covered and eight shaved areas over white skin previously unexposed to direct sunlight. Erythema was visually assessed after 24 h. The mean MED-H and MED-S were 5,595 and 329 J/m2, respectively. Heifers had a higher MED-H compared to cows, 7,600 and 4,969 J/m2, respectively. The mean UV transmittance of white cattle hair was 6.7%. MED-H was correlated with hair length (Spearman's rho = 0.76). A linear regression model showed that each millimeter of hair coat length increased the MED-H by 316 J/m2. In conclusion, this study provides a MED testing protocol for cattle and reports standardized values of MED for cattle on intact-hair-covered and shaved areas.

Corrigendum: Overview of Slovenian Control Programmes for Selected Cattle Diseases, Listed Under Category C, D or E of the European Animal Health Law.

[This corrects the article DOI: 10.3389/fvets.2021.674515.].

Overview of Slovenian Control Programmes for Cattle Diseases Not Regulated by the European Union.

The European Union (EU) regulates the control of cattle diseases listed in categories A and B of the Animal Health Law (AHL). However, the control of other cattle diseases that have no, or limited EU regulation, is left to each member state. Slovenia has five control programmes (CPs) for non-EU regulated cattle diseases: bovine viral diarrhoea (BVD), infectious bovine rhinotracheitis (IBR), enzootic bovine leukosis (EBL), bluetongue and anthrax. Two (IBR and BVD) are voluntary and the others (EBL, anthrax and bluetongue) are compulsory. The three compulsory CPs are funded by the government. All the CPs are run by the government and laboratory tests are performed by the National Veterinary Institute. The rules for the CPs are laid down in Slovenian legislation. In addition, there is a national directive for the control of salmonellosis. Both BVD and IBR are endemic and have CPs based on increased biosecurity, testing and culling or vaccination, financed by the animal owners. Slovenia has been officially free of EBL since 2005 and carries out surveillance based on serological testing of a representative number of herds and inspection of carcasses at slaughter or necropsy. Vaccination is the main disease control measure for anthrax (sporadic) and bluetongue (currently perceived free-vaccination since 2017). Lack of motivation of farmers to participate in voluntary disease CPs and to implement and follow strict biosecurity measures are the most pressing issues in improving the health status of Slovenian cattle. An overview of the existing CPs and the circumstances leading to their implementation are presented.

The Correlation between Extracellular Heat Shock Protein 70 and Lipid Metabolism in a Ruminant Model.

Metabolic stress in early lactation cows is characterized by lipolysis, ketogenesis, insulin resistance and inflammation because of negative energy balance and increased use of lipids for energy needs. In this study the relationship between lipid metabolite, lipid-based insulin resistance, and hepatocyte functionality indexes and tumor necrosis factor alpha (TNF-α) with extracellular heat shock protein 70 (eHsp70) was investigated. The experiment included 50 cows and all parameters were measured in blood serum. In cows with a more pronounced negative energy balance, the following was determined: a higher concentration of eHsp70, TNF-α, non-esterified fatty acid (NEFA), beta-hydroxybutyrate (BHB), NEFA to insulin and NEFA to cholesterol ratio and lower concentration of cholesterol, very low-density lipoproteins (VLDL), low density lipoproteins (LDL) and liver functionality index (LFI). The eHsp70 correlated negatively with the values of cholesterol, VLDL, LDL, and triglycerides, while correlated positively with the level of NEFA and BHB. A higher concentration of eHsp70 suggests the development of fatty liver (due to a higher NEFA to cholesterol ratio and lower LFI) and insulin resistance (due to a lower revised quantitative insulin sensitivity check index RQUICKI-BHB and higher NEFA to insulin ratio). The eHsp70 correlated positively with TNF-α. Both TNF-α and eHsp70 correlated similarly to lipid metabolites. In cows with high eHsp70 and TNF-α values we found higher concentrations of NEFA, BHB, NEFA to insulin and NEFA to cholesterol ratio and a lower concentration of triglycerides and VLDL cholesterol compared to cows that had only high TNF-α values. Based on the positive correlation between eHsp70 and TNF-α, their similar relations, and the additional effect of eHsp70 (high TNF-α + eHsp70 values) on lipid metabolites we conclude that eHsp70 has pro-inflammatory effects implicating lipolysis, fatty liver, and fat tissue insulin resistance.

Domestic animal proteomics in the 21st century: A global retrospective and viewpoint analysis.

Animal production and health are of significant economic importance, particularly regarding the world food supply. Animal and veterinary sciences have evolved immensely in the past six decades, particularly in genetics, nutrition, housing, management and health. To address major challenges such as those posed by climate change or metabolic disorders, it is of utmost importance to use state-of-the-art research tools. Proteomics and the other post-genomic tools (transcriptomics or metabolomics) are among them. Proteomics has experienced a considerable development over the last decades. This brought developments to different scientific fields. The use and adoption of proteomics tools in animal and veterinary sciences has some limitations (database availability or access to proteomics platforms and funding). As a result, proteomics' use by animal science researchers varies across the globe. In this viewpoint article, we focus on the developments of domestic animal proteomics over the last decade in different regions of the globe and how the researchers have coped with such challenges. In the second part of the article, we provide examples of funding, educational and laboratory establishment initiatives designed to foster the development of (animal-based) proteomics. International scientific collaboration is a definitive and key feature in the development and advancement of domestic animal proteomics. SIGNIFICANCE: Animal production and health are very important for food supply worldwide particularly as a source of proteinaceous foods. Animal and veterinary sciences have evolved immensely in the last decades. In order to address the major contemporary challenges facing animal and veterinary sciences, it is of utmost importance to use state-of-the-art research tools such as Proteomics and other Omics. Herein, we focus on the major developments in domestic animal proteomics worldwide during the last decade and how different regions of the world have used the technology in this specific research field. We address also major international efforts aiming to increase the research output in this area and highlight the importance of international cooperation to address specific problems inherent to domestic animal proteomics.

Biochemical Bone Markers During the Transition Period Are Not Influenced by Parenteral Treatment With a High Dose of Cholecalciferol but Can Predict Milk Fever in Dairy Cows.

Despite being studied extensively, there are still many knowledge gaps in milk fever prevention and it is still a prevalent disease. Various interventions have been used in its prevention; however, none has proven to be entirely effective. The study aimed to assess the effectiveness of high dose vitamin D3 parenteral (intramuscularly) administration and the mechanism of its action by studying blood minerals and biochemical bone markers. Further, we assessed the potential of biochemical bone markers, measured in the close-up dry period, as predictors of clinical milk fever after calving. The study was conducted on 56 high yielding, clinically healthy dairy cows, before their 4th or higher lactation. They were divided into three groups based on season (summer and winter) and administration (vitamin D). The winter group was considered as the control group. Cows (n = 13) were parenterally administered a single dose of 10 million IU of vitamin D3 (DUPHAFRAL® D3) ranging between 10 and 2 days before calving (median = 3 days). Each cow was blood sampled once during four sampling period ranges: ~1 month before calving, 10 to 2 days before calving, 12-48 h after calving and 10-20 days after calving. The samples were analyzed for blood minerals, bone specific alkaline phosphatase (bALP) and C-terminal telopeptide of type I collagen (CTx), alkaline phosphatase, and estradiol. Values were compared between samplings and groups. A receiver operating characteristic (ROC) analysis and logistic regression were used to assess the diagnostic accuracy of biochemical bone markers in predicting milk fever. In this study high dose vitamin D3 supplementation did not statistically reduced the incidence of milk fever (milk fever incidences were 15.4, 39.1, and 25% in the vitamin D, winter and summer groups, respectively). A significant effect of vitamin D3 administration on blood minerals or biochemical bone markers was not found at any sampling. We found that the use of biochemical bone markers in the close-up dry period to predict clinical milk fever was applicable only in the winter (housed) group. The area under the curve (AUC) for bALP was 0.804 and 0.846 for CTx using ROC analysis. The bALP curve had the best ratio at the cut-off point 13.85 U/L with 90% sensitivity and 64.3% specificity. While CTx had the ratio of 90% sensitivity and 78.6% specificity at the cut-off point 0.149 ng/mL. Close-up dry dairy cows with CTx ≥0.121 ng/mL had a 3.8 times higher chance of succumbing to milk fever. We were unable to prove that high dose vitamin D3 parenteral administration is a viable technique for milk fever prevention. Biochemical bone markers are a promising tool for predicting milk fever; however, further studies are needed to confirm their clinical use.

Melatonin treatment in winter and spring and reproductive recovery in Sarda breed sheep.

This study aimed to evaluate the effect of melatonin treatment carried out between late winter and early spring on reproductive response in Sarda breed sheep and whether the photo-refractoriness can influence this reproductive response. The study was conducted on 3200 adult ewes, aged 3-6years old, with body condition score (BCS) 2.5-4.0, from 16 commercial sheep farms in Northern Sardinia. In each farm 200 animals were enrolled and subdivided into 2 groups (n=100 each): Group M (treated with one 18mg melatonin implant), and group C (untreated). The melatonin treatments were performed on 10th February; 10th March; 10th April and on 10th May each time in 4 different randomly selected farms. Adult males, treated with 3 melatonin implants 1 week before females, were introduced in each flock 35days after ewes' treatment, and removed after 45days of cohabitation with females. Pregnancy was determined by transabdominal ultrasonography examination between 45th and 90th day after ram introduction. Data showed that melatonin treatment increased the fertility rate significantly (P<0.05), with the higher fertility rate in the ewes treated in April and May. The average time in days from male introduction to lambing was shorter in treated than in control ewes (P<0.05). Further, at the 160th and 170th day after male introduction the group M showed a higher number of lambed ewes compared to C (P<0.01). This effect was observed at 180th and 190th days after ram introduction, also, but with lower significance (P<0.05). In conclusion, melatonin treatment improved reproductive efficiency and advanced breeding season in Sarda sheep, especially when ewes were treated in spring.

Epidemiology of taeniosis/cysticercosis in Europe, a systematic review: Western Europe.

BACKGROUND: Taenia solium and Taenia saginata are zoonotic parasites of public health importance. Data on their occurrence in humans and animals in western Europe are incomplete and fragmented. In this study, we aimed to update the current knowledge on the epidemiology of these parasites in this region. METHODS: We conducted a systematic review of scientific and grey literature published from 1990 to 2015 on the epidemiology of T. saginata and T. solium in humans and animals. Additionally, data about disease occurrence were actively sought by contacting local experts in the different countries. RESULTS: Taeniosis cases were found in twelve out of eighteen countries in western Europe. No cases were identified in Iceland, Ireland, Luxembourg, Norway, Sweden and Switzerland. For Denmark, Netherlands, Portugal, Slovenia, Spain and the UK, annual taeniosis cases were reported and the number of detected cases per year ranged between 1 and 114. Detected prevalences ranged from 0.05 to 0.27%, whereas estimated prevalences ranged from 0.02 to 0.67%. Most taeniosis cases were reported as Taenia spp. or T. saginata, although T. solium was reported in Denmark, France, Italy, Spain, Slovenia, Portugal and the UK. Human cysticercosis cases were reported in all western European countries except for Iceland, with the highest number originating from Portugal and Spain. Most human cysticercosis cases were suspected to have acquired the infection outside western Europe. Cases of T. solium in pigs were found in Austria and Portugal, but only the two cases from Portugal were confirmed with molecular methods. Germany, Spain and Slovenia reported porcine cysticercosis, but made no Taenia species distinction. Bovine cysticercosis was detected in all countries except for Iceland, with a prevalence based on meat inspection of 0.0002-7.82%. CONCLUSIONS: Detection and reporting of taeniosis in western Europe should be improved. The existence of T. solium tapeworm carriers, of suspected autochthonous cases of human cysticercosis and the lack of confirmation of porcine cysticercosis cases deserve further attention. Suspected cases of T. solium in pigs should be confirmed by molecular methods. Both taeniosis and human cysticercosis should be notifiable and surveillance in animals should be improved.