Case-control study on factors associated with a decreased milk yield and a depressed health status of dairy herds in northern Germany.

BACKGROUND: In the past years, it became apparent that health status and performance differ considerably within dairy farms in Northern Germany. In order to obtain clues with respect to possible causes of these differences, a case-control study was performed. Case farms, which showed signs of health and performance problems, and control farms, which had none of these signs, were compared. Risk factors from different areas such as health management, housing, hygiene and nutrition were investigated as these are known to be highly influential. The aim of this study was to identify major factors within these areas that have the strongest association with health and performance problems of dairy herds in Northern Germany. RESULTS: In the final model, a lower energy density in the roughage fraction of the diet, more pens with dirty lying areas and a low ratio of cows per watering spaces were associated with a higher risk for herd health problems. Moreover, case farms were affected by infections with intestinal parasites, lungworms, liver flukes and Johne's Disease numerically more often than control farms. Case farms more often had pens with raised cubicles compared to the deep bedded stalls or straw yards found in control farms. In general, the hygiene of the floors and beddings was worse in case farms. Concerning nutrition, the microbiological and sensory quality of the provided silages was often insufficient, even in control farms. Less roughage was provided to early lactating cows and the feed was pushed to the feeding fence less frequently in case farms than in control farms. CONCLUSIONS: The results show that milk yield and health status were associated with various factors from different areas stressing the importance of all aspects of management for good animal health and performance. Moreover, this study confirmed well-known risk factors for health problems and performance losses. These should better be taken heed of in herd health management.

Diversity of Clostridium perfringens toxin-genotypes from dairy farms.

BACKGROUND: Clostridium (C.) perfringens is the causative agent of several diseases in animals and humans, including histotoxic and enteric infections. To gain more insight into the occurrence of its different toxin-genotypes in dairy herds, including those toxin genes previously associated with diseases in cattle or humans, 662 isolates cultivated from feces, rumen content and feed collected from 139 dairy farms were characterized by PCR (detecting cpa, cpb, iap, etx, cpe, and both allelic variants of cpb2). RESULTS: Isolates from feces were assigned to type A (cpa positive, n = 442) and D (cpa and etx positive, n = 2). Those from rumen content (n = 207) and feed (n = 13) were all assigned to type A. The consensus and atypical variants of the cpb2 gene were detected in 64 (14.5 %) and 138 (31.22 %) of all isolates from feces, and 30 (14.5 %) and 54 (26.1 %) of all isolates from rumen content, respectively. CONCLUSION: Both allelic variants of cpb2 occurred frequently in animals without signs of acute enteric disease, whereby the atypical variant dominated. Five (0.8 %) of all type A isolates were positive for the cpe gene. Therefore, the present study indicates that dairy cows are no primary source for potentially human pathogenic enterotoxin gene positive strains.

Detection of Clostridium botulinum neurotoxin genes (A-F) in dairy farms from Northern Germany using PCR: A case-control study.

Classical botulism in cattle mainly occurs after ingestion of feed contaminated with preformed toxin. In 2001 a form of botulism ("visceral botulism") was postulated to occur after ingestion of Clostridium (C.) botulinum cells or spores, followed by colonization of the intestine, and local production of botulinum neurotoxin (BoNT) causing chronic generalized disease. To verify the potential role of C. botulinum in the described syndrome, a case-control study was conducted, including 139 farms. Fecal samples, rumen content, water and silage samples were collected on each farm. Real time BoNT gene PCR assays were conducted after enrichment in RCM (Reinforced Clostridial Medium) at 37 °C and conventional PCRs after enrichment in MCM (Modified Cooked Meat Medium) at 30 °C. Furthermore, a direct detection of BoNT genes without prior enrichment was attempted. BoNT A, B, C, D, E and F genes were detected in animal samples from 25 (17.99%), 3 (2.16%), 0 (0.0%), 2 (1.44%), 1 (0.72%), and 3 (2.16%) farms, respectively. Eleven feed samples were positive for BoNT A gene. By enrichment a significant increase in sensitivity was achieved. Therefore, this should be an essential part of any protocol. No significant differences regarding BoNT gene occurrence could be observed between Case and Control farms or chronically diseased and clinically healthy animals within the particular category. Thus, the postulated form of chronic botulism in cows could not be confirmed. This study supports the general opinion that C. botulinum can occasionally be found in the rumen and intestine of cows without causing disease.

[Selective dry cow therapy on dairy farms in Rhineland-Palatinate, Saarland and Hesse, Germany - a survey among farmers]. / Selektives antibiotisches Trockenstellen bei Milchkühen in Rheinland-Pfalz, dem Saarland und Hessen ­ eine Umfrage unter Landwirten.

OBJECTIVE: With the Regulation (EC) 6/2019, antibiotic drying off of the entire dairy herd is no longer permissible. Hence, it is necessary to establish selective antibiotic drying off (SDCT: Selective Dry Cow Therapy) in dairy herds. With the publication of the PraeRi study in 2020, systematic data for the implementation of SDCT on farms became available for several German states. For Rhineland-Palatinate, Saarland and Hesse this type of information is only available from individual projects. Therefore, the aim of this survey was to increase the knowledge concerning the implementation of SDCT in dairy farms located in these states. MATERIAL AND METHODS: An online questionnaire was sent via newsletters to farmers and was published in the regional farmers' bulletins in the described catchment area. The questionnaire inquired about the saving of antibiotics during drying off, the criteria guiding the farmer's decision (cell count from monthly dairy herd improvement data (DHI), mastitis history, microbiological examination of quarter foremilk samples, California mastitis test), use of teat sealants and the type of dry off procedure (abrupt/gradual). RESULTS: A total of 103 questionnaires were evaluated, making the response rate ~1% for Hesse, ~3% for Saarland, and ~5% for Rhineland-Palatinate based on the number of included farms. Approximately 29% of the farmers dried off one out of four cows, 20% half, 23% three out of four and 13% all cows without using antibiotics. Eighty-nine farm managers based their decision on the somatic cell counts of DHI. Additional criteria influencing the decision were the mastitis history, results of the California Mastitis Test, or a combination of both. In 76 farms cows were dried off abruptly. In 79 farms teat sealers were used. CONCLUSIONS: Application of SDCT is established in most of the farms that participated in the survey, even though the proportion varied between farms. Legal requirements are not the only reason farmers need to increasingly deal with SDCT; sustainability programs of the dairies rely on selective drying off as well. Herd veterinarians should be supportive in implementing these measures to achieve good udder health while reducing the use of antimicrobials to a necessary minimum.

Influence of birth weight, sex and disinfection on the involution of umbilical structures in calves.

BACKGROUND: Umbilical disinfection for calves is often recommended, but scientific studies do not always show a positive effect on disease prevention. In addition, there is little to no metric data on umbilical involution or on factors influencing the size of umbilical structures. Therefore, the aim of this study was to investigate the physiological involution of umbilical structures and the effect of two different umbilical treatment methods. METHODS: After birth, calves' umbilical cord stumps were treated with either an iodine tincture (n = 43) or an antibiotic spray (n = 41) or were left untreated (n = 42). On days 0, 1, 3, 7 and 14 after birth, calves were examined and external umbilical structures were measured. In addition, on day 14 (±2 days), a sonographic examination of umbilical structures was conducted. RESULTS: The treatment applied had no significant effect on the size of umbilical structures during the first 14 days of life. However, the diameter of the umbilical stalk was larger in male calves and calves with a higher birth weight than in female calves and calves with a lower birth weight. LIMITATION: The study was carried out on only one farm, so the generalisability of the findings may be limited. CONCLUSION: Umbilical structures showed significant involution over time and were influenced by birth weight and sex.

Ultrasound characterization of mammary gland development in heifer calves fed at two different levels until weaning.

Ultrasound technologies allow for a non-invasive assessment of mammary gland (MG) development, the differentiation between the tissue types of the MG, and the evaluation of changes in its composition. This study aimed to work out a detailed description of the different stages of MG development that are visually discernible by ultrasonography for providing a template to classify the different structures. With this basis, the qualitative categorization of the developmental stage, as well as further quantitative assessments via pixel densities in the structures of interest, should be facilitated. Ultrasonic images were acquired from all four quarters of 37 German Holstein heifer calves fed either at a high feeding level of milk replacer (MR; 14% solids) at 10 L/day (1.4 kg MR/day; HI, n = 18) or at a restrictive low level of 5.7 L/day (0.8 kg MR/day; RES, n = 19) until linear weaning from week 13 to 14 of life. Ultrasound MG scans were performed first in week 3 of life, fortnightly from week 8-16, and in week 20 of life, in standing position, of each quarter, using a B-mode ultrasound device equipped with a linear probe (18 MHz). The developmental stages of the mammary gland parenchyma (PAR), visible in ultrasound images, obtained over 20 weeks of life, were categorized, described, and drawn by hand. On this basis, a template for classifying the visible categories of mammary PAR development and its surrounding tissue (SURR), and for measuring their pixel brightness was created thus providing an ultrasonographic atlas of the developing bovine MG, describing 11 categories. The ultrasound images were further classified by PAR structure, and pixel brightness was measured in PAR and SURR by using ImageJ Fiji. The difference in pixel brightness between PAR and SURR, the delta (Δ) pixel value was calculated. With increasing age, higher atlas categories of PAR developmental stages were shown. Pixel values, i.e. the brightness of PAR, its SURR, and Δ pixel value changed with age but were neither affected by the feeding group nor by a group × time interaction. With progressing PAR development, its pixel brightness increased from week 10 to 20 of life, i.e., the PAR became more hyperechoic since it spread and grew into its SURR. The atlas can serve as a template for the categorization and qualitative assessment of MG structures and for the quantitative assessment of PAR's development by measuring pixel brightness. With our study, we could show the structural development in PAR as well as in SURR in MG simultaneously in early life and confirm the spreading of PAR into its SURR by ultrasound scanning.

Inter- and Intra-Individual Variation in the Behavior of Feed Intake on Nutrient Availability in Early Lactating Dairy Cows.

Since energetic deficits in dairy cows can only be reduced at an animal level, the objective of the present study was to determine the extent of variation in intake behavior within and between animals during early lactation, to explore the magnitude of interactions between feed intake, intake behavior and nutrient digestibility, and to identify levers for maximizing feed intake at the individual animal level. Feeding behavior, intake and nutrient digestibility of 28 German Holstein dairy cows, fed TMR with 7.0 MJ NEL, were studied between the 2nd and 15th week after calving. Dry matter intake was assessed daily and nutrient digestibility weekly, with iNDF240 as an intrinsic marker. Results showed high intra- and inter-individual variation in intake behavior parameters with coefficients of variation (CV) up to 0.58 in meal frequency. Nutrient digestibility varied only slightly with CV values up to 0.10 in crude protein. Milk yield, meal frequency, feeding time, feeding rate and meal size had significant positive effects on DMI (p < 0.01). To achieve long-term improvements in feed intake, it is important to optimize feed intake and feeding behavior of individual animals by improving feeding conditions and develop technical tools to identify animals with insufficient feed intake.

Characteristics of the Oxidative Status in Dairy Calves Fed at Different Milk Replacer Levels and Weaned at 14 Weeks of Age.

A paradigm shift in the way of rearing heifer calves from restricted feeding and early weaning towards greater feed allowances and later weaning ages is ongoing. We aimed at characterizing the oxidative status in Holstein heifer calves fed with milk replacer (MR) at either a restrictive (RES) or a high (HIGH) level for 14 weeks. We compared two groups: HIGH (10 L MR/d, n = 18) and RES (5.7 L/d, n = 19) from day five until week 14 of life. In blood samples collected at birth, and then fortnightly from week 8-16, and in week 20, the antioxidative capacity measured as ferric reducing ability of plasma (FRAP), oxidative damage of lipids measured as thiobarbituric acid reactive substances (TBARS) and oxidative damage of proteins measured as advanced oxidation products of proteins (AOPP), free radicals measured as reactive oxidative metabolites (dROM), and the glutathione peroxidase (GSH-Px) activity, as well as leptin, adiponectin and haptoglobin were assessed. The time course of these variables during the first 20 weeks of life showed characteristic patterns; group differences were limited to adiponectin, AOPP, and FRAP. RES calves had lower growth rates, showed signs of hunger, but did not differ from HIGH in their intake of solid starter feed and in health status. This work characterizes the changes in oxidative status of dairy calves with increasing age and confirms the benefits of a high feeding plane with regard to welfare and development.

Association between alterations in plasma metabolome profiles and laminitis in intensively finished Holstein bulls in a randomized controlled study.

Metabolic consequences of an energy and protein rich diet can compromise metabolic health of cattle by promoting a pro-inflammatory phenotype. Laminitis is a common clinical sign, but affected metabolic pathways, underlying pathophysiology and causative relationships of a systemic pro-inflammatory phenotype are unclear. Therefore, the aim of this study was to elucidate changes in metabolome profiles of 20 months old Holstein bulls fed a high energy and protein diet and to identify novel metabolites and affected pathways, associated with diet-related laminitis. In a randomized controlled feeding trial using bulls fed a high energy and protein diet (HEP; metabolizable energy [ME] intake 169.0 ± 1.4 MJ/day; crude protein [CP] intake 2.3 ± 0.02 kg/day; calculated means ± SEM; n = 15) versus a low energy and protein diet (LEP; ME intake 92.9 ± 1.3 MJ/day; CP intake 1.0 ± 0.01 kg/day; n = 15), wide ranging effects of HEP diet on metabolism were demonstrated with a targeted metabolomics approach using the AbsoluteIDQ p180 kit (Biocrates Life Sciences). Multivariate statistics revealed that lower concentrations of phosphatidylcholines and sphingomyelins and higher concentrations of lyso-phosphatidylcholines, branched chain amino acids and aromatic amino acids were associated with an inflammatory state of diet-related laminitis in Holstein bulls fed a HEP diet. The latter two metabolites share similarities with changes in metabolism of obese humans, indicating a conserved pathophysiological role. The observed alterations in the metabolome provide further explanation on the underlying metabolic consequences of excessive dietary nutrient intake.

Sphingolipid Profiling Reveals Different Extent of Ceramide Accumulation in Bovine Retroperitoneal and Subcutaneous Adipose Tissues.

Sphingolipids are bioactive lipids that can modulate insulin sensitivity, cellular differentiation, and apoptosis in a tissue-specific manner. However, their comparative profiles in bovine retroperitoneal (RPAT) and subcutaneous adipose tissue (SCAT) are currently unknown. We aimed to characterize the sphingolipid profiles using a targeted lipidomics approach and to assess whether potentially related sphingolipid pathways are different between SCAT and RPAT. Holstein bulls (n = 6) were slaughtered, and SCAT and RPAT samples were collected for sphingolipid profiling. A total of 70 sphingolipid species were detected and quantified by UPLC-MS/MS in multiple reaction monitoring (MRM) mode, including ceramide (Cer), dihydroceramide (DHCer), sphingomyelin (SM), dihydrosphingomyelin (DHSM), ceramide-1-phosphate (C1P), sphingosine-1-phosphate (S1P), galactosylceramide (GalCer), glucosylceramide (GluCer), lactosylceramide (LacCer), sphinganine (DHSph), and sphingosine (Sph). Our results showed that sphingolipids of the de novo synthesis pathway, such as DHSph, DHCer, and Cer, were more concentrated in RPAT than in SCAT. Sphingolipids of the salvage pathway and the sphingomyelinase pathway, such as Sph, S1P, C1P, glycosphingolipid, and SM, were more concentrated in SCAT. Our results indicate that RPAT had a greater extent of ceramide accumulation, thereby increasing the concentration of further sphingolipid intermediates in the de novo synthesis pathway. This distinctive sphingolipid distribution pattern in RPAT and SCAT can potentially explain the tissue-specific activity in insulin sensitivity, proinflammation, and oxidative stress in RPAT and SCAT.

Changes of the erythrocyte phenotype and blood biochemistry in dairy calves during the first ten weeks of age.

BACKGROUND: Bovine erythrocytes undergo important changes in their morphology and chemical composition during the first weeks of age, which must be understood to accurately interpret hematology results in calves. The objectives of this prospective cohort study were to describe physiological changes of calf erythrocytes and to investigate mechanisms potentially causing these changes. METHODS: Blood samples from 30 clinically healthy dairy calves were obtained from birth to the tenth week of age in weekly intervals. Hematological and plasma biochemical parameters as well as the mineral electrolyte content of erythrocytes were determined and followed over time. The changes of parameters characterizing the erythrocyte phenotype over time were compared to the changes of plasma and erythrocyte biochemical parameters and possible associations were investigated using correlation and stepwise regression analyses. RESULTS: Although the erythrocyte mean corpuscular volume (MCV) declined from 43.6 ± 3.7 fL to 35.6 ± 3.2 fL between the first and seventh week, the red blood cell count (RBC) increased from 7.2 ± 1.1 × 1012/L to 9.3 ± 1.0 × 1012/L until the fifth week of age. The blood hemoglobin (Hb) concentration increased from 0.96 ± 0.16 g/L to 1.16 ± 0.11 g/L in the first three weeks of age and remained at this level until the end of the study. Changes in MCV were accompanied by a decline of the erythrocyte potassium content (KERY) from 91.9 ± 13.5 to 24.6 ± 7.2 mmol/L and a concomitant increase of the erythrocyte sodium content from 45.0 ± 32.0 to 102.7 ± 26.5 mmol/L. MCV was found to be associated with KERY, the primary determinant of the intra-erythrocyte osmotic pressure from the sixth week of age and with blood hemoglobin, the primary determinant of the intra-erythrocyte oncotic pressure from the eighth week of age, when KERY, blood Hb and MCV already had reached or approached normal levels of adult cattle. The plasma iron concentration was not found to be associated to any of the studied hematological parameters. CONCLUSION: A volume reduction of 20% in bovine neonatal erythrocytes is a physiological change occurring during the first weeks of age and is neither associated with sideropenia nor with anemia in healthy calves. The mechanism driving the observed erythrocyte volume change could not be identified. Results of the correlation and regression analyses indicate that changes in intra-erythrocyte osmotic or oncotic pressure are improbable underlying causes. Results reported here show that KERY is an unreliable indicator for the K homeostasis of the intracellular space in neonatal calves and that a decrease in MCV in early life per-se is an unreliable indicator for the development of microcytic anemia.

Understanding barriers to following advice: Evaluation of an advisory service from dairy farmers' perspectives.

In dairy herd health medicine, the success of consultation is sometimes limited as farmers do not always implement given advice. Therefore, the objective of this study was to evaluate a consultation performed during a study in the northwest of Germany and thereby to detect barriers that hinder farmers with long lasting problems in herd health from implementing veterinary advice. Six to twelve months after a non-recurring extensive herd health analysis and consultation, 38 farmers were interviewed by phone. Nearly all farmers were content with the farm visit, and 79% of the farmers stated that they had implemented at least some of the advice. This shows that farmers appreciated this service in general and were willing to follow advice. Even though the results cannot be generalized, factors that could be considered by advisors to improve the success of consultation were detected: Reasons as to why the advice was rejected were mostly related to farmers' expectations. Implementing only some of the advice was caused by a lack of time, self-discipline, money, and a lacking farm successor. Factors that pleased farmers were friendliness of the study veterinarians, in-depth examinations, handling of cows, good advice and how well organized the farm visit was. Factors that displeased the farmers were usually indicated only by one farmer each. Other factors influencing the success of consultation were the teamwork with the practising veterinarian, the self-evaluation of the farmers and the desire of the farmers for a single reason for the herd health problems.

Potential role of real-time PCR for detecting Mycobacterium avium subsp. paratuberculosis in chronically diseased milking cows: a case control study.

Paratuberculosis caused by Mycobacterium avium subsp. paratuberculosis (MAP) is economically important to dairy operations. In the present study, a real-time PCR kit (certified for use in Germany) was used to detect MAP in bovine fecal and milk samples within a case control study examining different factors for their association with chronic disease in Northern Germany.The aim of this study was to describe the suitability of the MAP real-time PCR kit to detect MAP in feces and milk of chronically diseased dairy cows. For MAP detection a total of 928 fecal, 922 composite foremilk, and 92 bulk milk samples obtained from 58 cases and 35 control dairy herds were investigated. The real-time PCR showed MAP positive results for 11 (18.96%) and 6 (17.14%) of the case and control herds, respectively. All bulk milk samples were MAP negative. The results of fecal and milk samples were moderately correlated (kappa = 0.27). These data indicate that real-time PCR results have diagnostic value for diagnosing MAP positive animals, and that fecal samples are more suitable than milk samples for assessing the reasons for chronic disease on dairy farms.

Occurrence of Clostridium botulinum neurotoxin in chronic disease of dairy cows.

Botulism caused by neurotoxins of Clostridium (C.) botulinum is a rare, but serious life-threatening disease in humans and animals. Botulism in livestock is usually caused by the oral uptake of C. botulinum neurotoxins (BoNT) via contaminated feed and is characterized by flaccid paralysis. In the recent past a new syndrome caused by BoNT in dairy cattle was postulated. It was supposed that C. botulinum is able to colonize the lower intestine and may subsequently produce neurotoxin. The continuous resorption of small amounts of these BoNT may then provoke the so called syndrome of "chronic" or "visceral" botulism involving unspecific clinical symptoms, reduced performance of dairy cows and massive animal losses in the affected herd. To test this hypothesis a case-control study was conducted involving 92 affected farms and 47 control farms located in Northern Germany. Fecal samples of 1388 animals were investigated for the presence of BoNT to verify the key requirement of the hypothesis of chronic botulism. BoNT was not detected in any of the fecal samples using the most sensitive standard method for BoNT detection, the mouse bioassay. Therefore, the existence of "chronic" or "visceral" botulism could not be proven.