Effects of an intensive experimental protocol on health, fertility, and production in transition dairy cows.

Animal experimentation is required to investigate complex physiological relationships and facilitates development of evidence-based knowledge. However, experimental protocols can interfere with the daily routine of the animals, result in stress and pain, and have adverse effects on health and production. The goal of this study was to investigate the effects of an intensive experimental protocol on health traits and production in transition dairy cows. Eighty experimental dairy Holstein cows (EXP group) underwent serial protocol-based clinical and ultrasonographic examinations, puncture of the jugular vein for blood collection or drug application, and liver biopsy samples, 14 d before until 42 d after parturition. Controls (CTR group) included 206 cows from the same herd, which fulfilled the same inclusion criteria and were kept under the same production management but were not handled for the purpose of this study. Procedure-related effects with a potentially negative effect on health and production were recorded. Furthermore, production, fertility and culling traits of the 2 groups (CTR, EXP) were compared. Most procedure-related adverse effects were associated with transcutaneous liver biopsies and included diffuse inflammation of the skin incision in 11.9% (42 of 320), abscessation of the skin or subcutis in 4.6% (11 of 240), and increased liver echogenicity of the biopsy site in 10.4% (27 of 240). The experimental procedures had a negative effect on milk yield at the start [days in milk (DIM) 5-50, difference: 2.3 kg, standard error (SE): 0.8 kg] and end of lactation (DIM 251-300, difference: 2.0 kg, SE: 1.0 kg; DIM 301-350, difference: 2.3 kg, SE: 1.2 kg) resulting in a lower 305-d milk yield in the EXP group than in the CTR group (difference: 472 kg, SE 214 kg). The milk fat % was higher in the EXP group than in the CTR group from 251 DIM onward (DIM 251-300, difference: 0.20%, SE: 0.09%; DIM 301-350, difference: 0.41%, SE: 0.17%). Also, the somatic cell score was higher in the EXP group than in the CTR group, during early (5-50 DIM, difference: 0.43, SE: 0.22) and from mid-lactation onward (DIM 151-200, difference: 0.43, SE: 0.2; DIM 201-250, difference: 0.49, SE: 0.22; DIM 251-300, difference: 0.55, SE: 0.25; DIM 301-350, difference: 0.61, SE: 0.28). Experimental procedures had no effect on first service conception rate and time to pregnancy, but had a positive effect on stillbirth rate with fewer stillbirths in the CTR group (0%) than in the EXP group (3.9%). Furthermore, experimental handling had no effect on time to culling or type of culling, whereby poor production was a more frequent reason noted for culling in the EXP group. Procedure-associated impairment of production in dairy cows is rarely reported and allows the estimation of the effects of such a study protocol on animal health and production. As a limitation for the interpretation of the results, the number of animals included and conduction in one single herd have to be considered.

Effects of a 6-wk intraduodenal supplementation with quercetin on energy metabolism and indicators of liver damage in periparturient dairy cows.

Periparturient dairy cows experience metabolic challenges that result in a negative energy balance (EB) and a range of postpartum health problems. To compensate for the negative EB, cows mobilize fatty acids from adipose tissues, which can lead to fatty liver disease, a periparturient metabolic disorder. Flavonoids, such as quercetin (Q), are polyphenolic substances found in all higher plants and have hepatoprotective potential and the ability to prevent or reduce lipid accumulation in the liver. In ruminants, few studies on the metabolic effects of Q are available, and thus this study was conducted to determine whether Q has beneficial effects on EB, lipid metabolism, and hepatoprotective effects in periparturient dairy cows. Quercetin was supplemented intraduodenally to circumvent Q degradation in the rumen. Cows (n=10) with duodenal fistulas were monitored for 7wk. Beginning 3wk before expected calving, 5 cows were treated with 100mg of quercetin dihydrate per kilogram of body weight daily in a 0.9% sodium chloride solution for a total period of 6wk, whereas the control cows received only the sodium chloride solution. The plasma flavonoid levels were higher in the Q-treated cows than in the control cows. A tendency for higher postpartum (pp) than antepartum (ap) plasma flavonoid levels was observed in the Q-treated cows than in the controls, which was potentially caused by a reduced capacity to metabolize Q. However, the metabolic status of the Q-treated cows did not differ from that of the control cows. The pp increases in plasma aspartate aminotransferase and glutamate dehydrogenase activities were less in the Q-treated cows than in the control cows. The Q had no effect on energy expenditures, but from ap to pp the cows had a slight decline in respiratory quotients. Irrespective of the treatment group, the oxidation of fat peaked after calving, suggesting that the increase occurred because of an increased supply of fatty acids from lipomobilization. In conclusion, supplementation with Q resulted in lower pp plasma aminotransferase and glutamate dehydrogenase, which indicated reduced liver damage. However, the direct effects of Q on the liver and the implications for animal performance remain to be investigated.

Zearalenone (ZEN) metabolism and residue concentrations in physiological specimens of dairy cows exposed long-term to ZEN-contaminated diets differing in concentrate feed proportions.

A long-term feeding experiment with dairy cows was performed to investigate the effects of feeding a Fusarium toxin contaminated (FUS) and a background-contaminated control (CON) ration with a mean concentrate feed proportion of 50% during the first 11 weeks after parturition (Groups FUS-50, CON-50, Period 1), and with concentrate feed proportions of 30% or 60% during the remaining 17 weeks (Groups CON-30, CON-60, FUS-30 and FUS-60, Period 2), on zearalenone (ZEN) residue levels in blood serum, milk, urine and bile. ZEN, α-zearalenol (α-ZEL) and ß-zearalenol (ß-ZEL), zearalanone (ZAL), α-zearalanol (α-ZAL) and ß-zearalanol (ß-ZAL) were determined by HPLC with fluorescence detection. The ZEN concentrations of the rations fed to Groups CON-50, FUS-50 (Period 1), CON-30, CON-60, FUS-30 and FUS-60 (Period 2) amounted to 53.1, 112.7, 35.0, 24.4, 73.8 and 72.5 µg/kg dry matter, respectively. The concentrations of ZEN, α-ZEL, ß-ZEL, ZAN, α-ZAL and ß-ZAL in serum, urine and milk were lower than 1, 1, 4, 100, 50 and 200 ng/g, respectively, while ZEN, α-ZEL and ß-ZEL were detected in bile. Their levels changed with oral ZEN exposure in the course of the experiment and in a similar direction with concentrate feed proportion (Period 2 only). Thus the proportions of the individual ß-ZEL, α-ZEL and ZEN concentrations of their sum varied only in narrow ranges of 68-76%, 6-13% and 12-20%, respectively. Interestingly, the bile concentrations of ß-ZEL, α-ZEL and ZEN of Groups CON-60 and FUS-60 amounted to only approximately 50%, 45% and 62%, respectively, of those of Groups CON-30 and FUS-30 despite a similar or even lower ZEN exposure. The results indicate that conversion of ZEN to its detectable metabolites was not changed by different dietary concentrate feed proportions while their absolute levels were decreased. These findings might suggest concentrate feed proportion-dependent and rumen fermentation-mediated alterations in ZEN/metabolite degradation, and/or liver associated alterations in bile formation and turnover.

A Sustainable Translational Sheep Model for Planned Cesarean Delivery of Contraction-Free Ewes.

We evaluated whether the sheep constitutes a useful translational model to evaluate anatomical and surgical aspects of cesarean delivery (CD) from a human medical perspective with the aim of both maternal and neonatal well-being. Our hypothesis was that CD in contraction-free ewes is not associated with major complications. Primary endpoint was the transferability of anatomical conditions and surgical techniques of CD from the ewe to the human. Secondary endpoints were maternal and fetal survival, occurrence of retained fetal membranes, metritis, mastitis, or wound infections. Forty-eight Merino ewes were delivered by CD after 95% gestation (142-144 days). Both ewes and newborn lambs were cared for intensively after the delivery. Ovine uterine anatomy during CD appeared slightly different but comparable to the human uterus. Uterine incisions were mostly performed in the uterine horns, not in the uterine corpus. The ovine uterine wall is thinner than in humans. All ewes survived without any major complications. Seventy-seven (88.5%) out of 87 live-born lambs survived without any complications. The contraction-free ewe constitutes an appropriate and safe model to evaluate anatomical and surgical aspects of CD from a human medical perspective. We present a step-by-step manual for successfully planned cesarean delivery for sheep including the perioperative management illustrated with photographs and a five-minute video. With adequate planning and a reasonable number of staff, it is possible to safeguard both maternal and neonatal survival. This sustainable translational medicine model offers additional potential for the offspring to be used for further research studies (e.g., transgenerational inheritance research).

A method for estimating the outer exposure of dairy cows to deoxynivalenol (DON) and zearalenone (ZEN) as a precondition for risk assessment based on inner exposure with toxin residue levels in blood and urine as indicators.

DON and ZEN residues in the blood and urine of dairy cows can be used to predict the outer exposure to DON and ZEN expressed per kilogram diet for a risk evaluation based on comparisons to critical dietary concentrations. This method was used to evaluate the exposure of dairy cows from 12 farms located in Brandenburg, Germany, fed rations with unknown DON and ZEN concentrations (N = 244). The corresponding diet concentrations predicted by different methods from analyzed blood and urine samples varied significantly amongst farms from 0 to 1.6 mg/kg for DON and 0 to 3.0 mg/kg for ZEN at a reference dry matter content of 88% but independently of lactational state (post-partum vs. early lactation). This significant variation was noticed below the critical dietary DON concentration of 5 mg/kg, while the ZEN concentration in one farm exceeded the critical ZEN level of 0.5 mg/kg markedly. Predicted DON concentrations of rations increased with the proportion of maize silage, while the high ZEN concentration found in one farm was most likely related to a higher proportion of sugar beet pulp supposedly highly contaminated by ZEN. Exceeding the critical dietary ZEN concentration and significant variations in DON contents below the critical level was not related to performance, reproductive performance, and health-related traits of cows. For a more consistent evaluation of possible associations between the inner exposure of cows to DON and ZEN, more frequent longitudinal observations of both mycotoxin residue levels and performance and health traits are required.

Comparative cryopreservation of bovine and porcine primary hepatocytes.

The isolation of primary hepatocytes from liver tissue of farm animals yields a very high number of cells, and a part of them can be stored by cryopreservation for future experiments. As no experience exists with the cryopreservation of hepatocytes from cattle, our study aimed at the cryopreservation of bovine hepatocytes by use of different protocols compared with the cryopreservation of hepatocytes from pig. We tested different freezing media (William's Medium E vs. University of Wisconsin solution), cryoprotectants (dimethyl sulfoxide with vs. without trehalose as additional additive), freezing systems (standard freezing container vs. controlled-rate freezer) and freezing times (4 vs. 28 d). These tests identified a general influence of species and freezing systems, whereas the influence of freezing media, trehalose additive and freezing time was less or not obvious. In this regard, we determined a mean recovery of 30% of bovine hepatocytes and 55% of porcine hepatocytes cryopreserved in a controlled-rate freezer, whereas the rates were about 10% less when hepatocytes were frozen in a standard freezing container. In accordance with this observation, the cultivation of cryopreserved hepatocytes from cattle was less effective than that of porcine hepatocytes. Hepatocytes from cattle can be successfully cryopreserved and partially cultured after cryopreservation but with lower percentage than porcine hepatocytes.

Case report: Teat stenosis in a suckler cow.

Bovine veterinarians are regularly confronted with teat lesions in cows. The number of studies on the diagnosis and treatment of teat lesions as well as the exchange of practical experience among clinicians are extensive in dairy cows compared with suckler cows. The aim of this case report was to describe the successful treatment of teat stenosis in a suckler cow and discuss possible challenges. A four-year-old Simmental cow, in her third lactation and 4 days in milk, was referred to our clinic along with her calf because of teat stenosis in the front left quarter. The owner had repeatedly used a rigid teat cannula in an attempt to relieve the stenosis during the previous lactation. However, the cow had refused to allow the current calf to suckle the affected teat and resisted attempts by the owner to cannulate the teat. The results of clinical examination, ultrasonography, and milk sampling showed stenosis of the proximal, middle, and distal parts of the front left teat cistern, accompanied by thelitis and cisternitis and mild chronic clinical mastitis. Based on published recommendations, treatment of the thelitis, cisternitis, and mastitis was initiated before resolution of the stenosis surgically. The first week of treatment included the administration of an intramammary product containing cefapirin and prednisolone, a systemic non-steroidal anti-inflammatory drug, a wax teat-boogie, and bandaging of the teat. Thereafter, the treatment was reduced to insertion of a wax-teat boogie and bandaging. Conservative treatment resulted in resolution of the mastitis, cisternitis, and stenosis in the proximal and middle parts of the teat, which had most likely been caused by repeated cannulation of the teat by the owner. Lateral theloscopy was then used to remove the distal stenosis, which was the primary lesion. Healing of the surgical wound and resolution of the swelling occurred several days postoperatively, and the calf's first attempt to suckle the teat was successful. The cow and calf were discharged from the clinic 2 weeks after surgery. A follow-up visit 4.5 months after surgery revealed that the calf was still nursing the teat and the operated quarter was producing a normal amount of milk.

Evaluation of multifactorial digestive disorders in a dairy herd at different stages of lactation.

The manager of a large dairy herd (total confinement, total mixed ration feeding, 10130 kg milk sold per cow and year) requested a workup of a digestive disorder problem that had been ongoing for several years. The cows were at all stages of lactation, and the incidence proportion (events/100 cows at risk) of digestive disorders was approximately 5 to 10%. The clinical picture included an abnormal demeanor of varying severity and signs that differed among the cows. The latter included decreased milk production and physical activity, low body condition score, abnormal rumen motility and stratification, small intestinal dilatation, diarrhea, undigested fiber particles in the feces, fever and abdominal pain.The following approach was used to investigate the digestive disorder problem:1. The herd was assessed for production levels, housing requirements, feeding protocols and animal health management. The latter comprised evaluation of different animal scores, metabolic profile analysis, diagnosis of disease, culling records and slaughter data. The results revealed risk factors concerning the feeding and animal health monitoring, (e. g. in dry matter intake and silage quality management, disease detection and diagnosis in fresh cows). The assessment also identified a high occurrence of digestive disorders of unknown origin.2. Fifteen cows that represented the ongoing digestive disorder problem were selected to undergo clinical examination, hematological analysis, urinalysis, and ultrasonography of the ventral abdomen. The clinical examinations revealed different digestive disorders, which were mainly inflammatory in nature, in all the cows. Eight cows had localized reticuloperitonitis and 13 had left displaced abomasum with different degrees of displacement and adhesions between the abomasum and reticulum attributable to reticuloperitonitis.Our results revealed a multifactorial problem caused by several risk factors relating to animal health and feeding management protocols that resulted in different types of digestive disorders. The wires from damaged tires used to hold the pit silo tarps in place were identified as a possible traumatic cause of the reticuloperitonitis. Treatment, prevention and follow-up of the different conditions were discussed.

[Treatment, clinical course and outcome of tibial neuropathy in German Holstein cows]. / Behandlung, Verlauf und Therapieerfolg der Neuropathie des Nervus tibialis bei Deutsch-Holstein Kühen.

OBJECTIVE: As a follow-up to a previous study on the incidence, history and clinical findings of tibial neuropathy (TN), the present work aimed at describing the treatment and prognosis of this disease. MATERIALS AND METHODS: Of 88 German Holstein dairy cows with unilateral (UTN, n = 71) or bilateral (BTN, n = 17) TN, 68 (56 UTN, 12 BTN) with a complete data set were analyzed. They were retrospectively assigned to one of four groups: no treatment - spontaneous healing within 48 h (Spontaneous, 5 UTN), no bandage (0Cast, 8 UTN, 3 BTN) or treatment with anti-inflammatory drugs and support bandage (StV, 3 UTN) or fiberglass cast (Cast, 40 UTN, 9 BTN). Treated cows were re-examined five times (14, 21, 28, 42 and 56 days after the first presentation). The plasma activity of creatine kinase was measured at the last re-examination in 29 cows similar to measurement at day 0. RESULTS: The observed overall success rate of treatment of cows with UTN was considerably higher compared with untreated cows (Cast 98 % and StV 100 % vs. 0Cast 62 %). By comparison, the observed difference between treated and untreated cows with BTN was not so clear (78 % vs. 67 %). Recovering cows exhibited a calculated longer median survival time than cows that did not recover (545 d vs. 100 d). Plasma creatine kinase activities were increased initially and returned within the reference interval (434 U/l and 152 U/L) following treatment. CONCLUSIONS: Cows with ETN have an excellent prognosis provided that treatment with anti-inflammatory drugs and stabilizing bandage is administered. In cows with BTN, the prognosis depended on the type and degree of the primary injury. Loss of skin sensitivity indicated a poor prognosis. From an economic standpoint, treatment of TN is indicated provided that the prognosis is good. In cows that had healed clinically, the average survival time extended into the following lactation. CLINICAL RELEVANCE: This study highlights the advantages of a support bandage for the treatment of cows with TN. Compared with other peripheral neuropathies, muscle damage appears to be of particular importance in TN.

Development of an electronic interface for transfer of antimicrobial administration data in dairy farms.

Surveillance of antimicrobial administration in livestock production is an important factor in global policies to reduce spreading of antimicrobial resistance. In recent years, many studies have been carried out concerning the usage of antimicrobials in animal production and in some countries recording of antimicrobial quantities dispensed to famers is mandatory. On cattle farms, antimicrobial treatments are recorded for fattening calves under 8 months of age and for fattening cattle older than 8 months in Germany and treatment frequencies are then calculated. However, with the entry into force of Regulation (EU) 2019/6 on 01/28/2022, antimicrobial monitoring will gradually be extended to all animal species and age groups. Therefore, an effective, fast and accurate transfer of data on the use of antimicrobials, especially in the field of livestock farming, into corresponding databases is required to determine the treatment frequencies for the individual animal species or types of use. For this purpose, an electronic interface was programmed to transfer the data on antimicrobial use in dairy cattle farms from a herd management software program directly into a database. To test the practicability and effectiveness of this interface, 10 dairy cattle farms from Saxony were initially selected. Based on an in-depth analysis of the treatment frequencies of antimicrobial administration of 7 different age groups of animals after a two-year observation period, the functionality of the electronic interface could be established. The greatest potential for reduction of antimicrobials is in newborn calves, as they represent the age group with the highest treatment frequency.

Qualitative and Quantitative Changes in Total Lipid Concentration and Lipid Fractions in Liver Tissue of Periparturient German Holstein Dairy Cows of Two Age Groups.

Fatty liver syndrome (FLS) is a common disease in high-producing dairy cows. Studies in humans suggest that the different hepatic lipid fractions play a role in this context. In dairy cows, little is known about the composition of fat stored in the liver, its periparturient dynamics, and the effect of cows' age. Therefore, our goal was to generate primary data in healthy cows to serve as reference values for future studies. Eight healthy German Holstein cows (2nd lactation, n = 3; ≥3rd lactation, n = 5) were examined 14 d antepartum and 7, 28, and 42 d postpartum. The examinations included clinical assessment, liver biopsy, blood sampling, and recording of milk yield. Total lipids (TL) in liver tissue were measured gravimetrically. The TL were separated into lipid fractions (triacylglycerol, TAG; phospholipids, PL; non-esterified fatty acids, NEFA; and cholesterol esters) using thin-layer chromatography, followed by gas chromatography for fatty acid determination. Concentrations of NEFA, ß-hydroxybutyrate, and cholesterol were analyzed in blood. Concentrations of TL, TAG, NEFA, and cholesterol esters in liver tissue and NEFA in blood increased in the periparturient period. The older cows had higher hepatic TL, TAG, and PL concentrations, higher relative hepatic concentrations of TAG in TL, higher NEFA concentrations in blood, a greater decrease in body condition, and higher milk yields between d 9 and 40 than the younger cows. We proposed that due to higher milk yield, older cows mobilized and deposited more fat in the liver, and the increase in hepatic TAG concentration was longer-lasting than in younger cows. Higher levels of structural lipids (PL) in older cows could be explained by higher demand for storage of TAG and cholesterol esters in lipid droplets or for the export of TAG via very-low-density lipoproteins. Results show that hepatic fat storage is a reversible process and does not necessarily cause clinical disease. Nevertheless, older cows have a more sustained and greater increase in hepatic TAG concentration, which may explain their increased risk of FLS. The results are limited in their extrapolation due to the small sample size and thereby possible selection bias but present a valuable basis for future studies.

Radiation doses from low-dose CT scans in SPECT/CT and PET/CT examinations: A survey in Germany.

AIM: Recently, dose reference levels (DRLs) have been defined in Germany for auxiliary low-dose CT scans in hybrid SPECT/CT and PET/CT examinations, based on data from 2016/17. Here, another survey from 2020 was evaluated and compared with the new DRLs as well as with similar surveys from foreign countries. METHODS: The survey, which had already been conducted in the Nordic countries, queried for various examinations including the following values: patient weight and height, volume CT dose index (CTDIvol), dose length product (DLP). For each examination, statistical parameters such as the third quartile (Q3) were determined from all submitted CTDIvol and DLP values. Additionally, for examinations comprising datasets from at least 10 systems, the third quartile (Q3-Med) of the respective median values of each system was calculated. Q3 and Q3-Med were compared with the newly published DRLs from Germany and values from similar studies from other countries. RESULTS: Data from 15 SPECT/CT and 13 PET/CT systems from 15 nuclear medicine departments were collected. For the following examinations datasets from more than 10 systems were submitted: SPECT lung VQ, SPECT bone, SPECT&PET cardiac, PET brain, PET oncology. Especially for examinations of the thorax and heart, the new DRLs are very strict compared to this study. The CTDIvol values for examinations of the head were lower in this study than the DRLs prescribe now. CONCLUSIONS: For certain examination types, there is a need for dose optimization at some clinics and devices in order to take into account the new DRLs in Germany in the future.

[Incidence, history and clinical findings of tibial neuropathy in German Holstein cows]. / Vorkommen, Vorbericht und klinisches Bild der Neuropathie des Nervus tibialis bei Deutsch-Holstein-Kühen.

OBJECTIVE: The aim of this study was to document the occurrence, history, and clinical findings of tibial neuropathy in German Holstein dairy cows in order to yield information regarding the importance and etiology of this disorder in dairy herds in Central Germany. MATERIALS AND METHODS: This prospective study was undertaken between January 2013 and October 2017 and included 88 German Holstein dairy cows with unilateral (UTN, n = 71) or bilateral (n = 17, BTN) tibial neuropathy. Data were collected from the history as well as the clinical and laboratory examinations. Three grades of paresis were defined. RESULTS: The percentage of cows with tibial neuropathy among all German Holstein dairy cows presented with disorders of the locomotor system in the study period amounted to 2.2 %. UTN was seen predominantly following dystocia or as a result of prolonged lateral recumbency in consequence to a primary disease. The majority of cases of BTN occurred in association with the resumption of cyclicity following calving. These were accompanied by clinical signs of a cauda equina syndrome. Increased plasma activities of creatine kinase (UTN 98 %; BTN 100 %) and aspartate aminotransferase (UTN 89 %; BTN 100 %) were observed in nearly all affected cows. CONCLUSIONS: Tibial neuropathy is a regularly occurring disorder in dairy herds in Central Germany. The condition results in pain and locomotor impairment and therefore constitutes a welfare concern. UTN and BTN have different causes, and the history combined with clinical signs (severity of paresis) provides etiological information. Increased enzymatic activities suggest a muscular involvement in the disease process.

Multidrug-resistant enterobacteria in newborn dairy calves in Germany.

We studied the prevalence of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in dairy calves as part of a routine health check protocol. In addition, data regarding antimicrobial use (AMU), farm hygiene, and farm management were collected in order to identify possible risks for ESBL occurrence. Ten farms participated in the study with a median of 781 milking cows (319-1701). All calves investigated were younger than two weeks with an average age of 6.8 (±3.9) days. The farms were visited and samples were collected twice at an interval of 7-11 months. Faecal samples diluted 1:10, were plated onto BrillianceTM ESBL agar in duplicates. After 24 hours at 37°C, colonies were counted and total colony forming units (cfu)/ml calculated. Bacteria species were identified biochemically. ESBL-production was phenotypically confirmed using the MICRONAUT-S ß-Lactamases system. Additionally, antimicrobial susceptibility was tested using VITEK® 2 technology. Phylotyping of E. coli isolates and screening for bla genes was performed by PCR. ESBL-producing enterobacteria were detected on all farms and 96.5% of calves investigated shed ESBL-positive bacteria. Of all ESBL-producing isolates, the majority were E. coli (92.9%), followed by Enterobacter cloacae (5.1%) and Klebsiella pneumoniae subsp. pneumoniae (2.0%). The majority of E. coli isolates was clearly assigned to phylogroup C (25.0%), followed by phylogroups A (15.2%) and E (14.1%). CTX-M group 1 was most frequently detected (80.4%). E. cloacae contained blaCTX-M and blaTEM or blaSHV. K. pneumoniae harboured blaSHV only. Besides resistance to penicillins and cephalosporins, the majority of isolates was also resistant to one or more antibiotic classes, with a high proportion being resistant against fluoroqinolones. 52.5% of isolates were further characterised as threefold multidrug resistant gram-negative bacteria (3MDR-GNB) according to the German Commission for Hospital Hygiene and Infection Prevention. None of the isolates were 4MDR-GNB, i.e. none revealed carbapenem-resistance. Penicillins were the most frequently administered antibiotics to calves on most farms and were the predominant substance class at herd level on all farms. Overall, the number of calves treated prior to sampling was rather low (11.7%). Analyses of data regarding the farm management identified weaknesses in biosecurity and cleaning and disinfection. Besides beta-lactam antibiotics being the most commonly used antibiotics no other risk factors could be identified. In summary, the prevalence of ESBL-carriers in dairy calves was exceptionally high and should be motivation to develop strategies for the reduction of multidrug-resistant bacteria in farm animals.

Malnutrition as the cause of recumbency in suckler cows associated with Trypanosoma theileri infection.

BACKGROUND: Recumbent cows are a diagnostic challenge because of a wide range of differential diagnoses, which include trauma, neurological and metabolic disorders, malnutrition and mineral deficiencies. This case report describes recumbent suckler cows that presented as a herd problem. In addition to weakness due to inanition, Cu and Se deficiencies were considered as possible aetiologies of the recumbency. Furthermore, Trypanosoma (T.) theileri, a blood parasite of unknown importance in Germany, was detected in the blood of some cows. CASE PRESENTATION: Three recumbent cows were referred to the Clinic for Ruminants and Swine, Faculty of Veterinary Medicine of the University of Leipzig. They were unable to rise and had low body condition scores and rough hair coats. Haematological and serum biochemical analyses showed neutrophilia, electrolyte imbalances, increased activities of muscle and liver enzymes and decreased concentrations of trace elements, especially Copper (Cu) and Selenium (Se). T. theileri was detected in a routine blood smear from one cow. The cows did not respond to an intensive care protocol, which included intravenous fluids and electrolytes, mineral substitution, non-steroidal anti-inflammatories and antibiotics, and were therefore euthanized or died. Postmortem examination showed cachexia, subcutaneous and scleral oedema and muscular dystrophy, especially in the hind limbs. Follow-up examination of the herd of origin produced similar findings including the detection of T. theileri in a large proportion of the herd. Ration analysis revealed considerable undersupply of several nutrients. CONCLUSIONS: Based on all findings, an aetiological diagnosis of trace mineral and nutrient deficiency with possible involvement of T. theileri was made.

Case reports of squamous cell carcinoma in cows.

The aim was to investigate the frequency of squamous cell carcinoma and the outcome after surgical treatment in cows admitted to the clinic between 2012 and 2018. The study included six cows with squamous cell carcinoma (confirmed in 5 cattle and suspected in 1). All cattle underwent clinical examination and additional testing to confirm the diagnosis. Breed, age, gender, localization of neoplasm and treatment were recorded. In a telephonequ estionnaire owners were asked about survival, culling, integration into the herd after discharge from the clinic, wound healing and neoplasm recurrence. Five ocular squamous cell carcinomas were diagnosed during the study period. Another neoplasm on the dorsolateral thoracic wall of a cow could not be diagnosed with certainty; however, it had characteristics of squamous cell carcinoma. Two cows with ocular squamous cell carcinomawere euthanised.  The prognosis and outcome of squamous cell carcinoma depend on early detection of clinical signs, the presence of metastases and the timing of tumour removal. Surgical removal of a squamous cell carcinoma at or near the eye or at other locations of the body appears to have a good prognosis.

[Validation of a cow-side blood test for beta-hydroxybutyrate concentration in dairy cows]. / Überprüfung eines Schnelltestsystems zur Erfassung der Betahydroxybutyrat-Konzentration im Blut von Milchkühen.

OBJECTIVE: Clinical ketosis is common during the dairy cows' transition period and is responsible for considerable economic loss. Early identification of cows with subclinical ketosis is the first step for maintaining the health and productivity of dairy cows. The goals of the study were two-fold: The first was to examine the usefulness of a mobile test device as a cow-side test; and the second was to compare BHB concentrations measured by the ketometer using capillary blood and blood collected from the coccygeal vessels with values determined by a reference method in the laboratory using jugular blood. MATERIAL AND METHODS: Blood samples were collected from a jugular vein or the coccygeal vessels in 81 dairy cows at 7 time points (14 and 7 days pre-partum and 7, 14, 21, 28, and 42 days post-partum) for kinetic enzymatic measurement of BHB concentration in the laboratory. Blood samples were concurrently collected from the coccygeal vessels or by pricking the vulvar lip at the transition of the skin to the mucosa (capillary blood) to determine BHB concentration using the WellionVet BELUA ketometer (MED TRUST GmbH, Marz, Austria). RESULTS: Initial errors in operating the ketometer were quickly eliminated with experience. BHB concentrations of jugular blood measured in the laboratory were 0.07 mmol/l lower than those measured in coccygeal blood. The mean BHB concentration measured in coccygeal and capillary blood using the WellionVet BELUA ketometer did not significantly differ but were 0.13 and 0.12 mmol/l respectively, lower than the mean jugular vein concentrations measured in the laboratory. CONCLUSION: The WellionVet BELUA ketometer is useful for determination of BHB concentration in cows provided that the manufacturer's specifications are followed. Capillary blood is best collected at the transition from the vulvar skin to its mucosa. The device generates rapid results that correlate well with BHB concentrations determined in the laboratory and with the results obtained from different blood collection sites. It is ideally suited for monitoring dairy cows for subclinical ketosis using capillary or coccygeal blood.

[Bovine teat lesions - Retrospective data evaluation of 116 German Holstein cows]. / Zitzenverletzungen ­ retrospektive Datenauswertung von 116 Deutsch-Holstein-Kühen.

OBJECTIVE: The management and treatment of teat injuries represent major challenges in modern dairy farming. The injury per se, its treatment and the handling of the cow during the healing process interfere with the principal purpose of the cow, which is the production of saleable milk. The study presents treatment and post-operative management based on the evaluation of a large data set. MATERIALS AND METHODS: The data from patients of the Clinic of Ruminants and Swine of the University Leipzig from May 2012 to July 2017 were analysed in a retrospective study. The location and type of injury, initial and post-operative medication, and treatment and outcome were analyzed. RESULTS: In total, 120 teat injuries in 116 cows were treated. The teat canal was the most common location of injuries (n = 85; 70.8 %), and closed injuries (without an external wound) were the most frequent type (n = 73; 60.8 %). Of all teat canal injuries, 65 (54.2 %) were internal and did not involve the skin. Most of these injuries were treated under theloscopic guidance directly after initial examination irrespective of concomitant disorders, such as thelitis (except severe acute), cysternitis and mastitis. The majority of treated teats (n = 98; 87.4 %) had functional healing and could be milked, 8 (7.2 %) of treated teats were amputated and in the remaining functional healing was achieved after a second surgery. According to an owner survey at least 700 days post-treatment, 23 (21.5 %) of the surgically treated cows and 50 % (n = 2) of the non-surgically treated cows were slaughtered in consequence of complications associated with the treated teat or the corresponding quarter. CONCLUSION AND CLINICAL RELEVANCE: Teat injuries are commonly seen in dairy practice. A successful outcome is obtainable when surgical or medical treatment is administered promptly and consequently in addition to an appropriate post-operative management and monitoring.

Uptake and fecal excretion of Coxiella burnetii by Ixodes ricinus and Dermacentor marginatus ticks.

BACKGROUND: The bacterium Coxiella burnetii is the etiological agent of Q fever and is mainly transmitted via inhalation of infectious aerosols. DNA of C. burnetii is frequently detected in ticks, but the role of ticks as vectors in the epidemiology of this agent is still controversial. In this study, Ixodes ricinus and Dermacentor marginatus adults as well as I. ricinus nymphs were fed on blood spiked with C. burnetii in order to study the fate of the bacterium within putative tick vectors. METHODS: Blood-feeding experiments were performed in vitro in silicone-membrane based feeding units. The uptake, fecal excretion and transstadial transmission of C. burnetii was examined by quantitative real-time PCR as well as cultivation of feces and crushed tick filtrates in L-929 mouse fibroblast cells and cell-free culture medium. RESULTS: Ticks successfully fed in the feeding system with engorgement rates ranging from 29% (D. marginatus) to 64% (I. ricinus adults). Coxiella burnetii DNA was detected in the feces of both tick species during and after feeding on blood containing 105 or 106 genomic equivalents per ml blood (GE/ml), but not when fed on blood containing only 104 GE/ml. Isolation and cultivation demonstrated the infectivity of C. burnetii in shed feces. In 25% of the I. ricinus nymphs feeding on inoculated blood, a transstadial transmission to the adult stage was detected. Females that molted from nymphs fed on inoculated blood excreted C. burnetii of up to 106 genomic equivalents per mg of feces. CONCLUSIONS: These findings show that transstadial transmission of C. burnetii occurs in I. ricinus and confirm that I. ricinus is a potential vector for Q fever. Transmission from both tick species might occur by inhalation of feces containing high amounts of viable C. burnetii rather than via tick bites.

Field Trial Vaccination against Cowpox in Two Alpaca Herds.

In Europe, cowpox virus (CPXV) infection in South American camelids occurs as a so-called spill-over infection. Although infected animals generally have a mild form of the disease and survive, cases of fatal generalised CPXV infection have also been described. Prevention by prophylactic vaccination is the only way to protect animals from disease. In the present study, modified vaccinia virus Ankara (MVA) vaccine, which has been successfully used in many animal species, was used in a prime-boost vaccination regimen in two alpaca herds with a history of CPXV infection. The focus of the study was the prevention of further clinical cases, and to determine the safety and immunogenicity of the MVA vaccine in alpacas. The MVA vaccine was well tolerated and safe in the 94 animals vaccinated. An indirect immunofluorescence assay (IFA) using MVA as an antigen showed that the seroprevalence of antibody after booster vaccination was 81.3% in herd I and 91.7% in herd II. Detectable antibody titres declined to 15.6% in herd I and 45.8% in herd II over a 12-month period after booster vaccination. Animals could be divided into four groups based on individual antibody titres determined over one year: Group 1 consisted of 19.3% of animals that were seropositive until the end of the trial period; Group 2 consisted of 58.0% of animals that were seropositive after booster vaccination, but seronegative one year later; Group 3 consisted of 14.7% of animals that were not seropositive at any time point; and Group 4 consisted of 7.9% of animals that were seropositive after initial immunisation, seronegative six months later, but seropositive or intermediate in IFA one year after immunisation, likely because of natural exposure. In new-born crias born to MVA-vaccinated mares, specific maternal antibodies were detected in 50.0% of animals up to 14 weeks of age. Our results confirm that MVA vaccination is a feasible tool for the prevention of CPXV disease in alpacas. Long-term studies are needed to verify future vaccination regimen in CPXV affected herds.