Exposure of small ruminants to the Schmallenberg arbovirus in Germany from 2017 to 2018 - animal-specific and flock-management-related risk factors.

The Schmallenberg virus (SBV), an emerging Orthobunyavirus of mainly ruminant hosts, caused a substantial epidemic in European ruminant populations between 2011 and 2013. The pathogen is transmitted by arthropod vectors (Culicoides spp.) and can cause reproductive disorders and severe malformations of the offspring or stillbirth. The present study aimed to assess SBV seroprevalence among German sheep and goats a few years after the first virus detection in the country (November 2011). In addition, an extensive risk factor analysis including host-specific and husbandry-related factors was implemented. Seroprevalence was determined by examining serum samples from 2759 sheep and 446 goats out of a total of 70 flocks across five German federal states. The samples were withdrawn in the period between 2017 and 2018. Using a commercial competitive ELISA, antibodies against SBV were detected in all 70 investigated flocks. A percentage of 60.1 % (1657/2759) of the sheep and 40.4 % (180/446) of the goat sera contained SBV antibodies. Generalized linear mixed modeling revealed significant effects of host species (sheep > goats), age (old > young) and sex (female > male) on SBV seroprevalence. For both species, also the farming purpose, and for goats, ectoparasite treatment and the presence of cattle on the farm played a role in terms of risk for SBV exposure. The observations from this study still emphasize a wide distribution of the pathogen in Germany. Nevertheless, the observed seroprevalence might not be sufficient to achieve effective herd immunity. Pinpointing risk factors identified susceptible populations for targeted vaccination programs to reduce potential animal losses caused by SBV.

First Evidence of Leishmania infantum Antibodies in Sheep (Ovis aries) from Southern Germany.

In Europe, Leishmania infantum is the most prevalent Leishmania species, and this protozoan is transmitted by phlebotomine sandflies. A recent publication has shown that sheep harbor L. infantum antibodies. This raises questions about the epidemiological role of small ruminants. Therefore, sera from small ruminants located in two southern German federal states, Baden-Wuerttemberg (BW) and Bavaria (BAV), were analyzed with an ELISA to determine the presence of L. infantum antibodies. The species, sex and age (gimmer vs. ewe) were recorded, and a univariate analysis was conducted to determine possible associations. In total, seven sheep flocks (274 sheep/10 goats) from BW and seven sheep flocks (277 sheep/78 goats) from BAV were examined. In BW, four sheep from three flocks tested positive for L. infantum antibodies. In BAV, the same number of positive sheep were detected but in four flocks. The total seropositivity rate in sheep was 1.45%. All goats tested negative. No significant association (p > 0.05) was detected between Leishmania seropositivity and the variables evaluated. Our study reveals the exposure of sheep to L. infantum in a non-endemic area. Further investigation is needed to determine whether sheep can be used as sentinels to identify new phlebotomine habitats and Leishmania risk areas.

[Choanal atresia in an alpaca cria (Vicugna pacos)]. / Choanenatresie bei einem Alpakacria (Vicugna pacos).

After birth, an alpaca cria was noticed to exhibit weakness and respiratory distress, particularly when attempting to suckle milk from the dam's udder. Clinical findings indicated the presence of bilateral choanal atresia and the animal was subsequently euthanised. A computed tomography scan and a pathological examination were performed to describe the malformation in detail. Choanal atresia is a common malformation in neonatal South American camelids, surgical treatment is not recommended. This case report provides an overview of the clinical findings as well as the results of imaging and pathologic examinations and may help to raise awareness of this malformation for early recognition.

Relationships between body condition score, body weight and body measurements in alpacas.

BACKGROUND: The nutritional status in alpacas is often masked by their dense fibre coat. Its assessment is commonly approached by different body condition scores (BCS) that rely on manual palpation of defined anatomical regions. However, BCS is an important diagnostic tool to aid recognition of diseased South American camelids (SACs) and low BCS has been associated with conditions like anaemia and neutrophilia. For dose-dependent veterinary treatment, body weight (BW), that should be as accurate as possible, is required. As on-site weighing with scales is often not possible, BW can mostly only be roughly estimated. To date, it remains unclear whether BCS in alpacas provides reliable information on BW or the ratios of BW to body length commonly known as Body Mass Index (BMI) or Ponderal Index (PI). Equations to estimate BW based on body measurements are available in the literature. Nonetheless, respective equations were developed in growing alpacas or adult llamas and BCS was not included. RESULTS: To compare six different BCS approaches and to examine the relationship between BCS and BW, body measurements and BCS scores were recorded in a herd of 105 alpacas. The examined BCS approaches showed significant (p < 0.05) but poor to moderate positive correlations to BW, BMI or PI. A solely visual inspection of BCS, in contrast, was not correlated with BW, BMI or PI. Equations previously developed in other studies provided an accurate estimation of BW. Multiple linear regression showed that the accuracy in predicting BW could be further increased by adding BCS data and sex. CONCLUSION: Our observations indicate that most selected BCS approaches are not only important measures of nutritional status but can also be used to create more accurate models for BW calculation in alpacas. The study also supports the claim that a purely visual inspection of alpacas is not an adequate method to evaluate the nutritional status of these animals.

Rhododendron poisoning in alpacas (Vicugna pacos) in Northern Germany.

Poisoning is often suspected to be the origin of disease in South American camelids (SACs) by owners, but only in a few cases this assumption can be confirmed. In small ruminants, rhododendron poisoning is a common emergency for livestock veterinarians. However, this condition has rarely been reported in SACs so far. This paper provides information regarding clinical findings, hematology, clinical chemistry, and treatment of four alpacas after presumed intake of rhododendron leaves including pathological findings of one of the animals. Rhododendron leaves contain grayanatoxins that lead to hyperpolarization of excitable cells. Clinical signs that were observed in the presented alpacas comprised: salivation, dehydration, decreased motility of compartment 1, uncoordinated regurgitation, and cardiac arrhythmia. Clinical chemistry revealed that rhododendron poisoning was associated with metabolic acidosis and azotaemia, hyponatremia and hyperkalemia. Most striking macroscopic and histopathological findings included gastric ulceration, and renal infarcts along with inflammatory changes. Leaves of Rhododendron spp. were identified in the forestomach content of this animal. Affected animals were treated symptomatically as there is no specific antidote in rhododendron poisoning. This included parenteral rehydration, treatment of metabolic acidosis (infusion of sodium bicarbonate solution), and oral administration of activated charcoal to bind potential toxins. In addition, antibiotic treatment might be necessary to prevent aspiration pneumonia in case of uncoordinated regurgitation. Of the four animals, the worst affected alpaca was euthanized, one had minimal signs and two responded to supportive care and recovered. In conclusion, rhododendron poisoning might be fatal for alpacas in individual cases and therefore rhododendron bushes should not be placed in the habitat of SACs.

Detection of Coxiella burnetii in the mammary gland of a dairy goat.

The zoonotic bacterium Coxiella (C.) burnetii can be excreted by infected goats through birth products and milk. The detection of C. burnetii DNA in the mammary gland tissue of infected dairy goats and intermittent milk shedders has been reported, but confirmation of C. burnetii bacteria in the udder remained pending. The pathogen caused abortions in a 152-head dairy goat herd, resulting in the vaccination against C. burnetii of the entire herd with annual boosters. To monitor the C. burnetii shedding at herd level, monthly bulk tank milk (BTM) samples were analyzed using PCR (IS1111). Despite vaccination, C. burnetii DNA was detected in BTM samples within the first 16 months of the study. Therefore, individual milk samples were tested on four different occasions several months apart to identify potential intermittent milk shedders. Only one goat (#67455) tested positive three times. This goat was necropsied to investigate the presence of C. burnetii in the udder and other organs. PCR detected C. burnetii DNA solely in both mammary glands and the left teat cistern. Immunohistological examination identified C. burnetii antigen in mammary gland tissue, confirmed by the detection of C. burnetii bacteria in the mammary epithelial cells using fluorescence in situ hybridization. The removal of goat #67455 led to negative BTM samples until the end of the study. The findings demonstrate the occurrence of C. burnetii in the mammary gland of a naturally infected and vaccinated goat. The presence possibly contributed to intermittent milk shedding of goat #67455, and the mammary gland tissue may serve as a replicative niche for C. burnetii.

Hematologic parameters in female alpacas during age progression: a retrospective study.

Alpacas, like all camelids, have elliptical red blood cells (RBCs) in contrast to other mammals. This particular shape is important for increased osmotic resistance and stability. Age-related changes in the RBC count are known in other species, with alterations in both red and white blood cells being described. In alpacas, there are few data on age-related changes, and only a comparison of crias with adult animals. We characterized age-related hematologic changes in a study of 21 female alpacas from a research herd. A total of 87 records of clinically healthy alpacas of different ages were statistically analyzed retrospectively from the hematologic records over a nine-year period. Significant positive correlations of age with hemoglobin (Hb), HCT, MCV, MCH, neutrophils, platelet-to-lymphocyte ratio (PLR), and neutrophil-to-lymphocyte ratio (NLR) were found as well as significant negative correlations of age with lymphocytes in addition to lymphocyte-to-monocyte ratio (LMR). A paired comparison of eight older animals in the herd at three different ages also showed significant differences in the parameters Hb, HCT, MCV, MCH, MCHC, lymphocytes, eosinophils and neutrophils. Similar changes in hematologic parameters have been reported in other species and should be taken into account when interpreting hematologic results in alpacas.

Anaemia in South American camelids - an overview of clinical and laboratory diagnostics.

South American camelids (SACs) play an increasing role in veterinary care in Europe. Many alpacas or llamas presented to veterinarians suffer from anaemia, regularly with a packed cell volume (PCV) below 0.10 l/l, which is a life-threatening condition for the animals. This review article presents clinical and laboratory diagnostic tools for the diagnosis of anaemia in SACs. Clinical identification of anaemic animals can be performed by assessing the FAMACHA© score and the Body Condition Score (BCS), since anaemia in alpacas and llamas correlates with pale mucous membranes and a lowered BCS. Haematological examination of a blood sample can provide a more differentiated diagnosis of anaemia in SACs. A common finding is regenerative anaemia with an increased number of reticulocytes that is often caused by blood loss due to Haemonchus contortus. Changes in a blood smear from an alpaca or llama with regenerative anaemia may include normoblasts (nucleated red blood cells), anisocytosis, poikilocytosis, polychromasia, Howell-Jolly bodies or basophilic stippling. Furthermore, non-regenerative anaemia, often caused by trace element deficiency or cachexia, can also occur.

Body condition scoring in alpacas (Vicugna pacos) and llamas (Lama glama) - a scoping review.

Assessment of the body condition score (BCS) is an important tool to check the nutritional status of an animal, and thus can provide important information in herd management, but also in the veterinary examination of individual animals. In dairy farming, BCS has been recognized as an important parameter for a long time already, but over the years body condition scoring has also become more important for other species. Especially in South American camelids (SACs: alpacas and llamas), whose dense coat can hide a poor nutritional status for a long time, regular assessment of the BCS is recommended by different authors and organizations. To date, there are several instructions for the assessment of the BCS in SACs, which, however, differ significantly in some points. For a consistent approach, a total of 35 instructions for the BCS in SACs were compared in a scoping review. The spine, especially the lumbar vertebrae as well as the ribs were identified as the two main body sites recommended for assessing the BCS. Furthermore, the area between the front and rear leg and the pelvis were mentioned frequently; however, these body sites were discussed controversially. The paralumbar fossa, shoulder, and udder were only mentioned sporadically. Among the various scales, the 1-5 scale was mentioned most frequently, with the optimal BCS being 3. Nonetheless, physiological fluctuation due to breed, age, sex, pregnancy, and lactation have to be considered. The BCS should be assessed each time the animals are handled, but at least monthly. In contrast to body weight, the BCS reflects the nutritional status of an animal even after a single examination. The review showed that much of the available information on the assessment of BCS in SACs is based on practical experience rather than scientific evidence. Indeed, some of the assumptions made there might still have to be verified or modified.

Interdisciplinary studies on Coxiella burnetii: From molecular to cellular, to host, to one health research.

The Q-GAPS (Q fever GermAn interdisciplinary Program for reSearch) consortium was launched in 2017 as a German consortium of more than 20 scientists with exceptional expertise, competence, and substantial knowledge in the field of the Q fever pathogen Coxiella (C.) burnetii. C. burnetii exemplifies as a zoonotic pathogen the challenges of zoonotic disease control and prophylaxis in human, animal, and environmental settings in a One Health approach. An interdisciplinary approach to studying the pathogen is essential to address unresolved questions about the epidemiology, immunology, pathogenesis, surveillance, and control of C. burnetii. In more than five years, Q-GAPS has provided new insights into pathogenicity and interaction with host defense mechanisms. The consortium has also investigated vaccine efficacy and application in animal reservoirs and identified expanded phenotypic and genotypic characteristics of C. burnetii and their epidemiological significance. In addition, conceptual principles for controlling, surveilling, and preventing zoonotic Q fever infections were developed and prepared for specific target groups. All findings have been continuously integrated into a Web-based, interactive, freely accessible knowledge and information platform (www.q-gaps.de), which also contains Q fever guidelines to support public health institutions in controlling and preventing Q fever. In this review, we will summarize our results and show an example of how an interdisciplinary consortium provides knowledge and better tools to control a zoonotic pathogen at the national level.

[Diaphragmatic hernia in a female alpaca (Vicugna pacos)]. / Zwerchfellhernie bei einer Alpakastute (Vicugna pacos).

A 3-year-old pregnant female alpaca was noticed due to increased lying and decreased feed intake after shearing. Twelve days after shearing, she aborted an almost completely developed cria. After another two days, the alpaca was recumbent and subsequently died during transport to the clinic. A dissection of the carcass revealed a diaphragmatic hernia. Diaphragmatic hernias have been described in many different species, diagnosis can be made by radiography or ultrasound. Depending on the extent, surgical therapy may be successful.

Efficacy of flukicides against Fasciola hepatica and first report of triclabendazole resistance on German sheep farms.

Fasciola hepatica infections lead to severe health problems and production losses in sheep farming, if not treated effectively. Triclabendazole has been used extensively over decades due to its unique efficacy range against all definitive hostfluke stages but published data about the susceptibility of F. hepatica to anthelmintics in Germany are lacking. This study aimed to identify current F. hepatica infections in German sheep flocks by coproscopic examinations and to evaluate the efficacy of anthelmintics with a focus on triclabendazole in a field study conducted from 2020 to 2022. Initial screening included 71 sheep farms, many of them with known history of fasciolosis. In this highly biased sample set, the frequency of F. hepatica infection at individual sheep and farm level were 12.8% and 35.2%, respectively. Additionally, eggs of Paramphistominae were found at frequencies of 4.8% and 15.5% at individual sheep and farm level, respectively. Due to low egg shedding intensity, faecal egg count reduction (FECR) tests could only be conducted on a few farms. The efficacy of triclabendazole was tested on 11 farms and albendazole on one farm, including 3-53 sheep/farm. Individual faecal samples were collected before and two weeks after treatment to evaluate the FECR using the sedimentation or FLUKEFINDER® or a modified FLUKEFINDER® method. On all farms a coproantigen reduction test was conducted in parallel. Lacking efficacy of triclabendazole even at double dosage was shown on one farm associated with a high number of animal losses due to acute fasciolosis. On this farm, the Fasciola miracidium development test was additionally performed, revealing a high in vitro ovicidal activity of albendazole while closantel was effective in vivo. On all other farms, sufficient efficacy of triclabendazole was observed. In conclusion, triclabendazole resistance appears not to be widespread on German sheep farms but, when present, can have serious effects on animal health.

Teaching clinical hematology and leukocyte differentiation in veterinary medicine using virtual patients.

Due to contact restrictions imposed because of the COVID-19 pandemic, we created a novel digital course on the Moodle learning platform for winter term in 2020. In the clinical pathology course (CPC) with hematological content, third-year students were able to work independently on 10 extra digital cases of internal medicine involving eight different animal species as a compensation for the reduction in traditional microscopy exercises. Each case presented was initiated using an anamnesis, also the participants to generate a differential blood count based on digitized leukocytes, previously been photographed using a microscope camera. The cases were successive and increased in complexity, for example through the increase in the number of different cell types to be differentiated. The participants had the opportunity to evaluate the course through a final module to rate user-friendliness and acceptance. The total results of the participants in 2021 were analyzed descriptively, focusing on success rates, time spent on the tasks, and number of attempts. A total of 237 (= 96%) of 247 students completed all cases, each assessing 1033 photographed blood cells in sum. The mean processing time was 22.48 min for a differentiation and the students spent an average of 1.48 attempts on it. A voluntary feedback form was completed by 192 (= 78%) students, with more than 95% rating the course positively in 12 evaluation questions, and 29 of 33 comments (= 87.88%) providing positive statements in a comment box. Suggestions for improvement primarily included more explanations on erythrocyte morphologies, followed by adjusting the difficulty level and improving the presentational set-up. Slight improvements in results, time spent on processing the tasks, and the number of attempts indicated an achievement of routine and confidence during the course and were associated with an increase of competency. The positive feedback showed a high acceptance of the digital format and students evaluated the course as improving the quality of teaching when combined with practical exercises.

Two Years after Coxiella burnetii Detection: Pathogen Shedding and Phase-Specific Antibody Response in Three Dairy Goat Herds.

The infection dynamics of Coxiella (C.) burnetii were investigated in three dairy goat herds (A, B, and C) 2 years after the first pathogen detection. A total of 28 and 29 goats from herds A and B, and 35 goats from herd C, were examined. Sera were analyzed on three sampling dates using phase-specific serology. Pathogen shedding was assessed using post-partum vaginal swabs and monthly bulk tank milk (BTM) samples. Dust samples from a barn and milking parlor were also collected monthly. These samples were analyzed with PCR (target IS1111). In herd A, individual animals tested seropositive, while vaginal swabs, BTM, and most dust samples tested negative. Herds B and C exhibited high IgG phase I activity, indicating a past infection. In herd B, approximately two-thirds of the goats shed C. burnetii with vaginal mucus, and irregular positive results were obtained from BTM. Herd C had two positive goats based on vaginal swabs, and BTM tested positive once. Dust samples from herds B and C contained C. burnetii DNA, with higher quantities typically found in samples from the milking parlor. This study highlights the different infection dynamics in three unvaccinated dairy goat herds and the potential use of dust samples as a supportive tool to detect C. burnetii at the herd level.

Severe anaemia secondary to a perforated gastric ulcer in a male alpaca.

BACKGROUND: Anaemia is a common condition in alpacas and attributable to a variety of causes. Severe anaemia with a packed cell volume (PCV) less than 10% is frequently diagnosed, usually due to blood loss resulting from haemonchosis. Many South American camelids (SACs) also suffer from gastric ulcers, which are often associated with anaemia in other species. However, in alpacas and llamas, gastric ulcers usually do not lead to anaemia due to blood loss according to the current literature. There are no detailed clinical and laboratory data on this condition in the scientific literature so far. CASE PRESENTATION: We report on the case of a nine-year-old male alpaca that was presented to the clinic with suspected forestomach acidosis. The animal showed clinical signs of colic, hypothermia, tachypnea, tachycardia, pale mucous membranes, and died shortly after admission to the clinic. Laboratory diagnosis revealed a markedly decreased haematocrit (0.13 l/l), leucopaenia with band neutrophils, azotaemia, hypocalcaemia, hyperphosphataemia and vitamin D deficiency. Post-mortem examination revealed multiple ulcers in the first and third compartment with perforation of one ulcer in the first compartment, resulting in intraluminal blood loss and purulent peritonitis. CONCLUSIONS: To the authors' knowledge, this is the first detailed description of clinical and laboratory data of severe anaemia due to a perforated gastric ulcer in a SAC. Although the current literature suggests that severe blood loss due to gastric ulcers does not occur in SACs, this condition should be considered as a possible differential diagnosis in anaemic animals. Clinical indicators can be colic and pale mucous membranes.

Humoral and cellular immune responses in sheep following administration of different doses of an inactivated phase I vaccine against Coxiella burnetii.

An inactivated Coxiella burnetii Phase I (PhI) vaccine (Coxevac®) is licensed in several European countries for goats and cattle to prevent coxiellosis. The vaccine is also applied to sheep, although detailed information about the ovine immune response and vaccine dose is missing. Eighteen gimmers from a C. burnetii unsuspected flock were randomly divided into three groups of six. Group 1 (Cox1) and 2 (Cox2) were vaccinated twice with 1 ml and 2 ml Coxevac®, respectively, three weeks apart (primary vaccination). The same procedure was applied with Cox3 (2 ml sodium chloride, control group). A third injection (booster) was performed after nine months. Potential side effects were determined by measuring the rectal body temperature and skin thickness at the injection site. Blood samples were collected to detect phase-specific IgM and IgG antibodies and interferon-É£ (IFN-É£) release by immunofluorescence assay and ELISAs, respectively. Moreover, a cell infection neutralization assay determined the appearance of neutralizing sera. Body temperatures increased for one day post vaccination, and the skin swelled only slightly. Regardless of the vaccine volume, immunized sheep reacted first with an IgM and IgG PhII response. Ten weeks after the primary vaccination, IgG PhI antibodies predominated. Boosting eight months after primary vaccination resulted in a robust IgG PhI increase and strong IFN-É£ response. In the vaccinated animals, the neutralizing effect is more widespread after the administration of 1 ml than after the treatment with 2 ml. In summary, differences between 1 and 2 ml Coxevac® are minor, and a vaccine volume of 1 ml seems to be sufficient. A booster after the primary vaccination is apparently necessary to stimulate the cell-mediated immune response in naïve sheep.

Seroprevalence of tick-borne encephalitis virus in wild and domestic animals in northern Germany.

Tick-borne encephalitis virus (TBEV) is a tick-transmitted flavivirus, which can infect humans and animals, sometimes even with a fatal outcome. Since many decades, TBEV is endemic in southern Germany, while only sporadic occurrence has been noted in northern parts of the country so far. Nevertheless, autochthonous human clinical cases are increasing in the federal state of Lower Saxony in north-western Germany, and several natural foci of TBEV transmission have recently been detected in this federal state. In order to shed more light on the current distribution of TBEV in Lower Saxony, the present study examined blood samples from wild and domestic animals for antibodies against TBEV. Overall, samples from 4,085 animals were tested by ELISA, including wild boar (N = 1,208), roe deer (N = 149), red deer (N = 61), fallow deer (N = 18), red foxes (N = 9), nutria (N = 9), raccoon dogs (N = 3), raccoons (N = 3), badgers (N = 1), European pine martens (N = 1), horses (N = 574), sheep (N = 266), goats (N = 67), dogs (N = 1,317) and cats (N = 399). Samples with an ELISA result of ≥60 Vienna units (VIEU)/ml were subjected to confirmatory serum neutralization tests (SNT). In total, 343 of 4,085 (8.4%) animals tested positive for anti-TBEV-IgG by ELISA, of which 60 samples were confirmed by SNT. Samples of 89 animals showed a cytotoxic effect in the SNT and were excluded from seroprevalence calculation, resulting in an overall seroprevalence of 1.5% (60/3,996). Seroprevalence was higher among wild animals (wild boar: 2.9% [34/1,190], roe deer: 2.7% [4/149], red deer: 1.7% [1/60], fallow deer: 5.6% [1/18]) than among domestic animals (dogs: 1.1% [15/1,317], horses: 0.8% [4/505], sheep: 0.4% [1/266]). No anti-TBEV-antibodies were detected in the other wild animal species as well as goats and cats. A notable clustering of positive samples was observed in districts where TBEV transmission foci have been described. Further clusters in other districts suggest the existence of so far undetected transmission foci, underlining the fact that both wild and domestic animals are useful sentinels for monitoring the spread of TBEV.

Detection and Characterization of Alongshan Virus in Ticks and Tick Saliva from Lower Saxony, Germany with Serological Evidence for Viral Transmission to Game and Domestic Animals.

The newly discovered group of Jingmenviruses has been shown to infect a wide range of hosts and has been associated with febrile illness in humans. During a survey for Jingmenviruses in ticks from Lower Saxony, Germany, Alongshan virus (ALSV) was identified in Ixodes spp. ticks. Additional virus screenings revealed the presence of ALSV in the bodies and saliva of ticks collected at several locations in Lower Saxony. Vector competence studies that included Ixodes ricinus and Dermacentor reticulatus validated the replication of ALSV within those tick species. In vitro feeding experiments with ALSV-injected Ixodes ricinus demonstrated effective viral transmission during blood feeding. To evaluate the potential viral transmission during a natural blood meal, sera from wild game and domestic animals were investigated. One serum sample from a red deer was found to be positive for ALSV RNA, while serological screenings in game and domestic animals revealed the presence of ALSV-specific antibodies at different locations in Lower Saxony. Overall, those results demonstrate the broad distribution of ALSV in ticks in Lower Saxony and hypothesize frequent exposure to animals based on serological investigations. Hence, its potential risk to human and animal health requires further investigation.

Co-exposure to Anaplasma spp., Coxiella burnetii and tick-borne encephalitis virus in sheep in southern Germany.

The intracellular bacteria Anaplasma spp. and Coxiella burnetii and the tick-borne encephalitis virus (TBEV) are tick-transmitted pathogens circulating in the southern German sheep population. Knowledge of interaction among Anaplasma spp., C. burnetii and TBEV in sheep is lacking, but together they might promote and reinforce disease progression. The current study aimed to identify co-exposure of sheep to Anaplasma spp., C. burnetii and TBEV. For this purpose, 1,406 serum samples from 36 sheep flocks located in both southern German federal states, Baden-Wuerttemberg and Bavaria, were analysed by ELISAs to determine the antibody levels of the three pathogens. Inconclusive and positive results from the TBEV ELISA were additionally confirmed by a serum neutralisation assay. The proportion of sheep with antibodies against Anaplasma spp. (47.2%), C. burnetii (3.7%) and TBEV (4.7%) differed significantly. Significantly more flocks with Anaplasma spp. seropositive sheep (91.7%) were detected than flocks with antibodies against TBEV (58.3%) and C. burnetii (41.7%), but there was no significant difference between the number of flocks which contained TBEV and C. burnetii seropositive sheep. Seropositivity against at least two pathogens was detected in 4.7% of sheep from 20 flocks. Most co-exposed sheep had antibodies against Anaplasma spp./TBEV (n = 36), followed by Anaplasma spp./C. burnetii (n = 27) and Anaplasma spp./C. burnetii/TBEV (n = 2). Only one sheep showed an immune response against C. burnetii and TBEV. Flocks with sheep being positive against more than one pathogen were widely distributed throughout southern Germany. The descriptive analysis revealed no association between the antibody response of the three pathogens at animal level. Taking the flocks as a cluster variable into account, the exposure to TBEV reduced the probability of identifying C. burnetii antibodies in sheep significantly (odds ratio 0.46; 95% confidence interval 0.24-0.85), but the reason for this is unknown. The presence of Anaplasma spp. antibodies did not influence the detection of antibodies against C. burnetii and TBEV. Studies under controlled conditions are necessary to evaluate any possible adverse impact of co-exposure to tick-borne pathogens on sheep health. This can help to clarify rare disease patterns. Research in this field may also support the One Health approach due to the zoonotic potential of Anaplasma spp., C. burnetii and TBEV.

The influence of different examiners on the Body Condition Score (BCS) in South American camelids-Experiences from a mixed llama and alpaca herd.

Particularly in unshorn llamas and alpacas with a dense fiber coat, changes in body condition often remain undetected for a long time. Manual palpation of the lumbar vertebrae is hence a simple and practical method for the objective assessment of body condition in South American camelids (SAC). Depending on tissue coverage, a body condition score (BCS) of 1 (emaciated) to 5 (obese) with an optimum of 3 is assigned. To date, there is a lack of detailed information on the comparability of the results when the BCS in llamas or alpacas is assessed by different examiners. Reliability of BCS assessment of 20 llamas and nine alpacas during a veterinary herd visit by six examiners was hence evaluated in this study. A gold standard BCS (gsBCS) was calculated from the results of the two most experienced examiners. The other examiners deviated by a maximum of 0.5 score points from the gsBCS in more than 80% of the animals. Inter-rater reliability statistics between the assessors were comparable to those in body condition scoring in sheep and cattle (r = 0.52-0.89; τ = 0.43-0.80; κw = 0.50-0.79). Agreements were higher among the more experienced assessors. Based on the results, the assessment of BCS in SAC by palpation of the lumbar vertebrae can be considered as a simple and reproducible method to reliably determine nutritional status in llamas and alpacas.