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A new betacoronavirus—Middle East respiratory syndrome coronavirus (MERS-CoV)—has been identified in patients with severe acute respiratory infection. Although related viruses infect bats, molecular clock analyses have been unable to identify direct ancestors of MERS-CoV. Anecdotal exposure histories suggest that patients had been in contact with dromedary camels or goats. We investigated possible animal reservoirs of MERS-CoV by assessing specific serum antibodies in livestock. We took sera from animals in the Middle East (Oman) and from elsewhere (Spain, Netherlands, Chile). Cattle (n=80), sheep (n=40), goats (n=40), dromedary camels (n=155), and various other camelid species (n=34) were tested for specific serum IgG by protein microarray using the receptor-binding S1 subunits of spike proteins of MERS-CoV, severe acute respiratory syndrome coronavirus, and human coronavirus OC43. Results were confirmed by virus neutralisation tests for MERS-CoV and bovine coronavirus. 50 of 50 (100%) sera from Omani camels and 15 of 105 (14%) from Spanish camels had protein-specific antibodies against MERS-CoV spike. Sera from European sheep, goats, cattle, and other camelids had no such antibodies. MERS-CoV neutralising antibody titres varied between 1/320 and 1/2560 for the Omani camel sera and between 1/20 and 1/320 for the Spanish camel sera. There was no evidence for cross-neutralisation by bovine coronavirus antibodies. MERS-CoV or a related virus has infected camel populations. Both titres and seroprevalences in sera from different locations in Oman suggest widespread infection. European Union, European Centre For Disease Prevention and Control, Deutsche Forschungsgemeinschaft.

This study assesses the use of fructosamine as a diagnostic tool for hyperglycemia in alpacas in view of their sensitivity to stress and susceptibility to conditions like lipid mobilization syndrome. Plasma fructosamine, like in diagnosing diabetes in cats and dogs, can reveal long-term blood glucose trends, differentiating stress-induced spikes from persistent diabetic hyperglycemia. In 125 alpacas presented as patients of a veterinary clinic, plasma glucose and fructosamine concentrations were compared for correlations with findings of the general clinical examination, laboratory parameters, demographic data, and a behavioral stress assessment processed by using principal component analysis. Hyperglycemia was observed on admission of 71% (89/125) of the animals. This was significantly associated with a higher concentration of serum cortisol and a higher behavioral stress scoring. Fructosamine above the reference limit was detected in only 15% (13/89) of the hyperglycemic individuals. In addition to a positive correlation of fructosamine to glucose concentration, positive relationships with different plasma proteins were detected. A relationship to stress parameters was not observed. These findings underscore stress as a significant trigger for hyperglycemia in alpacas and suggest fructosamine as a valuable parameter for distinguishing between stress-induced and diabetic hyperglycemia. However, the dependence of fructosamine formation on total plasma protein concentration should be considered to avoid misinterpretation.

White blood cell (WBC) ratios are used as diagnostic markers for various inflammatory or tumor diseases as well as stress in a broad range of species. The aim of this work was to provide data on five WBC ratios (neutrophil-to-lymphocyte ratio [NLR], band neutrophil-to-lymphocyte ratio [BLR], band neutrophil-to-neutrophil-to-lymphocyte ratio [BNLR], band neutrophil-to-neutrophil ratio [BNR] and lymphocyte-to-monocyte ratio [LMR]) in South American camelids (SAC) and characterize their association with demographic and important diagnostic parameters. Medical records of 307 SAC (275 alpacas, 32 llamas) that were presented at a veterinary teaching hospital were evaluated retrospectively. WBC ratios were calculated based on hematologic results of the initial blood samples. The influence of species, sex, age, body condition score, WBC count, and anemia on those ratios was investigated using descriptive statistics and generalized linear models. NLR, BLR and LMR were found to be significantly influenced by age and WBC count. Associations of individual WBC ratios with species, nutritional status or an anemic condition could be detected. NLR was 4.32; 2.31–7.81 (median; IQR), BLR 0.24; 0.07–0.87, BNLR 3.66 × 10 –3 ; 1.17 × 10 –3 − 14.20 × 10 –3 , BNR 0.06; 0.02–0.15 and LMR was 7; 3.54–14.67. Our data might serve as a basis for further studies on WBC ratios in SAC. The animals in this study showed a variety of underlying diseases. It should hence be noted that these values are orientation values and provide a representative overview of conditions in a clinic, but are not suitable as reference values for healthy animals.

The adequate transfer of passive immunity is a critical factor in neonatal development and survivability. Although well documented in the dairy and equine industries, the recognition of inadequate immunoglobulin transfer on-farm and its impact on the ability of alpaca cria to thrive is largely unknown. Colostrum samples were collected from female alpaca within 24 h of parturition by the owners and whole blood collected from cria by the investigators between 1 and 7 days of age. Direct IgG concentration of milk and serum was determined using radial immunodiffusion assay (RID) and was indirectly estimated using optical and digital Brix refractometry for total solids and clinical refractometry for total serum protein. There was a strong correlation between optical and digital Brix refractometry, and colostral IgG concentration determined by RID. There was a moderate correlation between serum IgG concentration determined by RID and total serum protein in crias. Optical and digital Brix refractometry for colostral IgG estimation and total serum protein for serum IgG estimation are reliable, accurate and easy-to-use tools that can be used on-farm by trained, competent technicians to assess a failure of passive transfer in alpacas. A pilot study at one property only was performed, due to COVID-19 travel restriction interference. Further research is required to determine the reference intervals for these tools to be practical.

Background Mycoplasma haemolamae (Mhl) and gastrointestinal nematodes can cause anemia in camelids. Control programs aim to suppress parasitism without promoting anthelminthic resistance, but few evidence‐based guidelines define acceptable parasite loads in camelids. Hypothesis/Objectives In clinically healthy nonanemic camelids, compare erythrocyte variables to Mhl real‐time PCR status and to fecal egg count (FEC). Determine the FEC threshold above which erythrocyte variables are consistently below reference interval medians. Animals One hundred fourteen client‐owned adult alpacas and llamas. Methods In a cross‐sectional study, whole blood in ethylenediaminetetraacetic acid (EDTA) was assessed for packed cell volume (PCV) by centrifugation, erythrocyte count (RBC), and hemoglobin concentration (HGB) using an ADVIA120 analyzer, and Mhl using real‐time PCR. Trichostrongyle eggs per gram (epg) were counted by modified McMaster test on freshly collected feces. Significant differences in erythrocyte variables based on Mhl status and FEC thresholds were assessed by independent t test and one‐way ANOVA, respectively. Results Packed cell volume, RBC, and HGB were not significantly different between Mhl‐positive and Mhl‐negative animals, but were significantly lower in animals with FEC >1000 epg compared to those with <500 epg. All animals with FEC >600 epg had RBC and HGB below the reference interval median. All animals with FEC >750 epg had PCV below the reference interval median. Conclusions and Clinical Importance In healthy nonanemic camelids, positive Mhl PCR is not associated with lower erythrocyte variables and such animals may not warrant treatment. Fecal egg count >600‐750 epg has a negative effect on erythrocyte variables, and may be a guide for deworming protocols.

ABSTRACT Objective Anti‐Müllerian hormone (AMH) has become an important hormonal parameter for the detection of gonadal tissue and for the diagnosis of gonadal functions and pathologies. To our knowledge, there is currently no homologous test for AMH measurements in South American camelids (SACs). Therefore, the objective of the present study was to determine serum AMH concentrations in postpubertal male alpacas and, for the first time, in llamas, using the Elecsys AMH assay kit that has not previously been tested in these species. To obtain indications of the specificity of this method in SAC, measurements were carried out in male gelding in which concentrations below the detection limit were to be expected. Methods In this context, 37 blood samples collected by jugular venipuncture from 21 alpacas and 16 llamas were used. The obtained blood was centrifuged at 3000 g for 20 min, and the serum was stored in Eppendorf tubes at −20°C until AMH concentrations were measurement. The measurement of AMH levels was conducted in a commercial diagnostic laboratory (Laboklin, Bad Kissingen, Germany) using the electrochemiluminescence immunoassay kit Elecsys AMH run on the fully automated Cobas e 601 analyser (Roche Diagnostics Deutschland GmbH, Mannheim). The AMH test had a minimum detection limit of 0.01 ng/mL and a maximum detection limit of 23 ng/mL. The intra‐assay coefficient of variation is between 2.7% and 3.3%. Results Blood serum AMH levels ranged between 4.10 and 22 ng/mL (median: 9.80 ng/mL) and 1.79 and 10.05 ng/mL (median: 4.00) in intact alpacas (age: 6.30 ± 2.71 years; n = 10) and llamas (age: 5.50 ± 4.34; n = 8), respectively, and were significantly different between samples obtained from the two species (p < 0.05). Correlation analyses regarding an age dependence of AMH concentrations yielded negative correlation coefficients for both species but non‐significant p values (alpaca: r = −0.165, p = 0.649; llama: r = −0.547, p = 0.160). In alpaca (n = 11) and llama geldings (n = 8), blood serum AMH levels were below 0.01 ng/mL (p < 0.001). These results prove that the antibodies used in the Elecsys AMH assay significantly and specifically cross‐react with SAC AMH. Conclusions In gelding llamas and alpacas, AMH concentrations were below the limit of detection (<0.01 ng/mL), which was significantly lower compared to intact animals (p < 0.001). The Elecsys AMH assay is therefore considered a suitable method for detecting gonadal tissue in SAC. Serum AMH values were different and statistically significant between the two species (p < 0.05).

The objective of this study was to develop and compare the performance of laboratory grade and portable attenuated total reflectance infrared (ATR-IR) spectroscopic approaches in combination with partial least squares regression (PLSR) for the rapid quantification of alpaca serum IgG concentration, and the identification of low IgG (<1000 mg/dL), which is consistent with the diagnosis of failure of transfer of passive immunity (FTPI) in neonates. Serum samples (n = 175) collected from privately owned, healthy alpacas were tested by the reference method of radial immunodiffusion (RID) assay, and laboratory grade and portable ATR-IR spectrometers. Various pre-processing strategies were applied to the ATR-IR spectra that were linked to corresponding RID-IgG concentrations, and then randomly split into two sets: calibration (training) and test sets. PLSR was applied to the calibration set and calibration models were developed, and the test set was used to assess the accuracy of the analytical method. For the test set, the Pearson correlation coefficients between the IgG measured by RID and predicted by both laboratory grade and portable ATR-IR spectrometers was 0.91. The average differences between reference serum IgG concentrations and the two IR-based methods were 120.5 mg/dL and 71 mg/dL for the laboratory and portable ATR-IR-based assays, respectively. Adopting an IgG concentration <1000 mg/dL as the cut-point for FTPI cases, the sensitivity, specificity, and accuracy for identifying serum samples below this cut point by laboratory ATR-IR assay were 86, 100 and 98%, respectively (within the entire data set). Corresponding values for the portable ATR-IR assay were 95, 99 and 99%, respectively. These results suggest that the two different ATR-IR assays performed similarly for rapid qualitative evaluation of alpaca serum IgG and for diagnosis of IgG <1000 mg/dL, the portable ATR-IR spectrometer performed slightly better, and provides more flexibility for potential application in the field.

Background The purpose of the study was to determine if the effect of llama OIF on LH secretion is mediated by stimulation of the hypothalamus or pituitary gland. Methods Using a 2-by-2 factorial design to examine the effects of OIF vs GnRH with or without a GnRH antagonist, llamas with a growing ovarian follicle greater than or equal to 8 mm were assigned randomly to four groups (n = 7 per group) and a) pre-treated with 1.5 mg of GnRH antagonist (cetrorelix acetate) followed by 1 mg of purified llama OIF, b) pre-treated with 1.5 mg of cetrorelix followed by 50 micrograms of GnRH, c) pre-treated with a placebo (2 ml of saline) followed by 1 mg of purified llama OIF or d) pre-treated with a placebo (2 ml of saline) followed by 50 micrograms of GnRH. Pre-treatment with cetrorelix or saline was given as a single slow intravenous dose 2 hours before intramuscular administration of either GnRH or OIF. Blood samples for LH measurement were taken every 15 minutes from 1.5 hours before to 8 hours after treatment. The ovaries were examined by ultrasonography to detect ovulation and CL formation. Blood samples for progesterone measurement were taken every-other-day from Day 0 (day of treatment) to Day 16. Results Ovulation rate was not different (P = 0.89) between placebo+GnRH (86%) and placebo+OIF groups (100%); however, no ovulations were detected in llamas pre-treated with cetrorelix. Plasma LH concentrations surged (P < 0.01) after treatment in both placebo+OIF and placebo+GnRH groups, but not in the cetrorelix groups. Maximum plasma LH concentrations and CL diameter profiles did not differ between the placebo-treated groups, but plasma progesterone concentrations were higher (P < 0.05), on days 6, 8 and 12 after treatment, in the OIF- vs GnRH-treated group. Conclusion Cetrorelix (GnRH antagonist) inhibited the preovulatory LH surge induced by OIF in llamas suggesting that LH secretion is modulated by a direct or indirect effect of OIF on GnRH neurons in the hypothalamus.

Background South American camelids in the United States have rapidly developed into an important agricultural industry in need of veterinary services. Pain management is challenging in camelids because there are no drugs currently approved by the U.S. Food and Drug Administration for use in these species. Dosage regimens used for many therapeutic drugs have been extrapolated from other ruminants; however, the pharmacokinetics, in camelids, may differ from those of other species. Studies investigating the pharmacokinetics of cyclooxygenase-2 (COX-2) selective non-steroidal anti-inflammatory drugs in camelids are deficient in the published literature. Six adult llamas (121- 168 kg) were administered either a 1 mg/kg dose of oral or a 0.5 mg/kg dose of IV meloxicam in a randomized cross-over design with an 11 day washout period between treatments. Plasma samples collected up to 96 hours post-administration were analyzed by high pressure liquid chromatography and mass spectrometry detection (HPLC-MS) followed by non-compartmental pharmacokinetic analysis. Results A mean peak plasma concentration (C MAX ) of 1.314 μg/mL (Range: 0.826 – 1.776 μg/mL) was recorded at 21.4 hours (Range: 12.0 – 24.0 hours) with a half-life (T ½ λ z ) of 22.7 hours (Range: 18.0 – 30.8 hours) after oral meloxicam administration. In comparison, a half-life (T ½ λ z ) of 17.4 hours (Range: 16.2 – 20.7 hours) was demonstrated with IV meloxicam administration. The oral bioavailability (F) of meloxicam (dose normalized) was 76% (Range: 48 – 92%). No adverse effects associated with either treatment modality were observed in the llamas. Conclusions The mean bioavailability (F) of oral meloxicam was 76% indicating a high degree of gastrointestinal absorption. Plasma meloxicam concentrations >0.2 μg/mL were maintained for up to 72 h after oral administration; >0.2 μg/mL is considered to be the concentration of meloxicam required for analgesic effects in other species such as the horse. These data suggest that a single dosage of oral meloxicam at 1 mg/kg could potentially maintain therapeutic concentrations in plasma for up to 3 days in adult llamas.

The aim of this study was to establish reference interval for biochemical parameters in blood of alpacas on the basis of large population of clinically healthy animals, and to determine the influence of sex, age and season on nitrogen and lipid metabolites, enzymes, electrolytes, vitamins and minerals in blood of alpacas. Blood samples were collected from 311 alpacas (61 males and 201 females >6 months of age and 49 crias (21 males and 28 females) ⩽6 months of age). Selected farms were located in Central Europe (Czech Republic and Germany). We determined 24 biochemical parameters from blood serum. We performed the comparison of results by the sex of animals and for the older group also the comparison of the results with regard to the season, respectively, to the feeding period. We found no highly significant difference (P<0.01) between males and females with the exception of γ-glutamyl transferase (GGT), alkaline phosphatase (ALP) and cholesterol. We found 15 significantly different parameters between the group of crias 6 months of age and the older alpacas. Based on our findings we suggest for most parameters to use different reference intervals (especially ALP, cholesterol, total protein, globulin, non-esterified fatty acids (NEFA), GGT and phosphorus) for the two above-mentioned age groups. Another important finding is the differences between some parameters in older group of alpacas in summer/winter feeding period. Animals in the summer feeding period have higher values of parameters related to fat mobilization (β-hydroxybutyrate, NEFA) and liver metabolism (bilirubin, alanine aminotransferase). The winter period with increased feeding of supplements with higher amount of fat, vitamins and minerals is characteristic by increased values of cholesterol, triglycerides, vitamins A and E, and some minerals (K, Ca, Mg and Cl) in blood serum. Clinical laboratory diagnosis of metabolic disturbances may be improved with use of age-based reference values and with consideration of seasonal differences.