Direct quantitative real‐time PCR assay for detection of the emerging pathogen Neonectria neomacrospora

The epidemic outbreak in northern Europe of Neonectria neomacrospora, the causal agent of dieback in Abies spp., led the European and Mediterranean Plant Protection Organization (EPPO) to include the pathogen on its alert list in 2017. Effective monitoring of this pathogen calls for a rapid and sensitive method of identification and quantification. A probe‐based real‐time PCR (qPCR) assay based on the β‐tubulin gene was developed for the detection and quantification of N. neomacrospora in infected wood samples, and directly for ascospores. This study presents the first published species–specific molecular detection assay for N. neomacrospora. The analytical specificity was validated on taxonomically closely related fungal species as well as on 18 fungal species associated with the host (Abies sp.). The analytical sensitivity was tested on naturally infected wood, on purified pathogen DNA in a matrix of host DNA and on N. neomacrospora ascospores for detection of airborne inoculum. The latter was tested both with a DNA extraction step prior to qPCR and without DNA extraction by direct qPCR on collected ascospores. The assay was specific to N. neomacrospora, with a sensitivity of 130 fg purified DNA, or 10 ascospores by direct qPCR. Omitting DNA extraction and amplifying directly on unpurified ascospores improved assay sensitivity significantly.     

Effects of Salinity,Commercial Salts,and Water Type on Cultivation of the Cryptophyte Microalgae Rhodomonas salina and the Calanoid Copepod Acartia tonsa

Marine aquaculture facilities positioned far from the sea need access to seawater (SW); hence, commercial salts are often the chosen solution. In marine hatcheries, most fish larvae require live feed (zooplankton) that are in turn fed with microalgae. The objective of this research was to investigate the applicability of commercial salts and clarify the potential effects on the cultivation of the microalga Rhodomonas salina and the copepod Acartia tonsa. Three commercial salts were tested, Red Sea Salt (RS), Red Sea – Coral Pro Salt (CP), and Blue Treasure Salt. R. salina was cultured at salinities of 10, 20, and 30 psu resulting in equal growth rates at salinities 20 and 30 in SW and RS mixed with deionized (DI) water. The optimum salinity for R. salina was 29 psu. For A. tonsa eggs, we observed highest hatching success in 30 psu with CP or RS mixed with DI water. The egg hatching success was not affected by salinities 15–40 and optimal hatching was obtained at 27 psu. Results confirm it was possible to use commercial salts for rearing of both R. salina and A. tonsa, widening the application of these species for aquaculture facilities without access to SW.     

Ante mortem diagnosis of paratuberculosis: a review of accuracies of ELISA, interferon-gamma assay and faecal culture techniques

Infections with Mycobacterium avium subsp. paratuberculosis (MAP) can be latent for years without affecting the animal, but the animal may become infectious or clinical at some point. Diagnosis of paratuberculosis can be a challenge primarily in latent stages of the infection, and different diagnosis interpretations are usually required by the variety of decision makers. The objective of this paper was to provide a critical review of reported accuracies of ELISA tests, interferon-gamma assays (IFN-gamma) and faecal culture (FC) techniques used for diagnosis of three defined target conditions: MAP infected, MAP infectious and MAP affected animals. For each animal species, target condition and diagnostic test-type, sensitivities (Se) and specificities (Sp) were summarised based on a systematic, critical review of information in literature databases. The diagnostic test information often varied substantially for tests of the same type and make, particularly ELISA, which was the most frequently reported test-type. Comparison of the various tests accuracies was generally not possible, but stratification of test-evaluations by target condition improved the interpretation of the test accuracies. Infectious and affected animals can often be detected, but Se for infected animals is generally low. A main conclusion of the review was that the quality of design, implementation and reporting of evaluations of tests for paratuberculosis is generally poor. Particularly, there is a need for better correspondence between the study population and target population, i.e. the subjects chosen for test evaluation should reflect the distribution of animals in the population where the test is intended to be used.     

Low ratios of sodium to potassium in the serum of 238 dogs

Serum sodium:potassium (Na:K) ratios are often reported in biochemical studies of dogs, although their value has not been assessed. The aims of this study were to identify diseases associated with a low Na:K ratio in dogs and to compare their prevalence with the prevalence in dogs from the same referral hospital with normal Na:K ratios. A total of 238 dogs with a Na:K ratio less than 27 were identified from medical records. Sample contamination with edta was suspected in 74 cases (31 per cent) and these and two cases that had been supplemented with potassium were removed from the analysis. The remaining 162 cases and 147 control dogs were divided into five categories depending on the organ system affected. Among the cases there were significantly more in the endocrine category than among the control dogs. Hypoadrenocorticism was the most single common cause of a low Na:K ratio and affected 27 (16.7 per cent) of the cases. Other clinical problems associated with low Na:K ratios included different urogenital, cardiorespiratory and gastrointestinal diseases.     

Impacts of prenatal nutrition on animal production and performance: a focus on growth and metabolic and endocrine function in sheep

The concept of foetal programming(FP) originated from human epidemiological studies, where foetal life nutrition was linked to health and disease status later in life. Since the proposal of this phenomenon, it has been evaluated in various animal models to gain further insights into the mechanisms underlying the foetal origins of health and disease in humans. In FP research, the sheep has been quite extensively used as a model for humans. In this paper we will review findings mainly from our Copenhagen sheep model, on the implications of late gestation malnutrition for growth, development, and metabolic and endocrine functions later in life, and discuss how these implications may depend on the diet fed to the animal in early postnatal life. Our results have indicated that negative implications of foetal malnutrition, both as a result of overnutrition and, particularly, late gestation undernutrition, can impair a wide range of endocrine functions regulating growth and presumably also reproductive traits. These implications are not readily observable early in postnatal life, but are increasingly manifested as the animal approaches adulthood. No intervention or cure is known that can reverse this programming in postnatal life. Our findings suggest that close to normal growth and slaughter results can be obtained at least until puberty in animals which have undergone adverse programming in foetal life, but manifestation of programming effects becomes increasingly evident in adult animals.Due to the risk of transfer of the adverse programming effects to future generations, it is therefore recommended that animals that are suspected to have undergone adverse FP are not used for reproduction. Unfortunately, no reliable biomarkers have as yet been identified that allow accurate identification of adversely programmed offspring at birth,except for very low or high birth weights, and, in pigs, characteristic changes in head shape(dolphin head). Future efforts should be therefore dedicated to identify reliable biomarkers and evaluate their effectiveness for alleviation/reversal of the adverse programming in postnatal life. Our sheep studies have shown that the adverse impacts of an extreme, high-fat diet in early postnatal life, but not prenatal undernutrition, can be largely reversed by dietary correction later in life. Thus, birth(at term) appears to be a critical set point for permanent programming in animals born precocial,such as sheep. Appropriate attention to the nutrition of the late pregnant dam should therefore be a priority in animal production systems.     

Explanation and Elaboration Document for the STROBE‐Vet Statement: Strengthening the Reporting of Observational Studies in Epidemiology – Veterinary Extension

The STROBE (Strengthening the Reporting of Observational Studies in Epidemiology) statement was first published in 2007 and again in 2014. The purpose of the original STROBE was to provide guidance for authors, reviewers and editors to improve the comprehensiveness of reporting; however, STROBE has a unique focus on observational studies. Although much of the guidance provided by the original STROBE document is directly applicable, it was deemed useful to map those statements to veterinary concepts, provide veterinary examples and highlight unique aspects of reporting in veterinary observational studies. Here, we present the examples and explanations for the checklist items included in the STROBE‐Vet Statement. Thus, this is a companion document to the STROBE‐Vet Statement Methods and process document, which describes the checklist and how it was developed.     

Aortic rupture causing cardiac tamponade in a 24‐day‐old Friesian colt with concurrent colonic Chlamydiosis and Balantidiasis

A 24‐day‐old Friesian colt died suddenly and a physical examination the morning the foal died showed no abnormalities and serum IgG levels >8.0 g/l. Necropsy examination revealed haemopericardium and a 2 cm transverse tear at the root of the aorta. The foal was also found to have Chlamydophila spp. in the epithelium and Balantidium coli on the mucosal surface of the large colon. An aortic rupture is a novel finding in a foal, colonic Chlamydiosis has not been previously reported in horses and Balantidium coli has not been reported in equids in North America.     

Platelet function in dogs: breed differences and effect of acetylsalicylic acid administration

BACKGROUND: Clinical studies investigating platelet function in dogs have had conflicting results that may be caused by normal physiologic variation in platelet response to agonists. OBJECTIVES: The objective of this study was to investigate platelet function in clinically healthy dogs of 4 different breeds by whole-blood aggregometry and with a point-of-care platelet function analyzer (PFA-100), and to evaluate the effect of acetylsalicylic acid (ASA) administration on the results from both methods. METHODS: Forty-five clinically healthy dogs (12 Cavalier King Charles Spaniels [CKCS], 12 Cairn Terriers, 10 Boxers, and 11 Labrador Retrievers) were included in the study. Platelet function was assessed by whole-blood aggregation with ADP (1, 5, 10, and 20 microM) as agonist and by PFA-100 using collagen and epinephrine (Col + Epi) and Col + ADP as agonists. Plasma thromboxane B(2) concentration was determined by an enzyme immunoassay. To investigate the effect of ASA, 10 dogs were dosed daily (75 or 250 mg ASA orally) for 4 consecutive days. RESULTS: A higher platelet aggregation response was found in CKCS compared to the other breeds. Longer PFA-100 closure time (Col + Epi) was found in Cairn Terriers compared to Boxers. Plasma thromboxane B(2) concentration was not statistically different between groups. Administration of ASA prolonged the PFA-100 closure times, using Col + Epi (but not Col + ADP) as agonists. Furthermore, ASA resulted in a decrease in whole-blood platelet aggregation. CONCLUSIONS: Platelet function is influenced by breed, depending upon the methodology applied. However, the importance of these breed differences remains to be investigated. The PFA-100 method with Col + Epi as agonists, and ADP-induced platelet aggregation appear to be sensitive to ASA in dogs.     

The degree of starch phosphorylation as influenced by phosphate deprivation of potato (Solanum tuberosum L.) plants

Summary The levels of starch phosphorylation in potato tubers were investigated in relation to fertilization with phosphorus (P) of field-grown and greenhouse-grown plants. The field-grown plants received 0, 15 or 30 kg P ha⁻¹. Starch from plants grown without P-fertilizer contained 15.6 nmol P (mg starch)⁻¹ whereas starch from plants grown with 30 kg P ha⁻¹ contained 20.6 nmol P (mg starch)⁻¹. The greenhouse-grown plants were cultured in inert media and received nutrient solutions containing 0, 3, 12 or 24 mg P l⁻¹, respectively. Plants grown with no P-fertilizer produced tuber starch with phosphorylation levels reduced to approximately 30% of the level found in plants grown with an ample supply of P. Thus, the level of starch phosphorylation can be modified by limiting the P-supply through the root system. Application of foliar P-fertilizer to the greenhouse-grown plants had no significant effect on the phosphorylation of the tuber starch.