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Objective To determine whether surgical removal of urachal anomalies improves the outcomes of dogs with recurrent lower urinary tract disease (LUTD) and bacterial urinary tract infection (BUTI). Study design Retrospective study. Animals Thirty-three dogs with urachal anomalies and recurrent LUTD or BUTI. Methods Medical records of dogs with LUTD or BUTI and a diagnosis of urachal anomaly treated by partial cystectomy were reviewed. A minimum follow-up of 9 months was required for inclusion. Results Median age at onset of clinical signs was 12 months (range, 1 month to 10 years). Urachal anomalies were detected with histopathology in 20 of 28 (71%) dogs. At a median follow-up of 22 months (range, 9-114), 21 of 28 (64%) dogs were free of signs of LUTD. Nine (27%) dogs exhibited reduced signs of LUTD; in three (9%) dogs, no clinical improvement was observed. Among the 25 dogs with confirmed preoperative BUTI, 22 clinically improved with surgery. Conclusion Partial cystectomy reduced the long-term severity of clinical signs and risk of recurrence of LUTD or BUTI in dogs with confirmed or suspected urachal anomalies. Clinical significance Partial cystectomy should be considered as an adjunct to the treatment of LUTD and BUTI in dogs.

BACKGROUND: Chronic hepatitis (CH) in dogs is common and has the tendency to progress to liver cirrhosis (LC). Circulating microRNAs might have the potential as markers for disease progression. OBJECTIVES: To investigate whether concentration of specific microRNAs in serum correlate with the stage and grade of CH in Labrador Retrievers. ANIMALS: Twenty-two Labrador Retrievers with histological CH (n = 8), LC (n = 7), and normal liver (NL, n = 7). METHODS: In this retrospective study, serum concentrations of miR-122, miR-29a, miR-133a, miR-181b, and miR-17-5p were measured by quantitative real-time PCR and evaluated using univariate linear regression in dogs. A multivariate model was fit including the grade of hepatitis and the stage of fibrosis. RESULTS: Of the 5 microRNAs, only circulating miR-122 and miR-29a were significantly associated with the grade of hepatitis and the stage of fibrosis. A positive correlation was identified between the grade of hepatitis with miR-122 (rs = 0.79, P

Summary Hepatic steatosis is a highly prevalent liver disease, yet research is hampered by the lack of tractable cellular and animal models. Steatosis also occurs in cats, where it can cause severe hepatic failure. Previous studies demonstrate the potential of liver organoids for modeling genetic diseases. To examine the possibility of using organoids to model steatosis, we established a long-term feline liver organoid culture with adult liver stem cell characteristics and differentiation potential toward hepatocyte-like cells. Next, organoids from mouse, human, dog, and cat liver were provided with fatty acids. Lipid accumulation was observed in all organoids and interestingly, feline liver organoids accumulated more lipid droplets than human organoids. Finally, we demonstrate effects of interference with β-oxidation on lipid accumulation in feline liver organoids. In conclusion, feline liver organoids can be successfully cultured and display a predisposition for lipid accumulation, making them an interesting model in hepatic steatosis research., Highlights • Feline organoids can be cultured from fresh and frozen liver and from needle biopsy • A feline-specific liver culture medium allows organoid expansion up to 32 passages • When fed fatty acids, feline liver organoids store more lipids than human organoids • Feline liver organoids can be used to model hepatic steatosis, In this study Kruitwagen and colleagues establish and characterize a feline liver organoid culture, which has adult stem cell properties and can be differentiated toward hepatocyte-like cells. They propose liver organoids as a tool to model hepatic steatosis and show that feline liver organoids accumulate more lipids than human organoids when provided with excess fatty acids.

International audience; Harbour porpoises (Phocoena phocoena) are the most prevalent cetaceans in the North Sea. The fecal viral flora of 21 harbour porpoises stranded along the Dutch coastline was analyzed by a metagenomics approach. Sequences of a novel cetacean mastadenovirus, designated harbour porpoise adenovirus 1 (HpAdV-1), were detected. The sequence of a 23-kbp genomic region, spanning the conserved late region, was determined using primer walking. Phylogenetic analysis indicated that HpAdV-1 is most closely related to bottlenose dolphin adenovirus and clusters with Cetartiodactyla adenoviruses. The prevalence of HpAdV-1 was low (2.6%) based on targeted PCR-screening of the intestinal contents of 151 harbour porpoises stranded between 2010 and 2013.

8658860258318000Recently, genetic alterations in the genes encoding succinate dehydrogenase subunit B and D (SDHB and SDHD) were identified in pet dogs that presented with spontaneously arising pheochromocytomas (PCC) and paragangliomas (PGL, together PPGL), suggesting dogs might be an interesting comparative model for the study of human PPGL. To study whether canine PPGL resembled human PPGL, we investigated a series of 50 canine PPGLs by immunohistochemistry to determine the expression of synaptophysin (SYP), tyrosine hydroxylase (TH) and succinate dehydrogenase subunit A (SDHA) and B (SDHB). In parallel, 25 canine PPGLs were screened for mutations in SDHB and SDHD by Sanger sequencing. To detect large chromosomal alterations, single nucleotide polymorphism (SNP) arrays were performed for 11 PPGLs, including cases for which fresh frozen tissue was available. The immunohistochemical markers stained positive in the majority of canine PPGLs. Genetic screening of the canine tumors revealed the previously described variants in four cases, SDHB p.Arg38Gln (n = 1) and SDHD p.Lys122Arg (n = 3). Furthermore, the SNP arrays revealed large chromosomal alterations of which the loss of chromosome 5, partly homologous to human chromosome 1p and chromosome 11, was the most frequent finding (100% of the six cases with chromosomal alterations). In conclusion, canine and human PPGLs show similar genomic alterations, suggestive of common interspecies PPGL-related pathways.

Objective To report the long‐term outcome of nine dogs treated for caudal cervical spondylomyelopathy (CCSM) with surgical spinal fusion. Study design Short case series. Animals Nine large‐breed dogs. Methods Medical records of dogs treated for disc‐associated CCSM (2013‐2016) were reviewed. The surgery objective was spinal distraction by implantation of a SynCage and fixation with two Unilock plates. Follow‐up included the Helsinki pain score questionnaire, neurological grading, radiography, computed tomography (CT), and micro‐CT (μCT) with subsequent histopathology (two dogs). Results Clinical follow‐up was obtained between 9 and 51 months (27.4 ± 13.4 months). The Helsinki pain score and neurological Griffith score improved (P

No standardized treatment guidelines are reported in veterinary medicine for dogs with large pituitary corticotroph neoplasms causing neurological signs, and such dogs usually have a short overall survival. When these dogs undergo pituitary surgery and the tumor regrows there are few reports of subsequent treatments. A 7 yr old male Maltese diagnosed with pituitary-dependent hypercortisolism developed seizures in conjunction with a large pituitary corticotroph adenoma and underwent transsphenoidal hypophysectomy. After 3 yr of clinical remission, hypercortisolism recurred, and trilostane therapy was initiated. One year later, the dog developed new neurological signs and computed tomography revealed regrowth of a large pituitary mass that was then treated with radiation therapy. The dog lived disease-free for 3 more yr. At postmortem examination, a more aggressive pituitary neoplasm than the one examined at the time of surgery was found, which is suggestive of malignant transformation into a carcinoma despite the absence of convincing metastasis.

Four outbreaks of leukoencephalomyelopathy in colonies of SPF cats on a long‐term diet of irradiated dry cat food were observed in the Netherlands between 1989 and 2001. As a primary defect in myelin formation was suspected to be the cause of the disease and myelin consists mainly of lipids and their fatty acids, we investigated the fatty acid composition of the white matter of the spinal cord of affected and control cats and of irradiated and non‐irradiated food. The irradiated food had low levels of alpha‐linolenic acid compared to linoleic acid as well as a high total omega‐6:omega‐3 ratio of 7:1 in the irradiated and of 2:1 in the non‐irradiated food. The white matter of the spinal cord showed low levels of linoleic acid and absence of alpha‐linolenic acid in affected cats as well as absence of lignoceric and nervonic acid in both affected and control cats. These abnormalities in fatty acid composition of the white matter of the spinal cord may reflect an increased need for alpha‐linolenic acid as a substrate for longer chain omega‐3 fatty acids to compose myelin and thus indicate a particular species sensitivity to dietary deficiency in omega‐3 polyunsaturated fatty acids, particularly alpha‐linolenic acid in cats. Our findings indicate that abnormalities in fatty acid metabolism in myelin play an essential role in the pathogenesis of this acquired form of leukoencephalomyelopathy in cats.

In 2017, endocarditis caused by Streptococcus equi subspecies zooepidemicus was diagnosed in a man in the Netherlands who had daily contact with horses. Whole-genome sequencing of isolates from the man and his horses confirmed the same clone, indicating horse-to-human transmission. Systematic reporting of all zoonotic cases would help with risk assessment.

This retrospective study determined disease free survival (DFS) and progression free survival (PFS) in chemo-naïve dogs with multicentric or cranial mediastinal high-grade T-cell lymphoma, treated with a first-line CCNU-L(-chlorambucil)-CHOP protocol. Of thirteen dogs with multicentric lymphoma, 92.3% achieved a complete remission (CR), and the median DFS and PFS was 317 and 256 days, respectively. Three dogs had cranial mediastinal lymphoma, and achieved a CR with a median DFS and PFS of 978 and 1007 days, respectively. The oneand two-year DFS/PFS probability estimate for dogs with multicentric lymphoma was 0.50/0.46 and 0.42/0.38, respectively, for dogs with cranial mediastinal lymphoma 0.67/0.67. Neutropenia and thrombocytopenia were reported in 52.9% and 50% of the dogs, respectively, while 56.3% experienced grade 1-4 nephrotoxicity, hypothesized to be lomustine-induced. It was concluded that, compared to historical data, the currently described first-line CCNU-L(-chlorambucil)- CHOP protocol could benefit the survival of dogs with multicentric or cranial mediastinal highgrade T-cell lymphoma.

Previous reports have shown that reproductive tract score (RTS) can predict reproduction outcomes in seasonally bred beef heifers, although the accuracy can vary. Some ultrasonographic measures of the female reproductive tract and pelvis area have also been associated with reproductive outcome in young heifers. The objectives of this study were to determine which transrectal ultrasound or pelvis measures taken at a single examination are independent predictors of reproductive failure and whether the RTS system can be optimized with this information. In this observational study, year-old beef heifers (n = 488) in 2 birth cohorts were followed from just before the first breeding until confirmation of pregnancy. A single pre-breeding examination included body condition score, RTS, ultrasound measures of the reproductive tract (length and diameter of the left and right ovaries, presence and diameter of a CL, largest follicle diameter and left uterus horn diameter) and transverse and vertical diameters of the pelvis. Additional farm records including dam parity, sire, birth weight and birth date, weaning weight, weaning date, prebreeding body weight, AI dates, and semen used were available. Breeding consisted of 50 days of AI, followed 5 to 7 days later by a 42-day bull breeding period. Pregnancy failure was defined as the failure to become pregnant after the AI and bull breeding periods, while anestrus was defined as the failure to be detected in estrus during the 50-day AI period. From the prebreeding data and farm records, independent predictors of pregnancy failure and anestrus were identified using stepwise reduction in multiple logistic regression models. Age at the onset of breeding was the only consistent independent predictor of pregnancy failure and anestrus in both cohorts of this study (P < 0.05). Body condition score, uterus horn diameter, absence of a CL, largest follicle of less than 13 mm, and pelvis area (PA) were the prebreeding examination variables that remained in prognostic models (P < 0.1). Combining either the model based on the 3 remaining ultrasound measures or RTS with PA provided more accurate prognostic models for pregnancy failure and anestrus than using RTS alone (P < 0.05). It is concluded that ultrasound measures have prognostic value for pregnancy failure in restricted bred yearling heifers as a result of their association with anestrus, and that smaller PA has additional prognostic value for poorly performing heifers.

The occurrence of the zoonotic protozoan parasite Toxoplasma gondii in marine mammals remains a poorly understood phenomenon. In this study, samples from 589 marine mammal species and 34 European otters (Lutra lutra), stranded on the coasts of Scotland, Belgium, France, The Netherlands and Germany, were tested for the presence of T. gondii. Brain samples were analysed by polymerase chain reaction (PCR) for detection of parasite DNA. Blood and muscle fluid samples were tested for specific antibodies using a modified agglutination test (MAT), a commercial multi-species enzyme-linked immunosorbent assay (ELISA) and an immunofluorescence assay (IFA). Out of 193 animals tested by PCR, only two harbour porpoise (Phocoena phocoena) cerebrum samples, obtained from animals stranded on the Dutch coast, tested positive. The serological results showed a wide variation depending on the test used. Using a cut-off value of 1/40 dilution in MAT, 141 out of 292 animals (41%) were positive. Using IFA, 30 out of 244 tested samples (12%) were positive at a 1/50 dilution. The commercial ELISA yielded 7% positives with a cut-off of the sample-to-positive (S/P) ratio ≥ 50; and 12% when the cut-off was set at S/P ratio ≥ 20. The high number of positives in MAT may be an overestimation due to the high degree of haemolysis of the samples and/or the presence of lipids. The ELISA results could be an underestimation due to the use of a multispecies conjugate. Our results confirm the presence of T. gondii in marine mammals in The Netherlands and show exposure to the parasite in both the North Sea and the Eastern Atlantic Ocean. We also highlight the limitations of the tests used to diagnose T. gondii in stranded marine mammals.

Effective management of natural resources involves a multidisciplinary perspective to address complex issues in data poor-environments. With mobile species that do not conform to human-defined borders a cross-boundary approach is essential. There is a continuing concern of ecological sustainability of marine environments, which demands monitoring of ecosystem indicators. Such indicators are increasingly derived from monitoring sentinel species. Harbour porpoises (Phocoena phocoena) are included as indicator species in several national and international agreements. Increasing exposure to anthropogenic stressors may impact harbour porpoise populations. To investigate these risks, a better understanding of threats and their effect is required. This study aimed to identify current knowledge gaps, to predict future pressures or threats, and to define useful conservation indicators to facilitate future research on harbour porpoises in the North Sea, through expert elicitation gained in a two-round Delphi approach. The three most important knowledge gaps addressed were bycatch, population dynamics, and the cumulative effects of multiple stressors. Bycatch was predicted as the highest concern for porpoises in the next 20 years, followed by chemical and noise pollution, respectively. A list of essential indicators aiming to increase understanding of harbour porpoises’ health status was established and studying causes of death, distribution, abundance, habitat use and diet composition were scored as most relevant. These results should guide research focus and management objectives of harbour porpoise populations and the study design could be translated to serve managers in other geographical areas aiming to identify knowledge gaps and defining research priorities for other wildlife species.

Hypophosphatemia is a common finding in periparturient and anorectic cattle. Although the clinical relevance of hypophosphatemia in cattle is uncertain, it has been empirically associated with persistent recumbency, specifically in periparturient dairy cows. The objective of the present study was to determine if transient dietary phosphorus (P) deprivation over a course of 5 wk, by feeding an approximately 40% P-deficient ration to lactating dairy cows, would result in altered muscle function or muscle P metabolism severe enough to present a risk for animal health and well-being. In addition, we wanted to determine the association between the plasma phosphate concentration ([Pi]) and muscle tissue P content to assess to what extent intracellular P deprivation of muscle cells could be extrapolated from subnormal plasma [Pi]. Ten healthy multiparous, mid-lactating dairy cows received a ration with a P content of 0.18% over a period of 5 wk. Following the P-deprivation phase, the same ration supplemented with P to obtain a dietary P content of 0.43% was fed for 2 wk. Blood and urine samples were collected regularly and muscle biopsies were obtained repeatedly to determine the P content in muscle tissue. Function of skeletal and heart muscles was evaluated by electrocardiography and electromyography conducted repeatedly throughout the study. Feeding the P-deficient ration resulted in the rapid development of marked hypophosphatemia. The lowest plasma [Pi] were measured after 9 d of P depletion and were, on average, 60% below predepletion values. Plasma [Pi] increased thereafter, despite ongoing dietary P depletion. None of the animals developed clinical signs commonly associated with hypophosphatemia or any other health issues. Urine analysis revealed increasing renal calcium, pyridinoline, and hydroxypyridinoline excretion with ongoing P deprivation. Biochemical muscle tissue analysis showed that dietary P depletion and hypophosphatemia were not associated with a decline in muscle tissue P content. Electromyographic examination revealed increased occurrence of pathological spontaneous activity in striated muscles after 2 wk of dietary P depletion in several cows, which could be suggestive of neuromuscular membrane instability. No effect on heart muscle activity was identified electrocardiographically. These results suggest that counter-regulatory mechanisms were sufficient to maintain normal muscle tissue P content during transient and moderate P deprivation. Muscle function was not grossly affected, although the increased occurrence of pathological spontaneous activity suggests that subclinical neuropathy or myopathy, or both, may have occurred with ongoing P deprivation. The results presented here indicate that plasma [Pi] is unsuitable for assessing muscle tissue P content in cattle.

OBJECTIVE: To determine whether polyxylon bandage contact lenses influence healing time and ocular comfort in the management of spontaneous chronic corneal epithelial defects in dogs. METHODS: Twenty dogs with spontaneous chronic corneal epithelial defects were included. All dogs were treated by debridement under topical anaesthesia at the first presentation. Ten dogs were assigned to the study group (application of a polyxylon bandage contact lens), and the remaining ten served as a control group. The healing time and ocular (dis)comfort were evaluated by assessment of the clinical findings and an owner-based questionnaire. All dogs received the same topical and systemic medication. RESULTS: Healing time for dogs in the study population was significantly shorter (mean 14±0 days) than for dogs in the control group (mean 36±17 days; P=0·005). The spontaneous chronic corneal epithelial defects had completely healed at the first recheck in all dogs with a polyxylon bandage contact lens. The duration of blepharospasm following debridement was significantly shorter in the study population (mean 4±4 days) than in the control group (mean 30±20 days; P=0·001). CLINICAL SIGNIFICANCE: The use of polyxylon bandage contact lenses is beneficial in the management of spontaneous chronic corneal epithelial defects.

This study aimed at determining the incidence, distribution, risk factors, and causal serotypes of foot and mouth disease (FMD) outbreaks in Ethiopia based on 5 years of retrospective outbreak data (September 2007 until August 2012). District level outbreak data were collected from 115 randomly selected districts using a questionnaire administered to district animal health officers. The national incidence of FMD outbreaks during the study period was 1.45 outbreaks per five district years. Outbreaks were geographically widespread affecting all major regional states in the country and were more frequent in the central, southern, and southeastern parts of the country. Neither long-term nor seasonal trends were observed in the incidence of outbreaks. A mixed effects logistic regression analysis revealed that the type of production system (market oriented system versus subsistence systems), presence of a major livestock market and/or route, and adjacency to a national parks or wildlife sanctuary were found to be associated with increased risk of outbreaks in the districts. FMD virus serotypes O, A, SAT 2, and SAT 1 were identified as the causal serotypes of the outbreaks during the study period. Whereas O was the dominant serotype, SAT 2 was the serotype that showed increase in relative frequency of occurrence. The estimated incidence of outbreaks is useful in assessing the economic impacts of the disease, and the identified risk factors provide important knowledge to target a progressive FMD control policy for Ethiopia.

BACKGROUND: Hypercortisolism is a common endocrine disorder in dogs, caused by a cortisol-secreting adrenocortical tumor (AT) in approximately 15% of cases. In adrenocortical carcinomas of humans, activation of the phosphatidylinositol 3 kinase (PI3K) signaling pathway by insulin-like growth factor (IGF) signaling represents a promising therapeutic target. OBJECTIVES: To investigate the involvement of PI3K signaling in the pathogenesis of ATs in dogs and to identify pathway components that may hold promise as future therapeutic targets or as prognostic markers. ANIMALS: Analyses were performed on 36 canine cortisol-secreting ATs (11 adenomas and 25 carcinomas) and 15 normal adrenal glands of dogs. METHODS: mRNA expression analysis was performed for PI3K target genes, PI3K inhibitor phosphatase and tensin homolog (PTEN), IGFs, IGF receptors, IGF binding proteins and epidermal growth factor receptors. Mutation analysis was performed on genes encoding PTEN and PI3K catalytic subunit (PIK3CA). RESULTS: Target gene expression indicated PI3K activation in carcinomas, but not in adenomas. No amino acid-changing mutations were detected in PTEN or PIK3CA and no significant alterations in IGF-II or IGFR1 expression were detected. In carcinomas, ERBB2 expression tended to be higher than in normal adrenal glands, and higher expression of inhibitor of differentiation 1 and 2 (ID1 and ID2) was detected in carcinomas with recurrence within 2.5 years after adrenalectomy. CONCLUSIONS AND CLINICAL IMPORTANCE: Based on these results, ERBB2 might be a promising therapeutic target in ATs in dogs, whereas ID1 and 2 might be valuable as prognostic markers and therapeutic targets.

In vitro gas production studies are routinely used to assess the metabolic capacity of intestinal microbiota to ferment dietary fibre sources. The faecal inocula used during the in vitro gas production procedure are most often obtained from animals adapted to a certain diet. The present study was designed to assess whether 19 days of adaptation to a diet are sufficient for faecal inocula of pigs to reach a stable microbial composition and activity as determined by in vitro gas production. Eighteen multiparous sows were allotted to one of two treatments for three weeks: a diet high in fibre (H) or a diet low in fibre (L). After this 3-week period, the H group was transferred to the low fibre diet (HL-treatment) while the L group was transferred to the diet high in fibre (LH-treatment). Faecal samples were collected from each sow at 1, 4, 7, 10, 13, 16 and 19 days after the diet change and prepared as inoculum used for incubation with three contrasting fermentable substrates: oligofructose, soya pectin and cellulose. In addition, inocula were characterised using a phylogenetic microarray targeting the pig gastrointestinal tract microbiota. Time after diet change had an effect (P

Several parameters for improving volatile metabolite analysis using headspace gas chromatography-mass spectrometry (GC-MS) analysis of volatile metabolites were evaluated in the framework of identification of mastitis-causing pathogens. Previous research showed that the results of such volatile metabolites analysis were comparable with those based on bacteriological culturing. The aim of this study was to evaluate the effect of several method changes on the applicability and potential implementation of this method in practice. The use of a relatively polar column is advantageous, resulting in a faster and less complex chromatographic setup with a higher resolving power yielding higher-quality data. Before volatile metabolite analysis is applied, a minimum incubation of 8 h is advised, as reducing incubation time leads to less reliable pathogen identification. Application of GC-MS remained favorable compared with regular gas chromatography. The complexity and cost of a GC-MS system are such that this limits the application of the method in practice for identification of mastitis-causing pathogens.

Objective—To evaluate serum concentrations of biochemical markers and survival time in dogs with protein-losing enteropathy (PLE). Design—Prospective study. Animals—29 dogs with PLE and 18 dogs with food-responsive diarrhea (FRD). Procedures—Data regarding serum concentrations of various biochemical markers at the initial evaluation were available for 18 of the 29 dogs with PLE and compared with findings for dogs with FRD. Correlations between biochemical marker concentrations and survival time (interval between time of initial evaluation and death or euthanasia) for dogs with PLE were evaluated. Results—Serum C-reactive protein concentration was high in 13 of 18 dogs with PLE and in 2 of 18 dogs with FRD. Serum concentration of canine pancreatic lipase immunoreactivity was high in 3 dogs with PLE but within the reference interval in all dogs with FRD. Serum α1-proteinase inhibitor concentration was less than the lower reference limit in 9 dogs with PLE and 1 dog with FRD. Compared with findings in dogs with FRD, values of those 3 variables in dogs with PLE were significantly different. Serum calprotectin (measured by radioimmunoassay and ELISA) and S100A12 concentrations were high but did not differ significantly between groups. Seventeen of the 29 dogs with PLE were euthanized owing to this disease; median survival time was 67 days (range, 2 to 2,551 days). Conclusions and Clinical Relevance—Serum C-reactive protein, canine pancreatic lipase immunoreactivity, and α1-proteinase inhibitor concentrations differed significantly between dogs with PLE and FRD. Most initial biomarker concentrations were not predictive of survival time in dogs with PLE.

Harbour porpoises are often found to be infected by endoparasites in several organs including the lungs and stomach as well as the heart, liver and ears. Nevertheless there is still little knowledge about the impact, ecology, transmission, and virulence of these parasitic infections. Here, we profile the presence of parasites in 4 frequently infected organs (lungs, stomach, liver and ears) in relation to biological parameters of harbour porpoises stranded along the Dutch coastline between December 2008 and December 2013. We found that parasites were common, with prevalence of 68% in lungs, 74.4% in ears, 26% in stomach and 23.5% in liver. We used generalised linear models to further quantify parasite presence in relation to biological data gathered during necropsy (sex, body length and nutritive condition). Body length (used as a proxy for age) was significant in explaining parasite presence for all organs with increasing probability of having the parasite with increasing body length. For the parasitic infections in the ears and stomach the nutritive condition was an additional significant factor, with a higher probability of parasite presence in porpoises in a poorer nutritive condition. The results of this study can be used as a baseline for assessing parasite presence in harbour porpoises and are a first step towards linking parasite infections to basic biological data gathered during necropsy.

Degus (Octodon degus) are prone to develop dental disease with deleterious health effects. The two studies reported here aimed to determine the prevalence of dental disorders in degus and to identify and evaluate diagnostic tools for determination of prognosis of these disorders. In study A, health data from 225 degus at AAP, Rescue Center for Exotic Animals in the Netherlands, were collated and the prevalence of dental disorders and differences in sex and age at clinical onset of symptoms associated with dental disorders were described. The prevalence was 34.7 per cent and higher (P

We examined consistency of the relationship between intramammary infection (IMI) and somatic cell score (SCS) across several classes of cow, herd, and sampling time variables. Microbial cultures of composite milk samples were performed by New York Quality Milk Production Services from 1992 to 2004. SCS was from the most recent Dairy Herd Improvement test before IMI sampling. Records were analyzed from 79,308 cows in 1,124 commercial dairy herds representing a broad range of production systems. Three binary dependent variables were presence or absence of contagious IMI, environmental IMI, and all IMI. Independent variables in the initial models were SCS, SCS2, lactation number, days in milk, sample day milk yield, use of coliform mastitis vaccine, participant type (required by regulation or voluntary), production system (type of housing, milking system, and herd size), season of sampling, year of sampling, and herd; also the initial models included interactions of SCS and SCS2 with other independent variables, except herd and milk yield. Interaction terms characterize differences in the IMI-SCS relationship across classes of the independent variables. Models were derived using the Glimmix macro in SAS (SAS Institute Inc., Cary, NC) with a logistic link function and employing backward elimination. The final model for each dependent variable included all significant independent variables and interactions. Simplified models omitted SCS2 and all interactions with SCS. Interactions of SCS with days in milk, use of coliform mastitis vaccine, participant type, season, and year were not significant in any of the models. Interaction of SCS with production system was significant for the all IMI model, whereas interaction of SCS with lactation number was significant for the environmental and all IMI models. Each 1 point increase in SCS (or doubling of somatic cell count) was associated with a 2.3, 5.5%, and 9.1% increase in prevalence of contagious, environmental, and all IMI, respectively. Empirical receiver operator characteristic curves and areas under the curve were derived for final and simplified models. The areas under the curve for simplified and final models within each type of IMI differed by 0.009 or less. We concluded that the relationship of IMI with SCS was generally stable over time and consistent across seasons, production systems, and cow factors.

REASONS FOR PERFORMING STUDY: Hypoventilation or apnoea, caused by the induction of general anaesthesia, may cause hypoxaemia. Pre-oxygenation may lengthen the period before this happens. No scientific studies are published on pre-oxygenation in equine anaesthesia. OBJECTIVES: To determine whether supplementation of oxygen at a flow rate of 15 litres per minute for 3 minutes via a nasal cannula before induction of general anaesthesia is effective in elevating the arterial partial pressure of oxygen (PaO2 ) directly after induction. STUDY DESIGN: Randomised, prospective clinical trial. METHODS: Eighteen adult horses, American Society of Anesthesiologists physical status 1 or 2, undergoing elective anaesthesia were randomly allocated to one of 2 groups. The first group (control group) received no oxygen supplementation before induction of general anaesthesia, whereas the second group (oxygen group) did. All horses were anaesthetised with intravenous detomidine, butorphanol, ketamine, midazolam and isoflurane. Directly after induction (T = 0) and 30 min later (T = 30) an arterial blood sample was taken for blood gas analysis. At T = 30 an estimate of intrapulmonary shunt fraction (Qs/Qt) was calculated. RESULTS: At T = 0 arterial partial pressure of oxygen (PaO2 ) was significantly higher in the oxygen group compared to the control group (11.0 ± 2.6 kPa versus 7.4 ± 1.6 kPa; mean ± s.d., p = 0.005), at T = 30 differences were not statistically significant. Partial pressure of carbon dioxide (PaCO2 ) and Qs/Qt did not differ between the groups. CONCLUSIONS: Supplementing oxygen by a nasal cannula before induction of general anaesthesia in horses is feasible and does effectively elevate the PaO2 immediately after induction. Future research is needed to determine whether supplementation of oxygen before induction of general anaesthesia in horses will affect outcomes. This article is protected by copyright. All rights reserved.

Background The use of antimicrobials in food animals and the emergence of antimicrobial resistance are global concerns. Ceftiofur is the only third-generation cephalosporin labeled for veterinary use in the USA, and it is the drug of choice in the majority of dairy farms for the treatment of mastitis. Here, we use next-generation sequencing to describe longitudinal changes that occur in the milk microbiome before, during, and after infection and treatment with ceftiofur. Twelve animals were intramammary challenged with Escherichia coli in one quarter and randomly allocated to receive intramammary treatment with ceftiofur (5d) or untreated controls. Serial samples were collected from −72 to 216 h relative to challenge from the challenged quarter, an ipsilateral quarter assigned to the same treatment group, and from a third quarter that did not undergo intervention. Results Infection with E. coli dramatically impacted microbial diversity. Ceftiofur significantly decreased LogCFUs but had no significant effect on the milk microbiome, rate of pathogen clearance, or somatic cell count. At the end of the study, the microbial profile of infected quarters was indistinguishable from pre-challenge samples in both treated and untreated animals. Intramammary infusion with ceftiofur did not alter the healthy milk (i.e., milk devoid of clots or serous appearance and collected from a mammary gland that shows no clinical signs of mastitis) microbiome. Conclusions Our results indicate that the mammary gland harbors a resilient microbiome, capable of reestablishing itself after experimental infection with E. coli independent of antimicrobial treatment. Electronic supplementary material The online version of this article (doi:10.1186/s40168-017-0291-5) contains supplementary material, which is available to authorized users.

The effect of browse species tannins (using polyethylene glycol, PEG 6000) on in vitro gas production (GP), methane (CH 4 ), ammonia (NH 3 ), volatile fatty acids (VFA) and in vitro organic matter digestibility (IVOMD) were studied. Approximately 0.5 g of air-dried A. etbaica, C. farinosa, C. tomentosa, D. angustifolia, D. cinerea, E. racemosa, M. angolensis, M. senegalensis, R. natalensis and S. singueana leaves were used as substrates in an automated in vitro system. Proanthocyanidin (PA) contents were quantified using the modified HCl-butanol method and the PA composition analysed by ultra performance liquid chromatography using a diode array detector coupled with electrospray ionization mass spectrometry (UPLC-DAD-ESI–MS/MS). Substrates were inoculated for 72 h in buffered rumen fluid (2:1, v/v), pooled from three goats fed a grass silage and concentrate diet, on three separate occasions. Each substrate was incubated in triplicate with or without inclusion of PEG 6000 and GP measured. During incubation, head space gas samples were taken at 0, 3, 6, 9, 12, 24, 30, 48, 54, and 72 h and analysed for CH 4 . Volatile fatty acids, NH 3 and IVOMD were determined after 72 h of incubation. Data from three runs were averaged for analysis. Addition of PEG increased (P < 0.0001) GP, CH 4 , NH 3 and total VFA indicating that PA were mainly involved in reducing methanogenesis but also digestibility. Prodelphinidins were found to be the major PA affecting fermentation. Also the contribution of quercetin, myricetin and kaempferol derivatives in CH 4 reduction were evident. Changes in the molar proportions of VFA with PEG addition indicated that PA affected fermentation pathways. The absence of PEG effect on IVOMD was due to artefacts from the tannin-PEG complexes interfering with the incubation residue measurement. Overall, the effect of tannin-containing browse on in vitro fermentation characteristics was mainly due to PA with the possible minor effects of other phenolic and non-phenolic compounds.

A cortisol-secreting adrenocortical tumour (ACT) is the cause of naturally occurring canine hypercortisolism in approximately 15-20% of cases. The differentiation between an adrenocortical adenoma and carcinoma is usually based on histopathology. However, histopathological parameters have never been linked to the dogs' survival. Moreover, in human medicine the interobserver variability of some histopathological parameters that are used for ACTs is high. The objective of this study was to establish a reliable and easy-to-use histopathological scoring system for cortisol-secreting ACTs that can assess the prognosis of dogs after adrenalectomy. Cortisol-secreting ACTs of 50 dogs, collected between 2002 and 2015, were included in this study. Twenty histopathological features were assessed by one veterinary pathologist and one resident in veterinary pathology. In addition, the Ki67 proliferation index was assessed by two observers. Only parameters with intra- and interobserver agreement scores (intraclass correlation or Cohen's kappa coefficient) of ≥0.40 were included in survival analyses. Use of multivariate forward stepwise regression analysis with associated hazard ratios led us to a scoring system which we call the Utrecht score: the Ki67 proliferation index, +4 if more than 33% of the tumour cells have clear/vacuolated cytoplasm, and +3 if necrosis is present. Using cut-off values of 6 and 11, we could distinguish three groups that had significantly shorter survival times with increasing Utrecht scores. We conclude that the Utrecht score can be used to assess the prognosis of dogs with cortisol-secreting ACTs after adrenalectomy, which can help to select high-risk dogs that might benefit from adjuvant treatment or additional monitoring. This article is protected by copyright. All rights reserved.

During intervertebral disc (IVD) maturation, the main cell type shifts from notochordal cells (NCs) to chondrocyte-like cells (CLCs). NCs secrete factors with regenerative potential, making them an interesting focus for regenerative treatments. During initial development, these strategies preferably employ non-human donors due to easy availability of their NC-rich nucleus pulposus (NP) tissue. To increase the success of translating these strategies for clinical application, this study aimed to delineate whether NC-secreted factors of different species have a regenerative effect on human CLCs. Human, canine and porcine NC-rich NP tissue and NC-conditioned medium (NCCM) were analysed biochemically and histologically. Human CLC micro-aggregates from degenerated IVDs were cultured in human, canine or porcine NCCM. Collagen, glycosaminoglycan (GAG) and DNA content was determined and histology was performed. Canine and porcine NPs were richer in NCs than human NPs. Human NPs contained the highest collagen content, whereas the DNA and GAG content of canine NPs was significantly higher than that of human or porcine NPs. NCCM from all species significantly increased the DNA and GAG content of the human CLC micro-aggregates. Porcine and canine NCCM were significantly more potent than human NCCM in inducing GAG deposition, whereas only human NCCM induced collagen type II production. Secreted factors from human, canine and porcine NC-rich NPs exerted regenerative effects on human CLCs, indicating a cross-species effect. Bioactive compound(s) are present in NCCM of different species that may reverse human IVD degeneration, supporting further research into strategies based on NC-technology employing canine or porcine models for their translation into humans.

Background Campylobacter fetus (C. fetus) can cause disease in both humans and animals. C. fetus has been divided into three subspecies: C. fetus subsp. fetus (Cff), C. fetus subsp. venerealis (Cfv) and C. fetus subsp. testudinum (Cft). Subspecies identification of mammal-associated C. fetus strains is crucial in the control of Bovine Genital Campylobacteriosis (BGC), a syndrome associated with Cfv. The prescribed methods for subspecies identification of the Cff and Cfv isolates are: tolerance to 1 % glycine and H2S production. Results In this study, we observed the deletion of a putative cysteine transporter in the Cfv strains, which are not able to produce H2S from L-cysteine. Phylogenetic reconstruction of the core genome single nucleotide polymorphisms (SNPs) within Cff and Cfv strains divided these strains into five different clades and showed that the Cfv clade and a Cff clade evolved from a single Cff ancestor. Conclusions Multiple C. fetus clades were observed, which were not consistent with the biochemical differentiation of the strains. This suggests the need for a closer evaluation of the current C. fetus subspecies differentiation, considering that the phenotypic differentiation is still applied in BGC control programs. Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-3058-7) contains supplementary material, which is available to authorized users.

Selective removal of the pituitary adenoma has not been advocated in dogs with pituitary-dependent hypercortisolism because the pituitary adenoma is usually not visualized on routine computed tomography (CT). Dynamic pituitary CT scanning is aimed at the detection of the pituitary flush and, indirectly, at the presence and position of the adenoma. The first aim of this retrospective study was to compare findings of a multiple slice dynamic scanning protocol with those of a single slice dynamic protocol using a single slice CT scanner. The second aim was to compare the CT findings with surgical findings, and surgical findings with histopathological findings. Computed tomography with single and multiple slice dynamic scanning protocols was performed in 86 dogs with pituitary-dependent hypercortisolism. Thirty dogs underwent transsphenoidal hypophysectomy and pituitary specimens were collected as tumor, normal, mixed and neurohypophyseal samples and processed for histology. The pituitary flush was not detected more frequent in multiple slice dynamic scanning series than in single slice dynamic scanning series. However, in non-enlarged pituitaries, the flush was seen significantly more frequently than in enlarged pituitaries. Prediction of the nature of the tissue during hypophysectomy by the surgeon was inconclusive. In conclusion, when using a single slice CT scanner, both single or multiple slice dynamic scanning protocols can be used for localization of the neurohypophyseal flush, and, indirectly, the adenoma. However, based on this study, the aim of surgery in dogs with pituitary-dependent hypercortisolism remains total adenohypophysectomy, and when the neurophypophysis is recognized, it may be left in situ.

Udder cleft dermatitis (UCD) is a skin lesion in dairy cows, most often located between anterior parts of the udder and abdomen, but also found between the front quarters. A few recent studies have investigated the prevalence of UCD, but relatively little is known about its pathogenesis, clinical course, and duration. Therefore, the aim of this study was to investigate the incidence and recovery of UCD on high-prevalence herds. Five Dutch dairy herds with a UCD prevalence of at least 6% were visited weekly for 19 wk, followed by visits every other week for 26 wk. During each visit, all dry and lactating cows were inspected for the presence of UCD signs. If a UCD case was detected, the affected skin was photographed and the photo was subsequently examined by a research assistant. Cows were then classified according to the appearance of the skin into 3 categories: healthy (no photo: no signs), mild (photo: affected skin but no wound), or severe (photo: open wound). The overall mean within-herd prevalence of UCD was 38% and the overall mean incidence was 1.94 UCD episodes per 100 cow-weeks at risk. Incidence of UCD was significantly higher in cows in third or higher parity and significantly increased with DIM. Median observed duration of UCD was 16 wk. The UCD recovery was 3 times more likely for mild than for severe lesions. The probability of moving from one category to another between 2 consecutive visits was very low, indicating that rapid changes in appearance did not occur. The observed incidence of UCD was rather low, and the relatively high prevalence in the selected herds was most likely due to the long duration of lesions rather than a high incidence of new UCD cases.

Increasing antibiotic resistance and ever stricter control on antibiotic use are a driving force to develop alternatives to antibiotics. One such strategy is the use of multifunctional Host Defense Peptides. Here we examined the protective effect of prophylactic treatment with the D analog of chicken cathelicidin-2 (D-CATH-2) against a respiratory E. coli infection. Chickens were treated with D-CATH-2 in ovo at day 18 of embryonic development or intramuscularly at days 1 and 4 after hatch. At 7 days of age, birds were challenged intratracheally with avian pathogenic E. coli. Protection was evaluated by recording mortality, morbidity (Mean Lesion Score) and bacterial swabs of air sacs at 7 days post-infection. In ovo D-CATH-2 treatment significantly reduced morbidity (63%) and respiratory bacterial load (>90%), while intramuscular treatment was less effective. D-CATH-2 increased the percentage of peripheral blood lymphocytes and heterophils by both administration routes. E. coli specific IgM levels were lower in in ovo treated animals compared to intramuscular D-CATH-2 treatment. In short, in ovo treatment with the Host Defense Peptide derived D-CATH-2 can partially protect chickens from E. coli infection, making this peptide an interesting starting point to develop alternatives to antibiotics for use in the poultry sector.

Toxoplasma gondii can cause disease in goats, but also has impact on human health through food-borne transmission. Our aims were to determine the seroprevalence of T. gondii infection in indoor-housed Dutch dairy goats and to identify the risk factors related to T. gondii seroprevalence. Fifty-two out of ninety approached farmers with indoor-kept goats (58%) participated by answering a standardized questionnaire and contributing 32 goat blood samples each. Serum samples were tested for T. gondii SAG1 antibodies by ELISA and results showed that the frequency distribution of the log10-transformed OD-values fitted well with a binary mixture of a shifted gamma and a shifted reflected gamma distribution. The overall animal seroprevalence was 13.3% (95% CI: 11.7–14.9%), and at least one seropositive animal was found on 61.5% (95% CI: 48.3–74.7%) of the farms. To evaluate potential risk factors on herd level, three modeling strategies (Poisson, negative binomial and zero-inflated) were compared. The negative binomial model fitted the data best with the number of cats (1–4 cats: IR: 2.6, 95% CI: 1.1–6.5; > = 5 cats:IR: 14.2, 95% CI: 3.9–51.1) and mean animal age (IR: 1.5, 95% CI: 1.1–2.1) related to herd positivity. In conclusion, the ELISA test was 100% sensitive and specific based on binary mixture analysis. T. gondii infection is prevalent in indoor housed Dutch dairy goats but at a lower overall animal level seroprevalence than outdoor farmed goats in other European countries, and cat exposure is an important risk factor.

Determining the presence of functional gonadal tissue in dogs can be challenging, especially in bitches during anestrus or not known to have been ovariectomized, or in male dogs with nonscrotal testes. Furthermore, in male dogs treated with deslorelin, a slow-release GnRH agonist implant for reversible chemical castration, the verification of complete downregulation of the hypothalamic-pituitary-gonadal (HPG) axis can be difficult, especially if pretreatment parameters such as the size of the testes or prostate gland are not available. The aims of this study were to validate an immunoradiometric assay for measurement of FSH in canine urine, to determine if the urinary FSH to creatinine ratio can be used to verify the neuter status in bitches and male dogs, as an alternative to the plasma FSH concentration, and to determine if downregulation of the HPG axis is achieved in male dogs during deslorelin treatment. Recovery of added canine FSH and serial dilutions of urine reported that the immunoradiometric assay measures urinary FSH concentration accurately and with high precision. Plasma FSH concentrations (the mean of two samples, taken 40??minutes apart) and the urinary FSH to creatinine ratio were determined before gonadectomy and 140??days (median, range 121???225??days) and 206??days (median, range 158???294??days) after gonadectomy of 13 bitches and five male dogs, respectively, and in 13 male dogs before and 132??days (median, range 117???174??days) after administration of a deslorelin implant. In both bitches and male dogs, the plasma FSH concentration and the urinary FSH to creatinine ratio were significantly higher after gonadectomy, with no overlapping of their ranges. Receiver operating characteristic analysis of the urinary FSH to creatinine ratio revealed a cut-off value of 2.9 in bitches and 6.5 in males to verify the presence or absence of functional gonadal tissue. In male dogs treated with deslorelin, the plasma FSH concentrations and urinary FSH to creatinine ratios were significantly lower after administration of the implant, but their ranges overlapped. We conclude that the urinary FSH to creatinine ratio can be used to verify the neuter status of bitches and male dogs. However, it cannot be used for the assessment of complete downregulation of the HPG axis after administration of a deslorelin implant. The urinary FSH to creatinine ratio is preferable over the plasma FSH concentration because it involves only one sample that can be collected relatively easy and noninvasively.

30 sperm whales (Physeter macrocephalus) stranded along the coasts of the North Sea between January and February 2016. The gastro-intestinal tracts of 22 of the carcasses were investigated. Marine debris including netting, ropes, foils, packaging material and a part of a car were found in nine of the 22 individuals. Here we provide details about the findings and consequences for the animals. While none of the items was responsible for the death of the animal, the findings demonstrate the high level of exposure to marine debris and associated risks for large predators, such as the sperm whale.

In the four years following the first detection of ranavirus (genus Ranavirus, family Iridoviridae) infection in Dutch wildlife in 2010, amphibian mortality events were investigated nationwide to detect, characterize and map ranaviruses in amphibians over time, and to establish the affected host species and the clinico-pathological presentation of the disease in these hosts. The ultimate goal was to obtain more insight into ranavirus disease emergence and ecological risk. In total 155 dead amphibians from 52 sites were submitted between 2011 and 2014, and examined using histopathology, immunohistochemistry, virus isolation and molecular genetic characterization. Ranavirus-associated amphibian mortality events occurred at 18 sites (35%), initially only in proximity of the 2010 index site. Specimens belonging to approximately half of the native amphibian species were infected, including the threatened Pelobates fuscus (spadefoot toad). Clustered massive outbreaks involving dead adult specimens and ranavirus genomic identity indicated that one common midwife toad virus (CMTV)-like ranavirus strain is emerging in provinces in the north of the Netherlands. Modelling based on the spatiotemporal pattern of spread showed a high probability that this emerging virus will continue to be detected at new sites (the discrete reproductive power of this outbreak is 0.35). Phylogenetically distinct CMTV-like ranaviruses were found in the south of the Netherlands more recently. In addition to showing that CMTV-like ranaviruses threaten wild amphibian populations not only in Spain but also in the Netherlands, the current spread and risk of establishment reiterate that understanding the underlying causes of CMTV-like ranavirus emergence requires international attention.

This review discusses the scope of using (quantitative) electromyography (EMG) in diagnosing myopathies and neuropathies in equine patients. In human medicine, many EMG methods are available for the diagnosis, pathophysiological description and evaluation, monitoring, or rehabilitation of patients, and some of these techniques have also been applied to horses. EMG results are usually combined with other neurophysiological data, ultrasound, histochemistry, biochemistry of muscle biopsies, and clinical signs in order to provide a complete picture of the condition and its clinical course. EMG technology is commonly used in human medicine and has been subject to constant development and refinement since its introduction in 1929, but the usefulness of the technique in equine medicine is not yet widely acknowledged. The possibilities and limitations of some EMG applications for equine use are discussed.

Background Acquired aortopulmonary fistulation is a rare condition in humans. It usually results as a late complication of a true or pseudoaneurysm of the thoracic aorta. It is most commonly associated with trauma or surgery, less commonly with atherosclerosis, inflammation, hypertension or Marfan’s syndrome. Aortopulmonary fistulation is also seen as a rare complication of acute aortic dissection. On rare occasions, acquired aortopulmonary fistulation is reported in aged patients without any of the above mentioned triggering factors. Thus, these cases should be considered as idiopathic aortopulmonary fistulation. Clearly, the pathogenesis of this condition is not yet completely understood. Friesian horses are highly inbred and are affected by several genetic conditions. Rupture of the thoracic aorta has a relatively high prevalence in Friesian horses and is often characterized by the formation of a pseudoaneurysm with subsequent fistulation into the pulmonary artery. Affected animals may survive for several weeks to months. Findings Here we performed vascular casting in three affected Friesian horses. In all three cases, an aortic rupture at the caudoventral side of the aorta was connected with a rupture of the main pulmonary artery just proximal to its bifurcation. Conclusions Affected Friesians show a consistent location and configuration of the aortic rupture site, very similar to the human condition and therefore could act as a spontaneous model to study this disease. Electronic supplementary material The online version of this article (doi:10.1186/s13104-016-2201-5) contains supplementary material, which is available to authorized users.

In the first weeks of life young chickens are highly susceptible to infectious diseases due to immaturity of the immune system. Little is known about the expression of host defense peptides (HDPs) during this period. In this study we examined the expression pattern of two chicken HDPs, the cathelicidin CATH-2 and the β-defensin AvBD9 by immunohistochemistry in a set of organs from embryonic day 12 until four weeks posthatch. AvBD9 was predominantly found in enteroendocrine cells throughout the intestine, the first report of in vivo HDP expression in this cell type, and showed stable expression levels during development. CATH-2 was exclusively found in heterophils which decreased after hatch in most of the examined organs including spleen, bursa and small intestine. In the lung CATH-2 expression was biphasic and peaked at the first day posthatch. In short, CATH-2 and AvBD9 appear to be expressed in cell types strategically located to respond to infectious stimuli, suggesting these peptides play a role in embryonic and early posthatch defense.

In in situ nylon bag technique, many feed evaluation systems use a washing machine method (WMM) to determine the washout (W) fraction and to wash the rumen incubated nylon bags. As this method has some disadvantages, an alternate modified method (MM) was recently introduced. The aim of this study was to determine and compare the W and non-washout (D+U) fractions of nitrogen (N) and/or starch of maize and grass silages, using the WMM and the MM. Ninety-nine maize silage and 99 grass silage samples were selected with a broad range in chemical composition. The results showed a large range in the W, soluble (S) and D+U fractions of N of maize and grass silages and the W, insoluble washout (W-S) and D+U fractions of starch of maize silages, determined by both methods, due to variation in their chemical composition. The values for N fractions of maize and grass silages obtained with both methods were found different (p < 0.001). Large differences (p < 0.001) were found in the D+U fraction of starch of maize silages which might be due to different methodological approaches, such as different rinsing procedures (washing vs. shaking), duration of rinsing (40 min vs. 60 min) and different solvents (water vs. buffer solution). The large differences (p < 0.001) in the W-S and D+U fractions of starch determined with both methods can led to different predicted values for the effective rumen starch degradability. In conclusion, the MM with one recommended shaking procedure, performed under identical and controlled experimental conditions, can give more reliable results compared to the WMM, using different washing programs and procedures.

Morphologically, canine soft-tissue sarcomas (STSs) resemble human STSs. In humans, proper classification of STSs is considered essential to improve insight in the biology of these tumors, and to optimize diagnosis and therapy. To date, there is a paucity of data published on the significance of detailed classification of STSs in the dog. We revised a cohort ( n = 110) of proliferative lesions obtained from a study in Golden Retrievers that were considered “soft tissue sarcoma, not otherwise specified or of uncertain subtype” in order to optimize the diagnoses of these lesions. The criteria according to the veterinary WHO classification, recent veterinary literature, and the WHO classification for humans were applied. Revision was initially based on morphologic characteristics of hematoxylin and eosin–stained histologic sections of the neoplasms. If considered necessary ( n = 76), additional immunohistochemistry was applied to aid characterization. The diagnosis of STS was confirmed in 75 neoplasms (68%). Of this group, diagnosis of a specific subtype of the STSs was possible in 58 neoplasms. Seven neoplasms had morphologic characteristics that were suggestive for sarcoma subtypes only described in the WHO classification for humans. Seventeen neoplasms remained “unclassified STSs.” Thirty-one lesions (28%) were diagnosed “neoplasm, not being STS.” Four lesions (4%) were considered nonneoplastic. Because incorrect classification of a tumor could lead to inappropriate therapeutic intervention and prognostication, the results of our study clearly illustrate the importance of revision and further diagnosis of “unclassified STSs” in dogs.