Evaluating the Influence of Different Recommended Dietary Levels of Cu and Zn on Finishing Pigs

This study was conducted to evaluate the effects of dietary supplementation of different recommended levels of Cu and Zn on antioxidant capacity, tissue mineral status, minerals excretion, meat quality, digestive enzyme activity, and metal transporters in finishing pigs. A total of 120 pigs (with an average initial body weight (BW) of 70.0 ± 2.1 kg) were randomly divided into four treatments: (1) basal diet without added Cu or Zn (control), (2) basal diet+35 mg cupreous N-carbamylglutamate chelate (NCG-Cu) +150 mg zinc-methionine chelate (Zn-Met) (AC), (3) basal diet + 3.0 mg of NCG-Cu + 43 mg Zn-Met (CN), and (4) basal diet + 3.5 mg NCG-Cu + 50 mg Zn-Met (NRC100). Pig growth performance was not affected by the level of Cu or Zn. Among the four treatments, the AC treatment had the highest concentration (P < 0.05) of glutathione peroxidase (GSH-Px). Pigs fed the AC diet had the highest (P < 0.05) liver Zn, fecal Cu, and fecal Zn among the four treatments. The protein levels of trypsin and aminopeptidase N (APN) in the intestinal mucosa showed their highest levels (P < 0.05) in the NRC100 and AC treatments. The mRNA levels of trypsinogen and APN were significantly up-regulated (P < 0.05) in the AC, CN, and NRC100 treatments compared with the control. The mRNA levels for the Zn transporter genes SLC30A1 (ZnT1) and SLC30A2 (ZnT2) were significantly up-regulated (P < 0.05) in the AC treatment, and the mRNA levels for SLC39A4 (ZIP4) and metallothionein 1 (MT) in the AC, CN, and NRC100 treatments were significantly up-regulated (P < 0.05) compared with the control. Meat quality were not affected (P > 0.05) by the different recommended levels of Cu and Zn. These results indicated that the supplemental Cu and Zn levels routinely used in AC diets in Chinese commercial feed enterprises should be reduced.

Case Report: Long-Term Survival of a Dog With Chronic Lymphocytic Leukemia Treated With Chlorambucil, Prednisolone, and Imatinib

A 7-year-old castrated male Poodle dog presented with chronic progressive lymphocytosis. Hematologic and peripheral blood smear findings included remarkable lymphocytosis with well-differentiated small lymphocytes. Cytology of bone marrow aspirate showed hypercellular integrity with infiltration of small mature lymphocytes, accounting for 45% of all nucleated cells. Flow cytometry of blood and marrow samples revealed neoplastic lymphocytes predominantly expressing the CD21 molecule. B-cell chronic lymphocytic leukemia (CLL) was diagnosed on an immunophenotypic analysis. Administrations of prednisolone and chlorambucil were initiated and the response was unremarkable. Therefore, additional treatment with imatinib was provided, which resolved the hematologic abnormalities associated with CLL. Flow cytometry after ~1 year of treatment showed normalization of the count of lymphocytes positive for CD21 and resolved hematologic lymphocytosis. The dog was followed-up for 2 years, and there were no severe adverse effects. This case indicates that imatinib may be a good option as an adjunctive therapy with prednisolone and chlorambucil treatment for CLL in dogs without treatment response.

Effects of Nerve Growth Factor-β From Bull Seminal Plasma on Steroidogenesis and Angiogenic Markers of the Bovine Pre-ovulatory Follicle Wall Cell Culture

Nerve growth factor-β (NGF) is critical for ovulation in the mammalian ovary and is luteotrophic when administered systemically to camelids and cattle. This study aimed to assess the direct effects of purified bovine NGF on steroidogenesis and angiogenic markers in the bovine pre-ovulatory follicle. Holstein heifers (n = 2) were synchronized with a standard protocol, and heifers with a preovulatory follicle (≥ 12 mm) had the ovary containing the dominant follicle removed via colpotomy. Pre-ovulatory follicles were dissected into 24 pieces containing theca and granulosa cells that were randomly allocated into culture media supplemented with either purified bovine NGF (100 ng/mL) or untreated (control) for 72 h. The supernatant media was harvested for quantification of progesterone, testosterone, and estradiol concentrations, whereas explants were subjected to mRNA analyses to assess expression of steroidogenic and angiogenic markers. Treatment of follicle wall pieces with NGF upregulated gene expression of steroidogenic enzyme HDS17B (P = 0.04) and increased testosterone production (P < 0.01). However, NGF treatment did not alter production of progesterone (P = 0.81) or estradiol (P = 0.14). Consistently, gene expression of steroidogenic enzymes responsible for producing these hormones (STAR, CYP11A1, HSD3B, CYP17A1, CYP19A1) were unaffected by NGF treatment (P ≥ 0.31). Treatment with NGF downregulated gene expression of the angiogenic enzyme FGF2 (P = 0.02) but did not alter PGES (P = 0.63), VEGFA (P = 0.44), and ESR1 (P = 0.77). Collectively, these results demonstrate that NGF from seminal plasma may interact directly on the theca and granulosa cells of the bovine pre-ovulatory follicle to stimulate testosterone production, which may be secondary to theca cell proliferation. Additionally, decreased FGF2 expression in NGF-treated follicle wall cells suggests hastened onset of follicle wall cellular remodeling that occurs during early luteal development.

The Synergistic Effects of Combined Use of Mentha longifolia, Thymus carmanicus, and Trachyspermum copticum on Growth Performance, Feed Utilization, and Expression of Key Immune Genes in Rainbow Trout (Oncorhynchus mykiss)

Medicinal plants exhibit remarkable positive effects on different aspects of fish physiology. This study aimed to evaluate the possible impact of a combination of plants (Mentha longifolia, Thymus carmanicus, and Trachyspermum copticum) on growth performance, immune responses and key immune gene expression of rainbow trout. For this purpose, four diets were designed, including zero, 0.25, 0.5, and 1% of a mixture of plants per kg of diet, representing dietary treatments of control, T1, T2, and T3, respectively. Two hundred forty fish (weighing 23.11 ± 0.57 g) were fed 3% of body weight twice a day for 45 days. The results showed that growth parameters of weight gain (except for T1) and FCR were significantly improved in fish receiving all levels of plants, with T3 showing the best growth results. Digestive enzymes activities were notably increased in T1 and T2 compared to the control. Stress biomarkers (glucose and cortisol) were significantly decreased in T1 and T2, while T3 was not significantly different from the control. Immunological responses were significantly improved in T2, while T1 andT3 did not show a statistical difference in terms of lysozyme activity. Catalase activity was noticeably decreased in T1, although superoxide dismutase and malondialdehyde were highest in T2. Immune-related genes were significantly up-regulated in T3 compared to other treatments. Also, antioxidant enzyme coding genes were strongly up-regulated in T2 and T3. Overall, the present results suggest that 1% inclusion of the mixture of M. longifolia, T. carmanicus, and T. copticum (T2) can be used to improve the growth and immunity of rainbow trout.

Association Study of the M132L Single Nucleotide Polymorphism With Susceptibility to Chronic Wasting Disease in Korean Elk: A Meta-Analysis

Chronic wasting disease (CWD) is a deleterious brain proteinopathy caused by a pathogenic form of prion protein (PrPSc), which is converted from a benign form of prion protein (PrPC) encoded by the prion protein gene (PRNP). In elk, the M132L single nucleotide polymorphism (SNP) of the PRNP gene likely plays a pivotal role in susceptibility to CWD. However, the association of the M132L SNP with susceptibility to CWD has not been evaluated in Korean elk to date. To estimate the association of the M132L SNP with susceptibility to CWD in Korean elk, we investigated the genotype and allele frequencies of the M132L SNP by amplicon sequencing and performed association analysis between CWD-positive and CWD-negative elk. In addition, we performed a meta-analysis to evaluate the association between the M132L SNP and susceptibility to CWD in quantitatively synthesized elk populations. Furthermore, we estimated the effect of the M132L SNP on elk PrP using in silico programs, including PolyPhen-2, PROVEAN, AMYCO and Swiss-PdbViewer. We did not identify a significant association between the M132L SNP of PRNP and susceptibility to CWD in Korean elk. The meta-analysis also did not identify a strong association between the M132L SNP of PRNP and susceptibility to CWD in quantitatively synthesized elk populations. Furthermore, we did not observe significant changes in structure, amyloid propensity or electrostatic potential based on the M132L SNP in elk PrP. To the best of our knowledge, this was the first report of an association analysis and meta-analysis in Korean elk and quantitatively synthesized elk populations, respectively.

What Will Happen to This Dog? A Qualitative Analysis of Rehoming Organisations' Pre-adoption Dog Behaviour Screening Policies and Procedures

Rehoming organisations often undertake some type of behaviour evaluation to determine dogs' suitability for rehoming and/or the type of suitable home. Assessments can carry considerable weight in determining dogs' fates. Although evaluation of the validity and reliability of any test is important, a more fundamental consideration is if the nature of the information sought and the weight given to this in organisations' decision making is of more than anecdotal value. Therefore, this study's aim was to conduct a qualitative analysis of organisations' pre-adoption dog behaviour screenings and potential justifications, comparing this with the available scientific evidence. A written enquiry was sent electronically to rehoming organisations in the UK and US from February 2016-July 2017. Of 73 respondents, the majority conducted assessments for all dogs. Using a thematic analysis, nine themes and 71 sub-themes emerged concerning the types of information respondents aim to gather from assessments. The majority of respondents used, at least partially, pass/fail scoring, i.e., certain outcomes would lead to dogs being deemed unadoptable. Forty-one sub-themes and one theme were identified as potentially leading to a dog being deemed unadoptable. The evidence base for these factors was identified from the scientific literature relating to: increased risk for relinquishment, impact on a dog's quality of life, and human safety risk. Evidence supported 10 factors: “aggression towards people”, “aggression towards cats or other animals”, “aggression towards dogs”, “biting or snapping”, “resource guarding”, “activity level or exercise needs”, “destructiveness”, “housetrained”, “fearfulness”, and “knowledge of basic commands and/or general training”. Of those, seven were associated only with relinquishment risk, two (“resource guarding”, “knowledge of basic commands”) with human safety risk, and one (“fearfulness”) with both. Thus, for >85% of characteristics organisations deemed important for dogs' adoptability, scientific evidence to support this is lacking. More research is needed to investigate the value of behaviour assessments, especially concerning the assessment of factors that could pose a public safety risk. However, given the current lack of scientific support for many decisions regarding dogs' rehoming suitability and recognised pressure on resources, it is suggested that organisations should focus on pre-adoption adopter education and post-adoption support.

Hematological and Serum Biochemical Changes and Their Prognostic Value in Horses Spontaneously Poisoned by Crotalaria spectabilis

Determining the prognosis of poisoning by plants containing pyrrolizidine alkaloids is usually challenging. This study aimed to identify important prognostic parameters that can determine the severity of spontaneous poisoning by Crotalaria spectabilis in horses. Blood samples from 42 horses spontaneously poisoned by oats contaminated with C. spectabilis seeds were evaluated. Complete blood counts (CBC) and serum biochemical tests [urea, creatinine, total protein, albumin, total and direct bilirubin concentrations, aspartate aminotransferase (AST), γ-glutamyl transferase (GGT), and creatine kinase (CK) activities] were performed. Horses were followed up for 12 months to determine the long-term survival rate; after 12 months, they were divided into two groups: survivors (n = 30) and non-survivors (n = 12). Horses spontaneously poisoned with C. spectabilis had higher levels of urea, globulin, bilirubin (total, direct, and indirect), AST, GGT, and CK than the reference values. Non-survivor horses showed significantly higher (p < 0.05) values of hemoglobin, GGT, and direct bilirubin than the survivor horses. Horses with serum GGT activity higher than 95 U/l had 14.0 times the risk of death compared to animals showing activities equal to or lower than this value, whereas horses with serum direct bilirubin concentration higher than 0.6 mg/dl (10.26 μmol/L) had 5.78 times the risk of death compared to the others. In summary, serum GGT activity and direct bilirubin concentration may be useful prognostic indicators for assessing the severity of C. spectabilis-poisoned horses.

Long-Term Follow-Up of Restorations of Equine Cheek Teeth Infundibula (2006–2017)

Background:Caries of the infundibula of equine cheek teeth can lead to significant dental disease including increased attritional wear, pulpar and apical disease, secondary sinusitis, and dental fracture. Restorations of cavities of equine cheek teeth infundibula have been performed since 1889. Recent advances in dental materials, instrumentation, and techniques have facilitated the use of dental restoration techniques by equine veterinary practitioners. No studies to date have demonstrated the safety or efficacy of restorations of equine cheek teeth infundibula.Objectives:To assess the long-term results of restorations of equine cheek teeth affected by infundibular caries, to report on the safety of the procedure, and to give guidelines for future restorative therapies.Study Design:Retrospective analysis of results of clinical and oroscopic examination of horses that underwent infundibular restoration procedures between 2006 and 2017.Methods:A total of 223 infundibula in 185 maxillary cheek teeth in 92 horses were restored using a variety of dental materials including glass ionomer cement, flowable and compactible resin composites. The time between restoration and re-examination was recorded along with findings of clinical signs in the interim, restorative material loss, and any further pathological changes of the teeth including caries progression, fracture, or apical disease. Follow-up examinations were performed over two study periods 2006–2012 and in 2017.Results:Over the full study period, 99% of treated horses available for follow-up examinations had no adverse clinical signs or developed any abnormalities of restored teeth observable on oroscopic examination. Of horses re-examined, 83% of restorations were shown to have minimal or no loss of the restoration material, with occlusal surface wear visibly comparable to other adjacent maxillary teeth. Statistical analysis showed success of the procedure was related to the restorative material used, the restoration technique, and the caries grade present at the time of restoration (grade 2 is more successful than grade 3).Main Limitations:There are no case controls in this study and therefore it is not clear if restoration of equine infundibula is a consistently beneficial procedure, or at which grade of caries progression restorations should be performed for optimum benefit. The procedures were not re-examined at consistent regular times creating some difficulties in standardizing results. Re-examinations of treated horses did not consistently include radiography or computed tomography and therefore some apical changes may have occurred in treated teeth without visual oroscopic or external clinical signs.Conclusion:Restoration of equine infundibula using materials developed for human dentistry including flowable resin composites is a safe and long-lasting procedure and appears to prevent the development of further pathological changes including apical infection and dental fracture.

Acute Succinate Administration Increases Oxidative Phosphorylation and Skeletal Muscle Explosive Strength via SUCNR1

Endurance training and explosive strength training, with different contraction protein and energy metabolism adaptation in skeletal muscle, are both beneficial for physical function and quality of life. Our previous study found that chronic succinate feeding enhanced the endurance exercise of mice by inducing skeletal muscle fiber-type transformation. The purpose of this study is to investigate the effect of acute succinate administration on skeletal muscle explosive strength and its potential mechanism. Succinate was injected to mature mice to explore the acute effect of succinate on skeletal muscle explosive strength. And C2C12 cells were used to verify the short-term effect of succinate on oxidative phosphorylation. Then the cells interfered with succinate receptor 1 (SUCNR1) siRNA, and the SUCNR1-GKO mouse model was used for verifying the role of SUCNR1 in succinate-induced muscle metabolism and expression and explosive strength. The results showed that acute injection of succinate remarkably improved the explosive strength in mice and also decreased the ratio of nicotinamide adenine dinucleotide (NADH) to NAD+ and increased the mitochondrial complex enzyme activity and creatine kinase (CK) activity in skeletal muscle tissue. Similarly, treatment of C2C12 cells with succinate revealed that succinate significantly enhanced oxidative phosphorylation with increased adenosine triphosphate (ATP) content, CK, and the activities of mitochondrial complex I and complex II, but with decreased lactate content, reactive oxygen species (ROS) content, and NADH/NAD+ ratio. Moreover, the succinate's effects on oxidative phosphorylation were blocked in SUCNR1-KD cells and SUCNR1-KO mice. In addition, succinate-induced explosive strength was also abolished by SUCNR1 knockout. All the results indicate that acute succinate administration increases oxidative phosphorylation and skeletal muscle explosive strength in a SUCNR1-dependent manner.

Cryptosporidium and Giardia in Livestock in Tigray, Northern Ethiopia and Associated Risk Factors for Infection: A Cross-Sectional Study

The occurrence and species/genotypes of Cryptosporidium and Giardia duodenalis infecting young livestock in selected districts of Tigray, Ethiopia were investigated, along with risks associated with infection. A total of 757 faecal samples were collected from calves, lambs, and goat kids from four rural districts in Tigray, and also from calves in periurban Mekelle, Tigray's main city, and analysed for Cryptosporidium oocysts and Giardia cysts. Farmers answered questionnaires regarding potential risk factors at sample collection. Immunofluorescent antibody staining was used for parasite detection, and PCR at selected genes and sequencing of positive samples was used for molecular characterisation. The occurrence of Cryptosporidium infection was 10, 9, and 4% in calves, lambs, and goat kids, respectively; equivalent figures for Giardia infection were 39, 32, and 21%. Molecular characterisation of Cryptosporidium isolates revealed C. ubiquitum, subtype XIIa in all three host species; C. ryanae in calves and goat kids; C. andersoni and C. bovis were identified only in calves, and C. xiaoi was identified in lambs. For Giardia, Assemblage E predominated in all host species, but among calf isolates we also identified a few potentially zoonotic genotypes (assemblages A (AI) and Assemblage B). Periparturient care was shown to be a particularly relevant risk factor for infection, and infections were less likely to occur under extensive management systems. Our major findings were widespread occurrence of both parasites in livestock, and the apparent lack of the most common zoonotic species. Our results are discussed in relation to other relevant studies. As our study was conducted in Tigray, further investigation in different settings in Ethiopia could provide relevant information on transmission and zoonotic potential. In addition, given the dependency on healthy animals for the livelihoods of the population of Tigray, investigation of the effect of these common parasites on livestock productivity is important.