Sample pooling for real-time PCR detection and virulence determination of the footrot pathogen <Emphasis Type="Italic">Dichelobacter nodosus</Emphasis>

Dichelobacter nodosus is the principal cause of ovine footrot and strain virulence is an important factor in disease severity. Therefore, detection and virulence determination of D. nodosus is important for proper diagnosis of the disease. Today this is possible by real-time PCR analysis. Analysis of large numbers of samples is costly and laborious; therefore, pooling of individual samples is common in surveillance programs. However, pooling can reduce the sensitivity of the method. The aim of this study was to develop a pooling method for real-time PCR analysis that would allow sensitive detection and simultaneous virulence determination of D. nodosus. A total of 225 sheep from 17 flocks were sampled using ESwabs within the Swedish Footrot Control Program in 2014. Samples were first analysed individually and then in pools of five by real-time PCR assays targeting the 16S rRNA and aprV2/B2 genes of D. nodosus. Each pool consisted of four negative and one positive D. nodosus samples with varying amounts of the bacterium. In the individual analysis, 61 (27.1%) samples were positive in the 16S rRNA and the aprV2/B2 PCR assays and 164 (72.9%) samples were negative. All samples positive in the aprV2/B2 PCR-assay were of aprB2 variant. The pooled analysis showed that all 41 pools were also positive for D. nodosus 16S rRNA and the aprB2 variant. The diagnostic sensitivity for pooled and individual samples was therefore similar. Our method includes concentration of the bacteria before DNA-extraction. This may account for the maintenance of diagnostic sensitivity. Diagnostic sensitivity in the real-time PCR assays of the pooled samples were comparable to the sensitivity obtained for individually analysed samples. Even sub-clinical infections were able to be detected in the pooled PCR samples which is important for control of the disease. This method may therefore be implemented in footrot control programs where it can replace analysis of individual samples.     

Insulin dysregulation

Abnormalities of insulin metabolism include hyperinsulinaemia and insulin resistance, and these problems are collectively referred to as insulin dysregulation in this review. Insulin dysregulation is a key component of equine metabolic syndrome: a collection of endocrine and metabolic abnormalities associated with the development of laminitis in horses, ponies and donkeys. Insulin dysregulation can also accompany prematurity and systemic illness in foals. Causes of insulin resistance are discussed, including pathological conditions of obesity, systemic inflammation and pituitary pars intermedia dysfunction, as well as the physiological responses to stress and pregnancy. Most of the discussion of insulin dysregulation to date has focused on insulin resistance, but there is increasing interest in hyperinsulinaemia itself and insulin responses to feeding. An oral sugar test or in‐feed oral glucose tolerance test can be performed to assess insulin responses to dietary carbohydrates, and these tests are now recommended for use in clinical practice. Incretin hormones are likely to play an important role in postprandial hyperinsulinaemia and are the subject of current research. Insulin resistance exacerbates hyperinsulinaemia, and insulin sensitivity can be measured by performing a combined glucose‐insulin test or i.v. insulin tolerance test. In both of these tests, exogenous insulin is administered and the rate of glucose uptake into tissues measured. Diagnosis and management of hyperinsulinaemia is recommended to reduce the risk of laminitis. The term insulin dysregulation is introduced here to refer collectively to excessive insulin responses to sugars, fasting hyperinsulinaemia and insulin resistance, which are all components of equine metabolic syndrome.     

Oral eosinophilic granulomas in tigers (Panthera tigris)--a collection of 16 cases.

Oral eosinophilic granulomas were diagnosed in 16 tigers (Panthera tigris). All lesions were located on the hard or soft palate and typically consisted of flat or slightly raised circular ulcers. Histologic features of these lesions were essentially identical to those seen in oral eosinophilic granulomas of domestic cats and dogs. No clinical signs were noted in eight cases, though various degrees of inappetence, excessive salivation, and dysphagia were noted in the other eight tigers. Six cases were not treated. Treatment for the remaining 10 cases centered on corticosteroids and additional treatments included surgical removal, cryotherapy, antibiotics, and chlorpheniramine. Treatment with corticosteroids did appear to be effective in some cases, though lesions would worsen after cessation of therapy and no cases were cured. In addition, three cases developed complications possibly related to this corticosteroid therapy. The etiology of these lesions remains unknown, though an underlying allergic condition is likely.     

Integrated approach for improving small scale market oriented dairy systems in Pakistan: participatory rural appraisal and economic opportunity survey

Livestock production is an integral part of the rain-fed and irrigated agriculture system in Pakistan. Animal production is closely interlinked with the cropping systems and play a crucial role in the rural economy. Participatory rural appraisals and economic opportunity surveys were conducted in two ecological zones (irrigated and rain-fed) and two dairy production systems (peri-urban and mixed livestock). Major constraints in animal health, nutrition and reproduction were identified and interventions were suggested to overcome these difficulties. The economic opportunity survey revealed that maximum opportunity to enhance farmers’ income is to increase milk production per day per animal, which can be accomplished through coordinated improvements in nutrition, reproduction and genetics.     

Effects of oral administration of levothyroxine sodium on concentrations of plasma lipids, concentration and composition of very-low-density lipoproteins, and glucose dynamics in healthy adult mares

OBJECTIVE: To evaluate glucose and lipid metabolism in healthy adult horses administered levothyroxine sodium (L-T4). ANIMALS: 12 healthy adult mares. PROCEDURE: 8 horses received an incrementally increasing dosage of L-T4 (24, 48, 72, or 96 mg of L-T4/d) for weeks 1 to 8. Each dose was provide between 7 AM and 8 AM in the morning grain meal for 2 weeks. Four additional horses remained untreated. Serum concentrations of nonesterified fatty acids, triglyceride (TG), total cholesterol (TC), and very-low-density lipoprotein (VLDL) were measured and composition of VLDL examined in samples obtained between 8 AM and 9 AM at weeks 0, 2, 4, 6, and 8. Glucose dynamics were assessed by use of a combined IV glucose-insulin tolerance test (IVGITT) conducted before and at the end of the 8-week treatment period. Data for each combined IVGITT were interpreted by use of the minimal model. RESULTS: Plasma TG, TC, and VLDL concentrations significantly decreased over time in treated horses. At the completion of the 8-week treatment period, mean plasma VLDL concentration was 46% of the mean value for week 0 in treated horses. Insulin sensitivity significantly increased (> 2-fold) in treated horses, but glucose effectiveness and net insulin response were not affected. Levothyroxine sodium significantly increased the rate of insulin disposal. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of L-T4 decreases blood lipid concentrations, improves insulin sensitivity, and increases insulin disposal in horses. Levothyroxine sodium may have potential as a treatment for horses with reduced insulin sensitivity.     

Incidence and molecular characterisation of lumpy skin disease virus in Zimbabwe using the P32 gene

Between January, 2013 and December, 2014, there was a lumpy skin disease (LSD) outbreak that affected cattle in different localities of Zimbabwe. The outbreak resulted in severe economic losses to the livestock industry. A retrospective study was conducted by examining stored veterinary records of the LSD outbreak at the Central Veterinary Laboratory (CVL) in Harare, Zimbabwe. Over the 2-year period, a total of 10,038 cases and 880 deaths (8.77 %) were recorded. LSD was reported from all regions of the country, with the highest incidence occurring in Mashonaland West (30.95 %) and Midlands province (14.59 %). The frequency of reported outbreaks was highest in March and April, with the lowest reported cases occurring in November. A total of 25 representative specimens (skin biopsies) were collected from nodular skin lesions of infected cattle, and after viral DNA isolation, the P32 gene was successfully amplified, by using PCR, in 88 % (22/25) of all assayed specimens. Out of the 22 samples that showed amplification, 16 (73 %) were selected for DNA sequencing, and from these, 13 sequences were submitted to GenBank and assigned accession numbers: KX033494, KX033495, KX033496, KX033497, KXO33498, KX033499, KX033500, KX033501, KX033502, KX033503, KX033504, KX033505 and KX033506. Phylogenetic analyses of the 13 sequences was done by using MEGA 7 and showed that the viruses formed two major clusters implying that at least two strains of LSDV are in circulation in Zimbabwe. This study provides the first report on the incidence and molecular characterisation of LSDV in Zimbabwe.     

Nutraceutic effect of free condensed tannins of <Emphasis Type="Italic">Lysiloma acapulcensis</Emphasis> (Kunth) benth on parasite infection and performance of Pelibuey sheep

Forty-five Pelibuey sheep were experimentally infested with nematodes to evaluate the effect of three free condensed tannin (FCT) levels of Lysiloma acapulcensis on fecal egg counts (FECs), packed cell volumes (PCV), ocular mucosa colors (OMC), average daily gain (ADG), and adult nematode count. Five treatments were used: 12.5, 25.0, and 37.5 mg of FCT kg^?1 of body weight (BW); sterile water (control); and ivermectine (0.22 mg kg^?1 of BW) as chemical group. The data were processed through repeated measurement analysis. Even though the three FCT doses decreased (P?<?0.05) the FEC, the highest reduction was obtained with 37.5 mg kg^?1 of BW. No differences were observed in PCV and OMC. Higher ADG (P?<?0.05) was observed with 37.5 mg kg^?1 of BW of FCT. The count of adult nematodes (females and males) in the higher dose of FCT was similar to chemical treatment. Dose of 37.5 mg kg^?1 of BW decreased the parasite infection and improved the lamb performance. Therefore, this dose could be used as a nutraceutic product in sheep production.     

Apparent metabolizable energy value of whole date palm (<Emphasis Type="Italic">Phoenix dactylifera</Emphasis> L.) and its possible use as a feedstuff for aged laying hens

This study was conducted to evaluate the feeding value of whole date palm (WDP) with and without enzyme supplementation on aged laying hen’s performance. Apparent metabolizable energy value of WDP was determined by the total collection method using ten adult leghorn cockerels. WDP was substituted with a corn-soybean meal basal diet at 40% level, and then the metabolizable energy of this experimental diet and basal diet was determined. After that, a total number of 256 Bovanz 95-week-old hens were randomly allocated into eight groups consisting of four replicates of eight birds each, based on a 4 × 2 factorial arrangement of the treatments. Eight iso-caloric and iso-nitrogenous experimental diets including one corn-soybean meal-based control diet and two, three, and four corn-soybean meal-based diets included 70, 140, and 210 g/kg ground WDP, respectively. Each of the diets supplemented with two levels of an enzyme (0.0 and 0.07 g/kg Natozim Plus). There was no significant difference in feed intake, feed conversion ratio, egg production, egg mass, eggshell thickness, and Haugh unit among the treatments (P > 0.05). However, yolk color score significantly decreased as dietary WDP level increased. The serum biochemical metabolites were not significantly affected by WDP and enzyme supplementation (P > 0.05). There were no significant differences in the relative weight of different organs except for abdominal fat. Our findings show that using WDP up to 21% of the diet was more economic and had no adverse effect on productive performance and serum metabolites of laying hens. However, WDP had an adverse effect on yolk color which can be ameliorated by carotenoid supplementation.     

An assessment of benzimidazole resistance against caprine nematodes in Central India

Current status of resistance to benzimidazole (BZ) group of anthelmintic drugs against caprine nematodes in Central India at Amanala goat farm, Jabalpur, Madhya Pradesh (M. P.), was systematically investigated using faecal egg count reduction (FECR) test and egg hatch test (EHT). Besides, allele-specific PCR (AS-PCR) was deployed to ascertain the susceptible genotype (alleles) especially of the Haemonchus contortus. Randomly selected 30 goats, irrespective of age and sex, were divided into three groups of 10 each, to serve as treated and untreated controls. It was ensured that the animals were not administered with an anthelmintic drug for the past 3 months prior to undertaking the study, and faecal egg counts were estimated. FECR test evidenced fenbendazole resistance by partial elimination (24.90%) copro-egg counts in the treated group of animals vis-à-vis controls with a lower confidence interval of ?26%. Further, EHT revealed ED-50 value of 0.335 μg of thiabendazole/ml, confirming benzimidazole resistance in the animals of that farm. AS-PCR showed that 62% of H. contortus larvae were homozygous resistant (rr), 24% heterozygous (rS) and 14% homozygous susceptible (SS). The genotypic frequencies of three genotypes (rr, rS and SS) were significantly (P < 0.01) different. The prevalence of benzimidazole resistance allele (r) was also significantly (P < 0.01) higher (74%) as compared to susceptible allele (S) (26%). The resistance to benzimidazole has been discussed while emphasizing improved managemental practices designed to reduce exposure of the goat population to parasites, minimize frequency of anthelmintic use at optimum dose and rotational use of different chemical groups of medicines with different mode of action, so as to overcome and combat the upcoming problem in the field.