DNA methylation and targeted sequencing of methyltransferases family genes in canine acute myeloid leukaemia,modelling human myeloid leukaemia

Tumours shows aberrant DNA methylation patterns, being hypermethylated or hypomethylated compared with normal tissues. In human acute myeloid leukaemia (hAML) mutations in DNA methyltransferase (DNMT3A) are associated to a more aggressive tumour behaviour. As AML is lethal in dogs, we defined global DNA methylation content, and screened the C‐terminal domain of DNMT3 family of genes for sequence variants in 39 canine acute myeloid leukaemia (cAML) cases. A heterogeneous pattern of DNA methylation was found among cAML samples, with subsets of cases being hypermethylated or hypomethylated compared with healthy controls; four recurrent single nucleotide variations (SNVs) were found in DNMT3L gene. Although SNVs were not directly correlated to whole genome DNA methylation levels, all hypomethylated cAML cases were homozygous for the deleterious mutation at p.Arg222Trp. This study contributes to understand genetic modifications of cAML, leading up to studies that will elucidate the role of methylome alterations in the pathogenesis of AML in dogs.     

Paediatric HUS Cases Related to the Consumption of Raw Milk Sold by Vending Machines in Italy: Quantitative Risk Assessment Based on Escherichia coli O157 Official Controls over 7 years

A quantitative risk assessment (RA) was developed to estimate haemolytic‐uremic syndrome (HUS) cases in paediatric population associated with the consumption of raw milk sold in vending machines in Italy. The historical national evolution of raw milk consumption phenomenon since 2008, when consumer interest started to grow, and after 7 years of marketing adjustment, is outlined. Exposure assessment was based on the official Shiga toxin‐producing Escherichia coli O157:H7 (STEC) microbiological records of raw milk samples from vending machines monitored by the regional Veterinary Authorities from 2008 to 2014, microbial growth during storage, consumption frequency of raw milk, serving size, consumption preference and age of consumers. The differential risk considered milk handled under regulation conditions (4°C throughout all phases) and the worst time–temperature field handling conditions detected. In case of boiling milk before consumption, we assumed that the risk of HUS is fixed at zero. The model estimates clearly show that the public health significance of HUS cases due to raw milk STEC contamination depends on the current variability surrounding the risk profile of the food and the consumer behaviour has more impact than milk storage scenario. The estimated HUS cases predicted by our model are roughly in line with the effective STEC O157‐associated HUS cases notified in Italy only when the proportion of consumers not boiling milk before consumption is assumed to be 1%. Raw milk consumption remains a source of E. coli O157:H7 for humans, but its overall relevance is likely to have subsided and significant caution should be exerted for temporal, geographical and consumers behaviour analysis. Health education programmes and regulatory actions are required to educate people, primarily children, on other STEC sources.     

Effect of intrafragmentary bupivacaine (haematoma block) on analgesic requirements in dogs undergoing fracture repair

Objective

To test the efficacy of intraoperative intrafragmentary administration of bupivacaine (haematoma block) in controlling postoperative pain in dogs undergoing osteosynthesis of long-bone isolated diaphyseal fractures.

Vaginourethroplasty for Treatment of Urethral Obstruction in the Bitch

Vaginourethroplasty was performed in six bitches with infiltrative, obstructive urethral disease. The initial clinical signs included dysuria, hematuria, pollakiuria, and stranguria. Diagnosis was based on the clinical signs, plain radiography and contrast vagino-urethrogram examination, and urethral biopsy. Five dogs were diagnosed as having urethral neoplasia; the sixth dog had granulomatous urethritis. In all six dogs, up to 50% of the urethra and the urethral tubercle were resected to ensure adequate surgical margins. One dog was euthanatized 4 months after surgery for recurrence of a transitional cell carcinoma at the surgical site. Long-term resolution of the problem (minimum of 12 months) was achieved in the other five dogs. The frequency of serious complications, including urinary incontinence and ascending lower urinary tract infections, was low.     

Effects of porcine relaxin on induced parturition in beef heifers

Two-year-old crossbred beef heifers were used to test the effects of porcine relaxin (pRelaxin) alone, or in combination with dexamethasone, on the induction of parturition, the incidence of dystocia, and retained placentas. Effects of treatment on pelvic area, postpartum interval, milk production, colostrum quality, calf birth weight, calf vigor, and calf performance were also evaluated. On Day 275 of gestation, heifers from two fetal-sire groups were randomly assigned to one of four groups in a 2 × 2 factorial design and received: no treatment (controls, N = 19), 20 mg of dexamethasone intramuscularly (im) (n = 22), 5 mg of pRelaxin (3,000 U/mg) im (n = 19), or 20 mg of dexamethasone plus 5 mg of pRelaxin (n = 17). Length of gestation (in days) was less (P < 0.05) in heifers treated with dexamethasone (279.8 ± 1.0) than in controls (286.6 ± 0.9), but was not influenced (P > 0.05) by treatment with pRelaxin. The incidence of retained placentas in heifers treated only with dexamethasone (27.3%) was not reduced by concomitant treatment with pRelaxin (35.3%). Retained placentas were not observed in any control heifers and in only one heifer (5.2%) treated solely with pRelaxin. Ease of calving (1 = unassisted, 5 = abnormal presentation) was not influenced by treatment (P > 0.05), even though birth weights (in kilograms) of calves from heifers treated with dexamethasone (36.4 ± 0.8) were less (P < .01) than those of calves from nondexamethasone-treated heifers (39.2 ± 0.8). Dexamethasone tended to reduce (P < 0.07) calf vigor (1 = healthy and strong, 5 = dead on arrival; 1.48 ± 0.11 vs. 1.18 ± 0.11), but was not (P > 0.05) influenced by pRelaxin. The duration of the postpartum anestrous interval (73.1 ± 1.8 d across groups) and pelvic areas following treatment and parturition were not influenced (P > 0.05) by dexamethasone or pRelaxin. Although determinants of colostrum quality (P < 0.01) and quantity (P < 0.08) of milk produced were influenced by dexamethasone, adjusted 205-d weights of calves did not differ (P > 0.05) among groups. In conclusion, treatment with pRelaxin alone failed to induce parturition or, when combined with dexamethasone, to reduce the incidence of retained placentas.     

Performance of broiler chicks fed on alkali‐treated neem (Azadirachta indica) kernel cake as a protein supplement

1. An experiment was conducted to evaluate growth and nutrient efficiency of broiler chicks from 3 to 42 d fed on diets containing alkali‐treated neem (Azadirachta indica A. juss) kernel cake (NKC) as a protein supplement in place of peanut meal (PNM).

2. NKC was treated with sodium hydroxide at 10 (ANKC 1) or 20 g (ANKC 2)/kg and incorporated into the test diets at 135 or 300 g/kg to replace 50 (low—L) or 100 (High—H) % of the PNM protein of the reference diet.

3. Despite comparable retentions of dry matter and total carbohydrate on L‐ANKC 1 and 2, fibre on L‐and H‐ANKC 2 and nitrogen, calcium and acid detergent fibre on all experimental diets compared to the retentions of chicks on the reference diet, only the chicks fed L‐ANKG 2 were found to grow and utilise food as well as those on the reference diet.

4. The activities of serum alkaline phosphatase on H‐ANKC 1 and alanine amino transferase on all test diets were depressed (P < 0.05), but the activity of serum aspartate amino transferase, total erythrocyte count and concentration of blood haemoglobin and urea were similar in all chicks.

5. No significant differences were noticed in the qualitative and quantitative characteristics of the meat of chicks fed on the reference diet and on diets incorporating ANKC at the lower concentrations. Feeding ANKC protein did not impart any untoward taste as evaluated in pressure cooked meat by a semitrained panel on a 7 point Hedonic scale.

6. Except for duodenal and jejunal inflammation in chicks on both reference and test diets, all the vital organs were normal, ruling out any adverse affects caused by residual neem bitters.

7. Comparable performance and cost of chicks fed on the reference and L‐ANKC 2 diets, warrants the utilisation of hitherto wasted protein‐rich NKC after alkali treatment in broiler chick diets to spare peanut meal for human consumption in developing countries.     

Ongoing Activity of Toscana Virus Genotype A and West Nile Virus Lineage 1 Strains in Turkey: A Clinical and Field Survey

Toscana virus (TOSV), West Nile virus (WNV) and tickborne encephalitis virus (TBEV) are among major viral pathogens causing febrile disease and meningitis/encephalitis. The impact of these viruses was investigated at a referral centre in Ankara Province, Central Anatolia in 2012, where previous reports suggested virus circulation but with scarce information on clinical cases and vector activity. Serum and/or cerebrospinal fluid samples from 94 individuals were evaluated, in addition to field‐collected arthropod specimens that included 767 sandflies and 239 mosquitoes. Viral nucleic acids in clinical samples and arthropods were sought via specific and generic nested/real‐time PCRs, and antibody responses in clinical samples were investigated via commercial indirect immunofluorescence tests (IIFTs) and virus neutralization. A WNV antigen assay was also employed for mosquitoes. WNV neuroinvasive disease has been identified in a 63‐year‐old male via RNA detection, and the WNV strain was characterized as lineage 1. TOSV infections were diagnosed in six individuals (6.3%) via RNA or IgM detection. Partial sequences in a 23‐year‐old female, presented with fever and transient pancytopenia, were characterized as TOSV genotype A. Febrile disease with arthralgia and/or peripheral cranial nerve involvement was noted in cases with TOSV infections. Previous WNV and TOSV exposures have been observed in 5.3% and 2.1% of the subjects, respectively. No confirmed TBEV exposure could be identified. Morphological identification of the field‐collected mosquitoes revealed Culex pipiens sensu lato (74.4%), Anopheles maculipennis (20.9%), An. claviger (2.1%) and others. Sandfly species were determined as Phlebotomus papatasi (36.2%), P. halepensis (27.3%), P. major s. l. (19.3%), P. sergenti (8.9%), P. perfiliewi (4.4%), P. simici (2.6%) and others. Viral infections in arthropods could not be demonstrated. TOSV genotype A and WNV lineage 1 activity have been demonstrated as well as serologically proven exposure in patients. Presence of sandfly and mosquito species capable of virus transmission has also been revealed.     

A Large‐scale,Rapid Public Health Response to Rabies in an Organ Recipient and the Previously Undiagnosed Organ Donor

This article describes and contrasts the public health response to two human rabies cases: one organ recipient diagnosed within days of symptom onset and the transplant donor who was diagnosed 18 months post‐symptom onset. In response to an organ‐transplant‐related rabies case diagnosed in 2013, organ donor and recipient investigations were conducted by multiple public health agencies. Persons with potential exposure to infectious patient materials were assessed for rabies virus exposure. An exposure investigation was conducted to determine the source of the organ donor's infection. Over 100 persons from more than 20 agencies spent over 2700 h conducting contact investigations in healthcare, military and community settings. The 564 persons assessed include 417 healthcare workers [5.8% recommended for post‐exposure prophylaxis (PEP)], 96 community contacts (15.6% recommended for PEP), 30 autopsy personnel (50% recommended for PEP), and 21 other persons (4.8% recommended for PEP). Donor contacts represented 188 assessed with 20.2% recommended for PEP, compared with 5.6% of 306 recipient contacts recommended for PEP. Human rabies cases result in substantial use of public health and medical resources, especially when diagnosis is delayed. Although rare, clinicians should consider rabies in cases of encephalitis of unexplained aetiology, particularly for cases that may result in organ donation.     

Feed intake and serum GH, LH and cortisol in gilts after intracerebroventricular or intravenous injection of urocortin

In three experiments (Exp), ovariectomized gilts received intracerebroventricular (ICV; Exp 1 - with restraint, Exp 2 - without restraint) or intravenous (i.v.; Exp 3) injections of urocortin or saline to assess effects on feed intake and serum GH, LH, and cortisol. Following a 20-hr fast, feed was presented at 1 hr (Exp 1) or 30 min (Exp 2 and 3) after injection (time = 0 hr) of saline or 5 (U5) or 50 (U50) μg/pig (Exp 1 and 2) or 5 μg/kg BW (Exp 3) of urocortin. Blood samples were collected every 15 min from –2 to 6 hr relative to injection and hormone data pooled 2 hr before and hourly after treatment. Treatment with U50 decreased feed intake, relative to saline (treatment x time interaction; P < 0.05), when delivered ICV but not i.v. A treatment by time interaction was detected for GH (Exp 1, 2, 3) and LH (Exp 1 and 2; P < 0.01). Serum GH increased over time (relative to −2 hr; P < 0.05) following treatment with urocortin but not saline regardless of route of administration. Conversely, in Exp 1 (U5 and U50) and Exp 2 (U50), LH decreased relative to −2 hr with a delayed decrease during Exp 1. Serum cortisol was not affected by treatment in Exp 1, but increased following urocortin in Exp 2 and 3 (treatment by time interaction, P < 0.01). These data provide evidence that urocortin modulates GH and LH concentrations and suppresses feed intake in gilts via mechanisms which may be independent of cortisol and may depend upon dose and route of administration.     

Life cycle of Eimeria procera in experimentally infected grey partridges (Perdix perdix)

We examined exogenous and endogenous development of Eimeria procera in experimentally infected grey partridges (Perdix perdix). Our examination included data on morphology, localization, duration of schizogony and gametogony and morphology of sporulated oocysts. The endogenous stages of E. procera developed in large numbers within the epithelial cells of caecal crypts. The asexual development comprised three generations of schizonts. The first fully developed macrogametes and microgamonts were observed on Day 5 post-infection (p.i.) in histologic section. The patent period began on Day 6 p.i. and ended on Day 11 p.i. with peak production of oocysts on Days 7 and 8. Long oval oocysts of E. procera measured 25.78–28.13 μm in length and 14.06–15.24 μm in width, sporulation time ranged from 18 to 24 h at 25°C and from 36 to 48 h at 20°C.