A B-cell targeting virus disrupts potentially protective genomic methylation patterns in lymphoid tissue by increasing global 5-hydroxymethylcytosine levels

The mechanisms by which viruses modulate the immune system include changes in host genomic methylation. 5-hydroxymethylcytosine (5hmC) is the catalytic product of the Tet (Ten-11 translocation) family of enzymes and may serve as an intermediate of DNA demethylation. Recent reports suggest that 5hmC may confer consequences on cellular events including the pathogenesis of disease; in order to explore this possibility further we investigated both 5-methylcytosine (5mC) and 5hmC levels in healthy and diseased chicken bursas of Fabricius. We discovered that embryonic B-cells have high 5mC content while 5hmC decreases during bursa development. We propose that a high 5mC level protects from the mutagenic activity of the B-cell antibody diversifying enzyme activation induced deaminase (AID). In support of this view, AID mRNA increases significantly within the developing bursa from embryonic to post hatch stages while mRNAs that encode Tet family members 1 and 2 reduce over the same period. Moreover, our data revealed that infectious bursal disease virus (IBDV) disrupts this genomic methylation pattern causing a global increase in 5hmC levels in a mechanism that may involve increased Tet 1 and 2 mRNAs. To our knowledge this is the first time that a viral infection has been observed to cause global increases in genomic 5hmC within infected host tissues, underlining a mechanism that may involve the induction of B-cell genomic instability and cell death to facilitate viral egress.     

Characteristics of free-roaming cats and their caretakers

OBJECTIVES: To describe the characteristics of unowned, free-roaming cats and their caretakers who participated in a trap-neuter-return (TNR) program and to determine the effect of the program on free-roaming cat colonies. DESIGN: Prospective study. SAMPLE POPULATION: 101 caretakers of 920 unowned, free-roaming cats in 132 colonies in north central Florida. RESULTS: Most (85/101; 84%) caretakers were female. The median age was 45 years (range, 19 to 74 years). Most (89/101; 88%) caretakers owned pets and of those, most (67/101; 66%) owned cats. The major reasons for feeding free-roaming cats were sympathy and love of animals. Most caretakers reported that the cats they cared for were too wild to be adopted, but many also reported that they considered the cats to be like pets. The total surveyed cat population was 920 before participation in TNR and 678 after TNR. Mean colony size was 7 cats before TNR and 5.1 cats after TNR. Most cats lived on the caretaker's property. At the time of the survey, 70% (644/920) of the cats had been neutered. CONCLUSIONS AND CLINICAL RELEVANCE: The decrease in the surveyed free-roaming cat population was attributed to a reduction in births of new kittens, adoptions, deaths, and disappearances. Recognition of the human-animal bond that exists between caretakers and the feral cats they feed may facilitate the development of effective control programs for feral cat populations.     

Transdermal absorption of a liposome-encapsulated formulation of lidocaine following topical administration in cats

OBJECTIVE: To determine plasma disposition after dermal application of a liposome-encapsulated formulation of lidocaine in cats. ANIMALS: 6 healthy adult cats with a mean (+/- SD) body weight of 4.1 +/- 0.44 kg. PROCEDURE: CBC determination and biochemical analysis of blood samples were performed for all cats. Cats were anesthetized by use of isoflurane, and catheters were placed IV in a central vein. The next day, blood samples were obtained from the catheters before and 1, 2, 3, 4, 6, 8, 10, 12, and 24 hours after applying a 4% liposome-encapsulated lidocaine cream (15 mg/kg) to a clipped area over the cephalic vein. Plasma concentrations of lidocaine were analyzed with a high-performance liquid chromatography assay. Results-Two cats had minimal transdermal absorption of lidocaine, with lidocaine concentrations below the sensitivity of the assay at all but 1 or 2 time points. In the other 4 cats, the median maximum plasma concentration was 149.5 ng/ml, the median time to maximum plasma concentration was 2 hours, and the median area under the concentration versus time curve from zero to infinity was 1014.5 ng.h/ml. CONCLUSIONS AND CLINICAL RELEVANCE: Maximum plasma concentrations of lidocaine remained substantially below toxic plasma concentrations for cats. On the basis of these data, topical administration of a liposome-encapsulated lidocaine formulation at a dose of 15 mg/kg appears to be safe for use in healthy adult cats.     

Proteome-determined type-specific proteins of Mycobacterium avium subspecies paratuberculosis

Mycobacterium avium subspecies paratuberculosis (M. a. paratuberculosis) is a pathogen of ruminants, causing paratuberculosis (characterized by severe emaciation). The disease is endemic in many countries including the UK and places a severe economic burden on the global livestock industry. Two types of M. a. paratuberculosis can be classified by pulsed-field electrophoresis (I/III and II), which are phenotypically distinct and appear to have different host preferences. Proteomes of Type I and Type II M. a. paratuberculosis were analyzed by 2-D gel electrophoresis to determine if any significant differences existed between the subtypes. Seven different strains of Type I and 18 strains of Type II were analyzed and compared to detect type-specific differences. These 'type-specific' differences existed regardless of growth phase and were also exhibited in cells isolated directly from pathogenic lesions. Twenty-three spots predominated on the Type I profile, from which 17 proteins were identified. Twenty-one spots predominated on the Type II profile, from which 16 proteins were identified. None of the proteins identified as differentially represented on the profiles of Type I or Type II corresponded to open reading frames of the defining genomic regions as previously described for the Type I (sheep) and Type II (cattle). Sequence polymorphisms existing in Type I and II strains were identified in some open reading frames or regulatory regions of genes that correspond to proteins expressed in a type-specific fashion. The consequence of these is discussed in relation to protein expression and their impact on the type phenotype is discussed.     

Effects of intraruminal administration of zinc on gastric acid secretion in sheep.

Intraruminal administration of zinc sulphate at 100 and 200 mg Zn/kg bodyweight resulted in central and peripheral effects in sheep. Feed intake was reduced, pH of the duodenal contents elevated and the secretion of acid from isolated pouches of the abomasum doubled. Suggested explanations include a local inhibitory effect of zinc on abomasal acid secretion elevating duodenal pH and a resultant increased release of gastrin stimulating secretion from the pouches.