While limbal epithelial cells are used for treating ocular surface wounds, the therapeutic potential of mesenchymal cells cultivated from the limbal stroma (LMSC) is less clear. We have therefore ...examined the effects of LMSC when applied to acute ocular surface wounds. LMSC derived from male rabbits (RLMSC) were applied to the ocular surface of female rabbits immediately following removal of the corneal and limbal epithelium. Human amniotic membrane (HAM) was used as the vehicle for implanting the RLMSC. The effects of RLMSC were examined when applied alone (n = 3) and in conjunction with a stratified culture of human limbal epithelial cells (HLE) grown on the opposing surface of the HAM (n = 3). Outcomes were monitored over 3 months in comparison with animals receiving no treatment (n = 3) or treatment with HLE alone on HAM (n = 3). Animals treated with RLMSC (n = 6) displayed faster re-epithelialization (∼90% versus 70% healing after 12 weeks), with best results being observed when RLMSC were pre-cultivated and implanted in the presence of HLE (p < 0.01; 90% healing by 7 weeks). While all animals displayed conjunctival cells on the corneal surface (by presence of goblet cells and/or keratin 13 expression) and corneal neovascularization, evidence of corneal epithelial regeneration was observed in animals that received RLMSC in the presence of HLE (by staining for keratin 3 and the absence of goblet cells). Conversely, corneal neovascularization was significantly greater when RLMSC were applied in the absence of HLE (<0.05; 90% of cornea compared with 20–30% in other cohorts). Nevertheless, neither human nuclear antigen nor rabbit Y chromosome were detected within the regenerated epithelium. Our results demonstrate that while cultured LMSC encourage corneal re-epithelialization, healing is improved by the pre-cultivation and implantation of these mesenchymal cells in the presence of limbal epithelial cells.
Comparisons were made of carcass and meat quality characteristics of pasture-raised Texel-cross ewe (
n
=
269) and ram (
n
=
275) lambs between 5 and 8 months of age with an average carcass weight of ...17.2
kg. Carcass assessment was based on linear dimensions and dissection of a leg into muscle, fat and bone, and meat quality measurements were made on
M. semimembranosus and
M. longissimus. At a set carcass weight, ewe lambs had higher dressing percentages (2%), shorter carcasses (0.7
cm), and heavier leg cuts (35
g) (
P
<
0.01) than males. At the same leg weight, legs of ewe lambs were fatter than males (subcutaneous plus intermuscular fat; 11.2% vs 9.6%;
P
<
0.001), whereas legs of ram lambs contained significantly more muscle and bone than females (
P
<
0.001). Leg muscle to bone ratio (4.7 vs 4.4) and muscularity were higher for females than males (
P
<
0.001). However, the relationship between leg muscle to bone ratio and muscularity was not the same between the sexes, and for any given muscularity value the muscle to bone ratio of ram lambs was lower than females. As a result, if carcass lean meat yield is predicted from a measure of carcass shape, such as muscularity, lean meat yields will be overestimated for males and underestimated for females. Meat quality was lower in ram lambs than in females (
P
<
0.001) as shown by higher Warner-Bratzler shear values (peak value 109.8 vs 97.0
N for
M. Semimembranosus), higher ultimate meat pH values, and lower redness (
a
∗) and lightness (
L
∗, for the longissimus muscle only) values (
P
<
0.001). It is concluded that significant differences between ewe and ram lambs do exist for many carcass and meat quality traits, but for most quality traits the differences are small.
Genetic and phenotypic parameters were estimated for production and disease traits (including dagginess) from about 2 million pedigree-recorded animals born between 1990 and 2008 in New Zealand ...dual-purpose ram breeding flocks. This is the most comprehensive study of genetic parameter estimates for the New Zealand sheep industry to date and includes estimates that have not previously been reported. Estimates of heritability were moderate for BW at 8 mo (LW8), fleece weight at 12 mo (FW12), dagginess score at 3 and 8 mo (DAG3, DAG8; 0.31 to 0.37), typical for weaning weight (WWT), fecal egg count in summer (FEC1) and autumn (FEC2), and analogous Nematodirus counts (NEM1, NEM2; 0.17 to 0.21), and low for number of lambs born to ewes (NLB; 0.09). The genetic correlations among production traits, WWT, LW8, and FW12, were positive and moderate to high. Correlations of DAG3 and DAG8 with production and disease traits were low and mostly negative. The NLB had low, but typically positive, correlations with other traits. Disease traits also had low, but positive, correlations with production traits (WWT, LW8, and FW12), and were highly correlated among themselves. In general, the heritability estimate for BW and dagginess were greater than what is currently used in the New Zealand genetic evaluation service (Sheep Improvement Limited), and the availability of accurate estimates for dagginess plus parasite resistance and their genetic correlations with production traits will enable more accurate breeding values to be estimated for New Zealand sheep.
Western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) is highly sensitive to orally delivered double‐stranded RNA (dsRNA). RNAi in WCR is systemic and spreads throughout the insect ...body. This raises the question whether transitive RNAi is a mechanism that functions in WCR to amplify the RNAi response via production of secondary siRNA. Secondary siRNA production is achieved through RNA‐dependent RNA polymerase (RdRP) activity in other eukaryotic organisms, but RdRP has not been identified in WCR and any other insects. This study visualized the spread of the RNAi‐mediated knockdown of Dv v‐ATPase C mRNA throughout the WCR gut and other tissues using high‐sensitivity branched DNA in situ hybridization. Furthermore, we did not detect either secondary siRNA production or transitive RNAi in WCR through siRNA sequence profile analysis. Nucleotide mismatched sequences introduced into either the sense or antisense strand of v‐ATPase C dsRNAs were maintained in siRNAs derived from WCR fed with the mismatched dsRNAs in a strand specific manner. The distribution of all siRNAs was restricted to within the original target sequence regions, which may indicate the lack of new dsRNA synthesis leading to production of secondary siRNA. Thus, the systemic spread of RNAi in WCR may be derived from the original dsRNA molecules taken up from the gut lumen. These results indicate that the initial dsRNA dose is important for a lethal systemic RNAi response in WCR and have implications in developing effective dsRNA traits to control WCR and in resistance management to prolong the durability of RNAi trait technology.
A directed search for QTL affecting carcass traits was carried out in the region of growth differentiation factor 8 (GDF8, also known as myostatin) on ovine chromosome 2 in seven Texel-sired half-sib ...families totaling 927 progeny. Weights were recorded at birth, weaning, ultrasound scanning, and slaughter. Ultrasonic measures of LM cross-sectional dimensions and s.c. fat above the LM were made, with the same measurements made on the LM after slaughter. Following slaughter, linear measurements of carcass length and width were made on all carcasses, and legs and loins from 540 lambs were dissected. Genotyping was carried out using eight microsatellite markers from FCB128 to RM356 on OAR 2 and analyzed using Haley-Knott regression. There was no evidence for QTL for growth rates or linear carcass traits. There was some evidence for QTL affecting LM dimensions segregating in some sire families, although it was not consistent between ultrasound and carcass measures of the same traits. There was strong and consistent evidence for a QTL affecting muscle and fat traits in the leg that mapped between markers BM81124 and BULGE20 for the four sires that were heterozygous in this region, but not for the three sires that were homozygous. The size of the effect varied across the four sires, ranging from 0.5 to 0.9 of an adjusted SD for weight-adjusted leg muscle traits, and ranging from 0.6 to 1.2 of an adjusted SD for weight-adjusted leg fat traits. The clearest effect shown was for multivariate analysis combining all leg muscle and fat traits analyzed across sires, where the -log₁₀ probability was 14. Animals carrying the favorable haplotype had 3.3% more muscle and 9.9% less fat in the leg relative to animals carrying other haplotypes. There was evidence for a second peak in the region of marker TEXAN2 for one sire group. It seems that a QTL affecting muscle and fat traits exists within the New Zealand Texel population, and it maps to the region of GDF8 on OAR2.
Genetic and phenotypic parameters were estimated for dagginess, breech, wool, and fiber traits from approximately 29,500 progeny born in 2009 and 2010 in New Zealand dual-purpose ram breeding sheep ...flocks. Dagginess is adherence of fecal matter to the wool, and this study investigates the genetic and phenotypic correlations between dagginess and breech and wool traits. Estimates for heritability were moderate (0.21 to 0.44) for the following traits: dag score at 3 and 8 mo (DAG3, DAG8), breech bareness, wool length, wool bulk (BULK), mean fiber diameter, mean fiber diameter SD, mean fiber diameter CV, curvature (CURV), weaning weight at 3 mo, and autumn BW. Heritability estimates for fleece weight at 12 mo and proportion of medullated fibers were high (0.49 and 0.53, respectively). Dag score at 3 mo and DAG8 had low genetic and phenotypic correlations with all traits. Breech bareness had positive genetic and phenotypic correlations with CURV and BULK and mostly negative genetic correlations with all other wool traits. In summary the quantity and attributes of wool were not primary causative factors in fecal accumulation, leaving fecal consistency and composition as the major factors.
AIM: To determine flystrike incidence, and estimate genetic parameters and potential indicator traits to reduce breech strike susceptibility in Romney sheep in New Zealand. METHODS: Seventeen ...Romney-based flocks, on 11 farms from throughout New Zealand, were enrolled in a case-control study in 2009/10 and 2010/11. Farmers observed lambs with flystrike and recorded dag score, breech bareness score and flystrike location for each lamb. Flystrike location was classed as breech, body, belly, shoulders, head or feet. Paternity of lambs was determined from tissue samples collected from lambs and all potential sires on farms. Control lambs without flystrike were selected by matching birth year, flock and sex. Due to the majority of strike occurring in the breech, genetic parameters for breech strike were explored. Dag score and breech bareness were investigated as indirect indicators of breech strike. Heritabilities and genetic and phenotypic correlations were estimated for breech strike, dag score and breech bareness using an animal model. RESULTS: For the 2009/10 season, 484 cases of flystrike were recorded with mean incidence rate per farm of 1.76 (min 0.47, max 2.95)%. For the 2010/11 season, 352 cases were recorded with a mean incidence rate per farm of 2.54 (min 0.43, max 8.18)%. Over both years 694/792 (88%) cases of flystrike occurred on the breech. Heritability on the observed scale for breech strike was 0.32 (SE 0.10). Heritabilities for dag score and breech bareness were 0.23 (SE 0.09) and 0.35 (SE 0.11), respectively. Breech strike had a high positive genetic correlation with dag score (0.71) and a low negative genetic correlation with breech bareness (−0.17). Breech strike had a high phenotypic correlation with dag score (0.62) and negative phenotypic correlation with breech bareness (−0.06). CONCLUSIONS: The high genetic and phenotypic correlations between breech strike and dag score makes dag score a viable option for indirect selection for breech strike resistance in Romney sheep in New Zealand. The heritability, genetic and phenotypic correlations require validation in other dual-purpose breeds, before breeding values for breech strike can be implemented, for use throughout the New Zealand sheep industry.
A QTL affecting leg muscle and fat traits has been identified within the New Zealand Texel population. The QTL maps to a region on OAR 2 with a two-marker haplotype test established at markers ...BULGE20 and BM81124. These markers encompass the likely position of Growth Differentiation Factor 8 (GDF8). The pleiotropic effects of this QTL on meat quality traits are tested. Objective measures of meat quality including pH, color (L*, a*, and b*), and tenderness (as assessed by Warner-Bratzler shear force measurements) were assessed on longissimus and semi-membranosus muscles of 540 progeny from six Texel sires. Four of these sires were subsequently identified as segregating for leg muscle and fat traits. For these segregating sires, comparison of progeny that had inherited the favorable haplotype from their sire with those that had received the alternate haplotype revealed no significant differences in the meat quality traits assessed. This finding suggests that the muscling QTL does not have pleiotropic effects on meat quality. A general scan for meat quality QTL was carried out using genotype data for eight markers from FCB128 to RM356 flanking 122cM of OAR 2 using Haley-Knott regression. This analysis revealed two QTL for a single sire. A QTL detected in the region of Marker INRA40 for color L* mapped to a site close to the muscling QTL, but there was evidence to suggest it is at a distinct locus. The QTL in the region of Marker RM356 might map distal to Marker RM356, as no peak was observed. This QTL, which seems to affect pH, color a*, color b*, and Warner-Bratzler shear measurements, requires further characterization.
Genetic and phenotypic parameters were estimated for dagginess, breech, wool, and fiber traits from approximately 29,500 progeny born in 2009 and 2010 in New Zealand dual-purpose ram breeding sheep ...flocks. Dagginess is adherence of fecal matter to the wool, and this study investigates the genetic and phenotypic correlations between dagginess and breech and wool traits. Estimates for heritability were moderate (0.21 to 0.44) for the following traits: dag score at 3 and 8 mo (DAG3, DAG8), breech bareness, wool length, wool bulk (BULK), mean fiber diameter, mean fiber diameter SD, mean fiber diameter CV, curvature (CURV), weaning weight at 3 mo, and autumn BW. Heritability estimates for fleece weight at 12 mo and proportion of medullated fibers were high (0.49 and 0.53, respectively). Dag score at 3 mo and DAG8 had low genetic and phenotypic correlations with all traits. Breech bareness had positive genetic and phenotypic correlations with CURV and BULK and mostly negative genetic correlations with all other wool traits. In summary the quantity and attributes of wool were not primary causative factors in fecal accumulation, leaving fecal consistency and composition as the major factors. PUBLICATION ABSTRACT
Genetic and phenotypic parameters were estimated for production and disease traits (including dagginess) from about 2 million pedigree-recorded animals born between 1990 and 2008 in New Zealand ...dual-purpose ram breeding flocks. This is the most comprehensive study of genetic parameter estimates for the New Zealand sheep industry to date and includes estimates that have not previously been reported. Estimates of heritability were moderate for BW at 8 mo (LW8), fleece weight at 12 mo (FW12), dagginess score at 3 and 8 mo (DAG3, DAGS; 0.31 to 037), typical for weaning weight (WWT), fecal egg count in summer (FEC1) and autumn (FEC2), and analogous Nematodirus counts (NEM1, NEM2; 0.17 to 0.21), and low for number of lambs born to ewes (NLB; 0.09). The genetic correlations among production traits, WWT, LW8, and FW12, were positive and moderate to high. Correlations of DAG3 and DAG8 with production and disease traits were low and mostly negative. The NLB had low, but typically positive, correlations with other traits. Disease traits also had low, but positive, correlations with production traits (WWT, LW8, and FW12), and were highly correlated among themselves. In general, the heritability estimate for BW and dagginess were greater than what is currently used in the New Zealand genetic evaluation service (Sheep Improvement Limited), and the availability of accurate estimates for dagginess plus parasite resistance and their genetic correlations with production traits will enable more accurate breeding values to be estimated for New Zealand sheep. PUBLICATION ABSTRACT
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