Biodistribution and Clearance of Intra-articular Liposomes in a Large Animal Model Using a Radiographic Marker

The intra-articular (IA) route of administration in treating arthritis has potential for targeting drug delivery to affected tissues, thereby minimising the attendant side-effects of systemically administered drugs. The ultra-structure of the synovium however facilitates rapid drug efflux from the joint; effectively the IA route is equivalent to other non-IV parenteral routes with regards absorption and redistribution into the systemic circulation. The aim of this study was to extend the drug residence time within the knee joint by using a liposome formulation. DPPC-based liposomes were prepared with the radio contrast agent iohexol as a drug marker. 8 sheep had their right knees injected IA with iohexol liposomes and the contralateral joints with either free iohexol or empty liposomes. Joints were radiographed at multiple time points up to 16 days post-injection. Iohexol-mediated radiopacity was quantified by densitometer. Sheep were sacrificed at the end of the study for microscopy of synovial tissues.

Good visualization of iohexol-mediated radiopacity with fine anatomical definition was possible throughout the experiment. Also evident on the films was extra-articular radiopacity with liposomes tracking along muscle facial planes. Cellular and tissue localization with light microscopy was possible through use of frozen sections and because of the large liposome size.

Residence of encapsulated iohexol within the knee joint was greatly prolonged.

Liposomal iohexol declined bi-exponentially with a terminal elimination half-life of 134 hours. In contrast, free iohexol was undetectable @ 3 hours post-injection.     

Non-genetic sources of variation of milk production and reproduction and interactions between both classes of traits in Sicilo-Sarde dairy sheep

This work aimed to study the sources of variation in productive and reproductive traits of the dairy Sicilo-Sarde ewes and to further investigate the interaction between both classes of traits. After edits, a database containing 5935 lactation records collected during 6 successive years in eight dairy flocks in the North of Tunisia was used. Total milked milk (TMM) in the milking-only period was retained as productive trait. The interval from the start of the mating period to the subsequent lambing (IML) and the lambing status (LS) were designed as reproductive traits. Sicilo-Sarde ewes had an average TMM of 60.93 l (±44.12) during 132.8 days (±46.6) after a suckling period of 100.4 days (±24.9). Average IML was 165.7 days. In a first step, the major factors influencing milk production and reproductive traits were determined. The significant sources of variation identified for TMM were: flock, month of lambing, year of lambing, parity, suckling length, litter size and milking-only length. Flock×month of the start of the mating period, parity, year of mating and litter size were identified as significant factors of variation for IML, while flock×month of the start of the mating period, parity and year of mating were identified as significant sources of variation for LS. In a second step, variance components were estimated using a three traits threshold mixed model, which combined LS as categorical trait and TMM and IML as continuous traits. Repeatability estimates were 0.21 (±0.03) for TMM, 0.09 (±0.02) for IML, and 0.10 (±0.05) for LS. Moreover, TMM and IML were found to be favorably associated for the flock× year of lambing effect (−0.45±0.18) but unfavorably associated for the animal effect (0.20±0.09).     

An improved method for typing of plasma α1-protease inhibitor (PI1) variants in sheep

Summary
Sheep plasma α₁-protease inhibitor (PI1) variants were typed by analysing neuraminidase treated samples by isoelectric focusing in ultra-thin polyacrylamide gels followed by staining for trypsin inhibition. The 10 different PI1 pheno-types observed were shown to be controlled by four codominant alleles. The probability of sire exclusion provided by PI1 polymorphism was about 0.45 in both of the studied milk sheep breeds (Latxa and Karrantzar) of the Basque Country.     

Identification of Prolificacy‐Related Differentially Expressed Proteins from Sheep (Ovis aries) Hypothalamus by Comparative Proteomics

Reproduction, as a physiologically complex process, can significantly affect the development of the sheep industry. However, a lack of overall understanding to sheep fecundity has long blocked the progress in sheep breeding and husbandry. In the present study, the aim is to identify differentially expressed proteins (DEPs) from hypothalamus in sheep without FecB mutation in two comparison groups: polytocous (PF) versus monotocous (MF) sheep at follicular phase and polytocous (PL) versus monotocous (ML) sheep at luteal phase. Totally 5058 proteins are identified in sheep hypothalamus, where 22 in PF versus MF, and 39 proteins in PL versus ML are differentially expressed, respectively. A functional analysis is then conducted including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis to reveal the potential roles of these DEPs. The proteins ENSOARP00000020097, ENSOARP00000006714, growth hormone (GH), histone deacetylase 4 (HDAC4), and 5′‐3′ exoribonuclease 2 (XRN2) in PF versus MF, and bcl‐2‐associated athanogene 4 (BAG4), insulin‐like growth factor‐1 receptor (IGF1R), hydroxysteroid 11‐beta dehydrogenase 1 (HSD11B1), and transthyretin (TTR) in PL versus ML appear to modulate reproduction, presumably by influencing the activities of gonadotropin‐releasing hormone (GnRH). This study provides an alternative method to identify DEPs associated with sheep prolificacy from the hypothalamus. The mass spectrometry data are available via ProteomeXchange with identifier PXD013822.     

藏山羊KLF8基因克隆及组织表达分析

本试验旨在获得藏山羊KLF8基因序列,并分析其生物学特征,同时阐明该基因在不同组织中的表达情况。利用RT-PCR技术克隆藏山羊KLF8基因序列,利用实时荧光定量PCR (quantitative real-time PCR,qPCR)检测其在藏山羊各个组织中的表达丰度。结果表明,获得藏山羊KLF8基因序列1 069 bp,其中包含CDS区1 008 bp,5'UTR序列28 bp和3'UTR序列33 bp,共编码335个氨基酸,为不稳定亲水碱性蛋白。KLF8基因在藏山羊的肺脏组织中表达水平最高,极显著高于其他组织(p<0.01)。本研究为进一步阐明KLF8基因在藏山羊中的生物学功能提供了依据。     

Net mineral requirements for the growth and maintenance of Somali lambs

Minerals are limiting factors in animal production, and the knowledge of mineral requirements for livestock is crucial to the success of a commercial enterprise. Hair sheep may have different mineral requirements than those presents by the international committees. A study was carried to evaluate the net calcium (Ca), phosphorus (P), magnesium (Mg), sodium (Na), potassium (K), zinc (Zn), iron (Fe), manganese (Mn) and copper (Cu) requirements for the growth and maintenance of Brazilian Somali lambs. A total of 48 hair lambs (13.5±1.8 kg) aged 60±15 days were allocated to individual pens. Eight animals were slaughtered at the beginning of the experiment to serve as a reference group to estimate initial empty BW (EBW) and initial body composition. The remaining lambs (n=40) were assigned to a completely randomized design with eight replications in five levels of metabolizable energy (ME; 4.93, 8.65, 9.41, 10.12 and 11.24 MJ/kg DM). When the lambs of a given treatment reached an average BW of 28 kg, they were slaughtered. Initial body composition was used to calculate the retention of minerals. Mineral body composition was fit using a logarithmic equation in the form of a nonlinear model. The maintenance requirements were estimated from regressions of mineral retention in the empty body on mineral intake. The body mineral concentration decreased in lambs with a BW ranging from 15 to 30 kg. The net mineral requirements (100 g/day of average daily gain (ADG)) decreased from 0.52 to 0.51 g for Ca, 0.28 to 0.23 g for P, 0.02 to 0.02 g for Mg, 0.09 to 0.08 g for Na, 0.11 to 0.09 g for K, 1.30 to 1.08 mg for Zn, 3.77 to 3.22 mg for Fe, 0.08 to 0.06 mg for Mn and 0.09 to 0.08 mg for Cu when BW increased from 15 to 30 kg. The daily net requirements for maintenance per kilogram of BW were 30.13 mg of Ca, 27.58 mg of P, 1.26 mg of Mg, 4.12 mg of Na, 8.11 mg of K, 0.133 mg of Zn, 0.271 mg of Fe, 0.002 mg of Mn and 0.014 mg of Cu. The results of this study indicate that the net mineral requirements for weight gain and maintenance in Brazilian Somali lambs are different than the values that are commonly recommended by the main evaluation systems for feed and nutritional requirements for sheep. These results for the nutritional requirements of minerals may help to optimize mineral supply for hair sheep.     

Water footprint assessment of sheep farming systems based on farm survey data

Water scarcity is among the main challenges making vulnerable the livestock farming systems in drylands. The water footprint (WF) indicator was proposed as a metric to measure the impacts of livestock production on freshwater resources. Therefore, this study aimed to assess water use in five different Tunisian sheep production systems using the Water Footprint Network methodology. The primary data were obtained from 1050 sheep farms located in 13 Tunisian provinces. A multivariate analysis was performed to characterize the different farming systems. A validation step of the WF modeled values of sheep meat was conducted in 12 sheep farms belonging to two different farming systems. This was done through year-round monitoring of on-farm practices using water metres and recording equipment’s taking into account the direct and indirect water use. The typology analysis came up with five sheep farming systems that are the mixed sheep-cereal (MSC), the agro-sylvo-pastoral (ASP), the agro-pastoral (AP), the extensive agro-pastoral (EAP) and the mixed sheep-olive tree farming systems. The WF of sheep meat produced under the target farming systems ranged from 8654 to 13 056 l/kg live weight. The evaluation of WF of five different sheep production systems figured out that sheep raised under the EAP farming system had the greatest WF per ton of live animal. However, the ASP farming system exhibited the lowest WF. Water used to grow feedstuffs for sheep production accounts for 98% of the total WF of sheep. The green WF accounts for more than 92% of the total WF in all farming systems. Results of monitoring water use at farm scale show that the modeled values of WF are overestimated by an average of 23.3% and 24.1% for the selected farms assigned to the MSC and AP farming systems, respectively. Water use for sheep production is high in most of the Tunisian farms. Therefore, the general assumption that ‘meat production is a driver of water scarcity’ is supported and should be considered as an important focal point in agricultural and water policies. Particular attention should be given to forage crops with low WFs and high contribution to dry matter to provide ration with low WF. The efficient use of green water along the meat value chain is essential to minimize the depletion of blue water resources and to reduce the economic dependency on virtual water through the import of feedstuffs.     

Meta-analysis of spineless cactus feeding to meat lambs: performance and development of mathematical models to predict dry matter intake and average daily gain

Spineless cactus is a useful feed for various animal species in arid and semiarid regions due to its adaptability to dry and harsh soil, high efficiency of water use and carbohydrates storage. This meta-analysis was carried out to assess the effect of spineless cactus on animal performance, and develop and evaluate equations to predict dry matter intake (DMI) and average daily gain (ADG) in meat lambs. Equations for predicting DMI and ADG as a function of animal and diet characteristics were developed using data from eight experiments. The dataset was comprised of 40 treatment means from 289 meat lambs, in which cactus was included from 0 to 75% of the diet dry matter (DM). Accuracy and precision were evaluated by cross-validation using the mean square error of prediction (MSEP), which was decomposed into mean bias, systematic bias and random error; concordance correlation coefficient, which was decomposed into accuracy (C_b) and precision (ρ); and coefficient of determination (R²). In addition, the data set was used to evaluate the predicting accuracy and precision of the main lamb feeding systems (Agricultural and Food Research Council, Small Ruminant Nutritional System, National Research Council and Institut National de la Recherche Agronomique) and also two Brazilian studies. The DMI, CP intake (CPI), metabolizable energy (ME) intake and ADG increased when cactus was included up to 499 g/kg DM (P<0.001). In contrast, animals fed high levels of cactus (>500 g/kg DM) had a decreased DMI, CPI and NDF intake, but increased feed efficiency (P<0.001) and similar ADG compared with those without cactus addition. The DMI was positively correlated with initial BW, final BW, concentrate and ADG, while it was negatively correlated with cactus inclusion and ME of the diet. On other hand, ADG was positively correlated with DMI, initial and mean BW and concentrate, and it was negatively correlated with cactus inclusion. The two developed equations had high accuracy (C_b of 0.95 for DMI and 0.94 for ADG) and the random error of MSEP was 99% for both equations. The precision of both equations was moderate, with R² values of 0.53 and 0.50 and ρ values of 0.73 and 0.71 for DMI and ADG, respectively. In conclusion, the developed equation to predict DMI had moderate precision and high accuracy, nonetheless, it was more efficient than those reported in the literature. The proposed equations can be a useful alternative to estimate intake and performance of lambs fed cactus.