Sequence of the domestic duck (Anas platyrhynchos) growth hormone-encoding gene and genetic variation in the promoter region

The duck growth hormone encoding gene and its promoter region were amplified by polymerase chain reaction (PCR). A total of 5.25 kb were cloned and sequenced. Duck growth hormone (GH) consists of five exons and four introns and is structurally similar to mammalian and chicken GH gene. Although the distal region of duck GH promoter showed no similarity to chicken and turkey promoters, the proximal region of the promoter contained two putative Pit‐1 binding sequences, and showed similarity to chicken and turkey GH promoters. Genetic variation was detected at five positions of the promoter region. The results of this study indicate that the expression of duck GH is likely regulated in a similar manner to that of chicken GH via enhancer‐type cis‐acting elements and the presence of genetic variation in the duck GH gene may be applicable to marker‐assisted selection.     

The effect of the Naked Neck genotype (<Emphasis Type="Italic">Nana</Emphasis>), feeding and outdoor rearing on growth and carcass characteristics of free range broilers in a hot climate

Alternative poultry production with special reference to free range broilers has increased significantly since the nineties in many regions of the world. Numerous factors influence the productive performance of this type of broilers: genotype (namely the use of naked neck animals), feeding and access to an outdoor area. The aim of this paper is to study the influence of each of these factors on the productive performance of free range broilers under commercial rearing conditions. A total of 3200, day old chicks of both sexes from naked neck and normally feathered genotypes were used in this trial. After a joint initiation phase, animals were divided into four different treatments with the combination of two concentrates (high vs low energy content) and management (access to outside park or not). Experiment lasted a total of 12 weeks. Live weight date was recorded weekly and a samples of animals from the trial were sacrificed at the age of 8, 10 and 12 weeks, when carcass characteristics were determined. Besides sex, the only factor that seems to affect growth characteristics was genotype as naked neck animals had poorer growth rates than normally feathered. No effect was detected on carcass yields and percentages of carcass components for any of the variables. From the data presented in this trial the practises associated with free range production are of relative inconsequence to the technical animal production parameters and can only be justified by a pressing need to differentiate these products from standard poultry products in what concerns both welfare issues and meat characteristics. The results also indicate that genetic material from alternative poultry production in Europe can be a useful option in poultry production development projects in the tropics.     

Scrotal circumference,bodyweight and semen characteristics in growing dairy-breed natural-service bulls in Tasmania,Australia

Aims: To provide herd managers with a set of decision rules allowing them to predict the likelihood that a juvenile bull is ready for Bull Breeding Soundness Evaluation (BBSE), or breeding, if bodyweight and scrotal circumference are known.

Methods: This was a longitudinal study following two groups of young pasture-fed Holstein and Jersey bulls from northwest Tasmania, Australia. Individual scrotal circumference, bodyweight and semen characteristics were recorded at 6–8 weekly intervals, from 6–18 months of age. Classification and regression tree analyses were used to predict the probability that a bull had ≥70% normal sperm morphology based on scrotal circumference and bodyweight measurements.

Results: Overall 1,661 scrotal circumference and bodyweight measurements were obtained, and 518 semen samples from 356 bulls were assessed for sperm morphology, from 16 examination sessions that took place between 29 May 2015 and 17 August 2016. Classification and regression tree analyses generated a decision tree for Holstein bulls with four node endpoints, and for Jersey bulls with three node endpoints. Diagnostic test performance showed that for Holstein bulls, using the node endpoints of scrotal circumference ≥27?cm and bodyweight ≥349?kg, 98% had ≥70% normal sperm (positive likelihood ratio 10.4; 95% CI?=?2.7–41), and using the node endpoints of scrotal circumference ≥27?cm and bodyweight between 282–349?kg, 89% had ≥70% normal sperm (positive likelihood ratio 1.6; 95% CI?=?0.9–2.6). For Jersey bulls, using the node endpoints of bodyweight ≥259?kg and scrotal circumference ≥29?cm, 88% had ≥70% normal sperm (positive likelihood ratio 3.4; 95% CI?=?1.6–7.0).

Conclusions: This study provides a set of relatively simple decision rules based on bodyweight and scrotal circumference measurements that allows herd managers to assess the likelihood that juvenile bulls are ready for BBSE or breeding.

Abbreviations: BBSE: Bull breeding soundness evaluation; BRT: Boosted regression tree     

Long‐term Treatment with Methylene Blue in a Dog with Hereditary Methemoglobinemia Caused by Cytochrome b5 Reductase Deficiency

A juvenile male mixed breed dog was presented for lethargy, exercise intolerance, and aggression when touched on the head. Cyanosis, tachycardia, and tachypnea were observed and persisted during oxygen supplementation. Arterial blood gas analysis by co‐oximetry identified an increased methemoglobin concentration (27%; normal, <2%) with normal arterial oxygen tension. The methemoglobinemia and associated clinical signs resolved after administration of methylene blue (1 mg/kg) IV, and the dog was discharged. The affected dog's whole‐genome sequence contained 2 potentially causal heterozygous CYB5R3 missense mutations suggesting that cytochrome b5 reductase deficiency was responsible for the methemoglobinemia. This hypothesis was confirmed by enzyme analysis that identified cytochrome b5 reductase activity in the affected dog's erythrocytes to only approximately 6% of that in a control sample. Clinical signs recurred 11 days after discharge but normalized and the methemoglobin concentration decreased with methylene blue administration PO (1.5 mg/kg, initially daily and then every other day).     

An Aqueous Extract of the Dry Mycelium of Penicillium chrysogenum Induces Resistance in Several Crops under Controlled and Field Conditions

We have examined the effect of Pen, an aqueous extract of the dry mycelium of Penicillium chrysogenum, on plant–pathogen interactions. Pen controlled a broad range of pathogens on several crop plants under greenhouse and field conditions. Pen protected grapevine from downy and powdery mildew (caused by Plasmopara viticola and Uncinula necator), tomato from early blight (caused by Phytophthora infestans), onion from downy mildew (Peronospora destructor) and apple trees from apple scab (caused by Venturia inaequalis) to a similar extent as fungicides such as copper and sulphur or well-known inducers such as benzothiadiazole or β-aminobutyric acid. Pen had no major direct fungicidal effect and is thus supposed to protect plants by activating their defense mechanisms. The raw material for extraction of Pen was available in constant quality, a prerequisite for commercial application. Under certain conditions, Pen caused phytotoxic side effects. The symptoms mostly consisted of small necrotic spots or, more rarely, of larger necrotic areas. The development of the symptoms was dependent on several parameters, including concentration of Pen, the number of applications, the persistence on the plant tissue, the plant species and variety and environmental conditions. In grapevine, a partially purified fraction of Pen was much less toxic than the crude Pen extract, but protected the plants to a similar extent against P. viticola. Our data show that Pen has interesting and unique properties as a plant protection agent, but more research is needed to further reduce its phytotoxic side effects.     

The Control of Reactive Oxygen Species Influences Porcine Oocyte In Vitro Maturation

The aim of this study was to examine the effect of varying intracellular reactive oxygen species (ROS) levels during oocyte in vitro maturation with enzymatic ROS production systems (xanthine + xanthine oxidase or xanthine + xanthine oxidase + catalase), scavenger systems (catalase or superoxide dismutase + catalase) or cysteine on porcine oocyte maturation. Oocyte ROS levels showed an increase when H₂O₂ or O₂?^‐ production systems were added to the culture medium (p < 0.05). On the other hand, the presence of ROS scavengers in the maturation medium did not modify oocyte ROS levels compared with the control after 48 h of maturation, but the addition of cysteine induced a decrease in oocyte ROS levels (p < 0.05). The ROS production systems used in this work did not modified the percentage of oocyte nuclear maturation, but increased the decondensation of sperm head (p < 0.05) and decreased the pronuclear formation (p < 0.05). In turn, the addition of O₂?^‐ and H₂O₂ scavenging systems during in vitro maturation did not modify the percentage of oocytes reaching metaphase II nor the oocytes with decondensed sperm head or pronuclei after fertilization. However, both parameters increased in the presence of cysteine (p < 0.05). The exogenous generation of O₂?^‐ and H₂O₂ during oocyte in vitro maturation would not affect nuclear maturation or later sperm penetration, but most of the spermatozoa cannot progress to form the pronuclei after fusion with the oocyte. The decrease in endogenous ROS levels by the addition of cysteine would improve pronuclear formation after sperm penetration.     

Immunohistochemical Localization of RANK, RANKL and OPG in Healthy and Arthritic Canine Elbow Joints

Objective— To determine if the receptor activator of nuclear factor-κB–receptor activator of nuclear factor-κB ligand–osteoprotegerin (RANK–RANKL–OPG) system is active in bone remodeling in dogs and, if so, whether differences in expression of these mediators occur in healthy and arthritic joints.
Study Design— Experimental study.
Sample Population— Fragmented processus coronoidei (n=20) were surgically removed from dogs with elbow arthritis and 5 corresponding healthy samples from dogs euthanatized for reasons other than elbow joint disease.
Methods— Bright-field immunohistochemistry and high-resolution fluorescence microscopy were used to investigate the distribution of RANK, RANKL, and OPG in healthy and arthritic joints.
Results— All 3 molecules were identified by immunostaining of canine bone tissue. In elbow dysplasia, the number of RANK-positive osteoclasts was increased. In their vicinity, cells expressing RANKL, a mediator of osteoclast activation, were abundant whereas the number of osteoblasts having the potential to limit osteoclastogenesis and bone resorption via OPG was few.
Conclusions— The RANK–RANKL–OPG system is active in bone remodeling in dogs. In elbow dysplasia, a surplus of molecules promoting osteoclastogenesis was evident and is indicative of an imbalance between the mediators regulating bone resorption and bone formation. Both OPG and neutralizing antibodies against RANKL have the potential to counterbalance bone resorption.
Clinical Relevance— Therapeutic use of neutralizing antibodies against RANKL to inhibit osteoclast activation warrants further investigation.     

Phosphofructokinase and Malate Dehydrogenase Participate in the In Vitro Maturation of Porcine Oocytes

Oocyte maturation depends on the metabolic activity of cumulus–oocyte complex (COC) that performs nutritive and regulatory functions during this process. In this work, the enzymes [phosphofructokinase (PFK) and malate dehydrogenase (MDH)] were tested to elucidate the metabolic profile of porcine COCs during the in vitro maturation (IVM). Enzymatic activity was expressed in U/COC and U/mg protein (specific activity) as mean ± SEM. In vitro maturation was performed with 2‐oxoglutarate (5, 10 and 20 mm ) or hydroxymalonate (30, 60 and 100 mm ) inhibitors of PFK and MDH, respectively. The PFK and MDH activities (U) remained constant during maturation. For PFK, the U were (2.48 ± 0.23) 10^?5 and (2.54 ± 0.32) 10^?5, and for MDH, the U were (4.72 ± 0.42) 10^?5 and (4.38 ± 0.25) 10^?5 for immature and in vitro matured COCs, respectively. The specific activities were significantly lower after IVM, for PFK (4.29 ± 0.48) 10^?3 and (0.94 ± 0.12) 10^?3, and for MDH (9.08 ± 0.93) 10^?3 and (1.89 ± 0.10) 10^?3 for immature and in vitro matured COCs, respectively. In vitro maturation percentages and enzymatic activity diminished with 20 mm 2‐oxoglutarate or 60 mm hydroxymalonate (p < 0.05). Viability was not affected by any concentration of the inhibitors evaluated. The U remained unchanged during IVM; however, the increase in the total protein content per COC provoked a decrease in the specific activity of both enzymes. Phosphofructokinase and MDH necessary for oocyte IVM would be already present in the immature oocyte. The presence of inhibitors of these enzymes impairs the meiotic maturation. Therefore, the participation of these enzymes in the energy metabolism of the porcine oocyte during IVM is confirmed in this study.