Molecular typing of Iranian field isolates Mycoplasma synoviae and their differentiation from the live commercial vaccine strain MS-H using vlhA gene

1. The single-copy domain of the N-terminal region of the vlhA gene of Mycoplasma synoviae was sequenced, analysed and verified and used to type Iranian field isolates of M. synoviae and the MS-H live vaccine strain. In addition, a restriction fragment length polymorphism (RFLP) method was developed to differentiate between field isolates of Iranian and MS-H vaccine strains.

2. All sequences were analysed and aligned; the percentage similarity of the DNA was calculated and dendrograms were constructed. Based on single nucleotide polymorphism (SNP) that existed in all field isolates in Iran, the PCR-RFLP method allowed the differentiation of all M. synoviae field isolates from the vaccine strain.

3. Using phylogenetic analysis, the isolates were assigned to 8 unique genotypes and, within each group, DNA had a high level of similarity.

4. DNA sequence analysis and PCR-RFLP of the amplicon based on percent similarity and evolutionary relationship appeared to be useful tools for strain differentiation whether M. synoviae clinical isolates from infected chickens were derived from the vaccine strain or wild-type strains.

5. This study confirms the potential value of strain typing for epidemiological purposes and suggests that phylogenetic studies are essential to understand the true relationships between strains.     

Comparing Genotypes of Different Body Sizes for Growth-related Traits in Chickens. II. Feed Consumption Performance under Choice Feeding Using Commercial Starter Ration and Whole-grain Wheat

Three genotypes of chickens including one commercial broiler (CB) and two others from the Norwegian gene bank [Barred Plymouth Rock (BPR) and Jærhøn (JH)] were raised from 1 day to 20 weeks of age with a free choice of a commercial starter ration and whole-grain wheat. Body weights and pen-feed intakes were recorded every 2 weeks. The weight–age data were fitted to a Gompertz growth equation where, among other parameters, adult body weights were estimated and used for size scaling. The heavier CB chickens consumed more food (starter and wheat) than the other two genotypes. The maximum proportions of whole-grain wheat in the diet were 0.56, 0.68 and 0.75 for BPR, CB and JH birds, respectively. Although the proportions of whole-grain wheat were similar between the genotypes as age progressed, their maintenance requirement and feed intakes relative to body weight suggested a major genetic role and a possible interplay of both genetic potential and body size.     

Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

Early heat acclimation during incubation improves Japanese quail performance under summer conditions

Effects of exposing quail eggs to high temperature on the heat tolerance ability and productivity of birds were investigated. Four groups of 600 fertile eggs were randomly selected; the first group was incubated under 37.5 °C and the hatched chicks were reared under a gradual decrease in temperature from 35 to 24 °C (Control). The second group was exposed to 39.1 °C for 2 h/day during 4–14 days of embryogenesis and the hatched chicks were reared under a gradual decrease in temperature from 35 to 24 °C. The third group was incubated under 37.5 °C and the hatched chicks were exposed to 39?±?1 °C for 2 h/day during 4–14 days of age. The fourth group was exposed to 39?±?1 °C for 2 h during 4–14 days of embryogenesis and the hatched chicks were exposed to 39?±?1 °C for 2 h/day during 4–14 days of age. The temperature applied changed (P?<?0.01) embryo weight and incubation period. Birds exposed to high temperature during brooding had superior growth performance, dressed carcass, body temperature and health traits. Birds subjected to 39?±?1 °C during brooding exhibited decreased feed consumption and body weight gain. Finally, this work suggests that thermal acclimation during embryogenesis might offer a practical method for easing heat stress.

     

Plasticity and stability in the major fatty acid content of almond kernels grown under two Mediterranean climates

Summary

Oil contents and fatty acid compositions were determined in 17 almond cultivars growing under two different Mediterranean conditions, in northeast Spain and in central Morocco. The major differences were irrigation management in Spain and higher temperatures in Morocco. Although a cultivar effect was observed for some compounds, the general trend was for an increase in total oil content [58.65% vs. 55.58% (w/w)] and in the percentage of oleic acid [71.1% vs. 68.62% (w/w)] in those kernels harvested in Spain, probably due to having lower temperatures and better water status. As the total oil content and level of oleic acid in almond oil are the major parameters for higher quality almond kernels, these results emphasise the need to optimise the management of almond orchards to improve kernel quality, not only from the commercial and industrial points of view, but also because of their healthgiving properties.     

A high virulence and pathotype diversity of Puccinia striiformis f.sp. tritici at its centre of diversity,the Himalayan region of Pakistan

Information on the pathogen virulence profile and diversity across locations is crucial for host germplasm improvement and deployment. The rapid acquisition of virulence to host resistance by the wheat yellow/stripe rust pathogen (Puccinia striiformis f.sp. tritici: PST), makes it crucial to know about its virulence and pathotype diversity. Recent studies have shown the plausible centre of origin of the pathogen in the Himalayan region, with Pakistan being the most ancestral to all other worldwide populations. To assess the status of virulence and pathotype diversity in the Himalayan region of Pakistan, a set of 127 PST infected wheat samples from eight locations were collected, multiplied and pathotyped using a set of 36 differential lines from the world set, European and Chinese sets, and 9 Avocet Yr isolines. Virulence (Vr) was recorded to 18 out of 24 tested yellow rust resistance (Yr) genes, while a total of 53 pathotypes were detected out of 127 isolates tested. Virulence was found to the resistance genes rarely deployed in Pakistan (Vr8) or even worldwide level (Vr5), while virulence to Vilmorin 23 (Yr3+) was absent in Pakistan, which is common in Europe. None of the pathotypes was dominant across all locations, however, no clear spatial structuring was observed for the studied locations. Our results suggested a high virulence and pathotype diversity in line with the previously proposed potential role of sexual recombination in the temporal maintenance of PST in the Himalayan region of Pakistan. This information should be useful in host resistance gene improvement and deployment.     

Cryopreservation of Persian sturgeon (Acipenser persicus) sperm: effects of cryoprotectants,antioxidant, membrane stabilizer,equilibration time and dilution ratio on sperm motility and fertility

Three experiments were performed to develop protocols for cryopreservation of Persian sturgeon Acipenser persicus, sperm. In the first experiment, sperm from six males was individually split in three subsamples and cryopreserved using Modified Tsvetkova's extender (mT) supplemented with dimethyl sulfoxide (DMSO), methanol (MeOH), glycerol (Gly) and ethylene glycol (EG) at concentration of 5%, 10%, 15% and 20%. In the second set of experiments, the effects of six equilibration times (0, 5, 10, 20, 40 and 60 min) and dilution ratios (volume sperm: volume extender 1:0.5, 1:1, 1:2, 1:3, 1:5 and 1:10) and the additive advantage of bovine serum albumin (BSA; 0, 2.5, 5 and 10 mg mL^?1) and ascorbic acid (0, 2.5, 5 and 10 U mL^?1), on the post‐thaw survival of sperm (triplicate set of six fish) were evaluated. Then, sperm was diluted in 1:1 mT extender with 10 mg mL^?1 BSA with selected cryoprotectants (15% MeOH and 10% DMSO) for 5 min. After a month of storage in liquid nitrogen, post‐thawed sperm motility; fertilization and hatching rate and viability of derived larvae were measured (Exp.3). Evaluation of cryoprotectants efficiency showed that MeOH 15% and DMSO 10% were suitable for cryopreservation of Persian sturgeon sperm. Gly and EG resulted in very low post‐thaw motility rates even at lowest concentration. No significant difference was observed among the four different equilibration times (0, 5, 10, 20 min) (P > 0.05) although higher equilibration times than 20 min resulted low post‐thaw motility (P < 0.05). The motility of frozen–thawed sperm did not significantly change when dilution ratio was increased from 1:0.5 to 1:3 (P > 0.05). However, higher dilution ratios (1:5 and 1:10) reduced the percentage of motile sperm. Supplementation of the cryoprotectant solution with 10 mg mL^?1 BSA significantly improved post‐thaw motility (P < 0.05), but ascorbic acid did not improve post‐thaw motility (P > 0.05). The results of experiment 3 showed that the highest fertilization (30.2 ± 5.75) and hatching rates (28.2 ± 5.25) were observed when samples were frozen with 15% MeOH (P > 0.05). Our study indicates that the use of mT extender consisting of 10 mg mL^?1 BSA in 15% MeOH diluted with sperm at 1:1 ratio for 5 min can be recommended cryopreservation method for Persian sturgeon sperm.