Carrier rate of the c.235G>C mutation in the bovine isoleucyl-tRNA synthetase (IARS) gene of Japanese Black cows at Kagoshima prefecture, Japan,and analysis of the metabolic profiling and reproductive performance of heterozygous cows

Bovine isoleucyl-tRNA synthetase (IARS) disorder, a major cause of weak calf syndrome, is caused by a homozygous missense (c.235G>C) mutation in the bovine IARS gene of Japanese Black (JB) cattle, which was identified in 2013. However, the extent to which the carrier rate has changed at Kagoshima prefecture, Japan, and whether the carrier status is associated with any clinical or reproductive problems, have yet to be ascertained. In this study, using a real-time polymerase chain reaction-based genotyping assay, we determined the carrier rate in a regional JB cow population at Kagoshima prefecture. Comparative analyses were performed on the metabolic profile test (MPT) results and reproductive performance data obtained for heterozygous carrier and homozygous wild-type cows. In 2009 and 2018, DNA samples were collected from 130 and 462 clinically healthy JB cows, respectively, in Kagoshima prefecture. MPT results and reproductive performance data were evaluated for 62 cows, comprising four heterozygous carriers and 58 wild-type cows. Genotyping revealed that the carrier rate was 6.9% in 2009 and 1.5% in 2018, the difference of which was statistically significant (P<0.005). There were no statistically significant differences between the carrier and wild-type cows with respect to either MPT results or reproductive performance, indicating that the carrier cows have necessary IARS activity to maintain minimal health and reproductive potential.     

New technique of sex preselection for increasing female ratio in boar sperm model

In this study, we tried to optimize the porcine semen extender conditions to maximize the differences between live X chromosome-bearing (X) spermatozoa and to Y chromosome-bearing (Y) spermatozoa without a decline in the fertility rate at different pH conditions during storage. We observed the viability of X and Y boar spermatozoa in acidic (pH 6.2), original (pH 7.2), and alkaline condition (pH 8.2) for 5 days to investigate the effect of storage conditions on the X to Y spermatozoa ratio. The functional parameters of spermatozoa were also examined to evaluate sperm quality. Sperm motility was preserved at pH 7.2 and pH 6.2 for 3 days, while sperm motility at pH 8.2 decreased significantly after 2 days. Non-capacitated spermatozoa increased while capacitated spermatozoa decreased during storage. Sperm viability decreased significantly duration-dependent under all pH conditions, but there was no significant difference during storage at pH 6.2 and 7.2. The X: Y ratio of live spermatozoa in acidic condition was maximized (1.2:1) without affecting the sperm function and fertility-related protein expression after 2 days compared to original conditions. Moreover, insemination of sows using acidic extender increased the number of female pups on days 1 and 2 of preservation. These results indicate that the production of female offspring may increase when acidic BTS is used for 2 days without affecting the success rate of AI. Above all, this method is simple and economical compared to other methods.     

Comparison of growth,survivorship, seed production and shedding of eight weed species in a wheat crop in Western Australia

Development of integrated weed management strategies is dependent on a thorough knowledge of the demography of individual species. The current research established eight winter or summer weed species in a winter annual wheat cropping system at Wongan Hills, Western Australia, and investigated emergence of the first cohort of each species, survivorship, plant size, seed production and seed shedding over three years (2016–2019). The winter weeds Bromus diandrus and Lolium rigidum emerged at the same time as the wheat crop, and the initial cohort of marked plants had 100% survival to seed production in each year. By comparison, other winter weed species like Hordeum leporinum, Rumex hypogaeus, Sonchus oleraceus and Polygonum aviculare frequently emerged later than the crop and had a lower percentage of plants surviving to seed production. However, individual S. oleraceus and P. aviculare plants had the greatest seed production compared to other species. All winter weeds had variable patterns of seed shedding between years, with the exception of L. rigidum. Summer weed species emerged at the same time, but plants in the initial cohort of each species did not always survive to produce seed. The early emergence and high survivorship of B. diandrus indicates high competitive ability, but shedding commenced at a similar time to L. rigidum and harvest weed seed control may be a viable control method for this species.     

Comparison of dot blot and PCR diagnostic techniques for detection of white spot syndrome virus in different tissues of Penaeus monodon

Comparison of PCR and dot blot diagnostic techniques for detection of white spot syndrome virus (WSSV) was made on different tissues of infected Penaeus monodon including eye stalk, eye stalk with eye, gills, cuticle, pleopod, periopods, uropods and telson. Dot blots of crude DNA extracted from infected tissue samples showed positive reactions with all the samples; however, the sensitivity of the dot blot was reduced with the purification of DNA samples extracted from pleopod, telson and uropod. PCR was found to be more sensitive when compared to dot blot. Both crude DNA and purified DNA samples extracted from all the tissues except for eye stalk with eye showed single step nested PCR positive reaction. The amplification of all or either of the three bands of 941 bp, 525 bp and 204 bp size varied with the tissues analysed. The severity of infection assessed by PCR amplification was found to be maximum in cuticle and telson followed by gill. Other tissues such as eye stalk, pleopod, periopods and uropod were observed to have mild infection. The maximum intensity of the PCR product was for the smallest amplified product of 204 bp followed by 525 bp and the weakest intensity was observed for the 941 bp size. The limitation of PCR due to inhibiting factors present in tissues could be overcome with the use of dot blot which gave positive reaction from the DNA extracted from eye stalk including the eye but yielded no amplification by PCR.     

Prostanoids-induced dispersion in the melanophores of a carp Labeo rohita (Ham.)

Effects of three prostaglandins (i.e., prostanoids) and one of its precursors, arachidonic acid, were examined on the melanophores of the fish Labeo rohita (Ham.). PGE₁, PGE₂, PGF_2α and arachidonic acid elicit a concentration-related dispersion in the fish melanophores. In vitro analysis of melanophores was performed through incubation of the isolated fish scales in different agonists and antagonists solutions. Dispersal effect of prostanoids may be mediated directly through the typical receptors or indirectly through release of neurotransmitter substance(s) from the melanophore nerve endings. Denervation of fish melanophores rendered them insensitive to prostanoid (PGF_2α). Propranolol and verapamil completely inhibited the dispersal effects of PGF_2α; theophylline and indomethacine blocked the effects of higher concentrations of PGF_2α. During dispersing influence of PGF_2α, a free flux of Ca²⁺ ions was required and the indirectly released substance(s) from melanophore nerve endings would be the catecholamines of adrenergic and purinergic in nature.     

Sodium alginate supplementation modulates gut microbiota,health parameters,growth performance and growth‐related gene expression in Malaysian Mahseer Tor tambroides

This study investigated the effects of sodium alginate supplementation on gut microbiota composition, health parameters, growth performances and growth‐related gene expression of Malaysian mahseer. Five test diets were formulated by supplementing 0%, 0.1%, 0.2%, 0.4% and 0.8% sodium alginate. Triplicate groups of juvenile Tor tambroides (2.19 ± 0.05 g) were stocked in 15 aquaria (20 individuals per aquarium) and fed at 3.0% body weight per day for 60 days. PCoA and UPGMA analysis showed that gut bacterial community were more convergence in higher sodium alginate‐supplemented diets. The percentage of Porphyromonadaceae, Bacteroides, Plesiomonas and Shewanella were substantially higher and Aeromonas, Entomoplasmatales and Prevotellaceae were drastically lower in higher sodium alginate (0.2%–0.8%) diets. Sodium alginate supplementation (≥0.2%) significantly improved the haematocrit value and respiratory burst activity of T. tambroides. Growth performances and feed utilization were significantly higher in 0.2%–0.4% sodium alginate‐supplemented diets. The increased growth rate of T. tambroides was governed by both hyperplastic and hypertrophic muscle growth. Real‐time PCR data demonstrated that most of the growth‐related genes were significantly upregulated in 0.2%–0.4% sodium alginate‐supplemented diet. Finally, it can be concluded that sodium alginate should be supplemented at 2 g/kg in practical fish feed formulation.     

Effect of high inclusion of rendered animal by‐product ingredients on growth,digestibility and economic performances in climbing perch Anabas testudineus

A 70‐day growth trail was conducted to investigate the effects of inclusion of high levels of meat and bone meal (MBM) and protein concentrate (PC) on growth, digestibility and economic performances of climbing perch, Anabas testudineus. Four isonitrogenous diets were formulated by lowering the level of dietary fishmeal protein at 0 (D₁, control), 70 (D₂), 85 (D₃) and 100% (D₄) with a mixture of MBM and PC protein (1:1). Triplicate groups of 300 fish (mean weight of 0.80 g) stocked in each 40 m² pond and fed the respective test diets. A digestibility trial was conducted after the growth trial in indoor glass aquarium. The result showed that growth parameters were significantly decreased (P < 0.05) with fishmeal replacement levels. However, significant differences were not found in feed conversion ratio and survival of fish. No difference was also found in protein efficiency ratio among D1, D2 and D3. Similar to growth parameters, total fish production was highest in D1, intermediate in D2 and D3; and lowest in D4. Apparent digestibility coefficients of dry matter, protein and lipid were highest (P < 0.05) in D1 and lowest in D4. The economic analysis revealed that the benefit cost ratio was ranked by D₃ (1.81), D₂ (1.71), D₁ (1.66) and D₄ (1.46) respectively. Upon considering the overall performances and unavailability of finite protein sources, it can be concluded that 70–85% fishmeal could be replaced with a mixture of MBM and PC (1:1) in practical diets for climbing perch.     

Can fermented soybean meal and squid by‐product blend be used as fishmeal replacements for Japanese flounder (Paralichthys olivaceus)?

An experiment was carried out to evaluate fermented soybean meal and squid by‐product blend (1:1) (FP) as replacement of fishmeal (FM) for Japanese flounder, Paralichthys olivaceus. Five isocaloric (19 kJ g^?1) diets were prepared by replacing 0 (FP0), 12 (FP12), 24 (FP24), 36 (FP36) and 48% (FP48) FM protein with FP. Triplicate groups of juveniles (mean weight of 3.9 g) were delivered the test diets for 8 weeks in a flow‐through sea water system. The results showed that there were no significant differences (P > 0.05) among the growth rates of fish fed FP0, FP12, FP24 and FP36 diets. Growth and nutrient utilization parameters were significantly reduced in fish fed FP48 diet. Although, whole body proximate composition of fish were not significantly affected by the dietary treatments compared to the control; methionine and phenylalanine contents were significantly decreased in FP48 group. Protein retention was also significantly decreased in the similar group of fish. Dietary treatments did not alter most of the plasma metabolites, while some of the health parameters were improved in the replacement groups. Results suggested that FP is a potential candidate for alternative protein ingredient in aquafeed and can replace 36% FM protein in the diet of Japanese flounder.