Effect of environmental factors on estrus synchronization and artificial insemination success in farmers flock in sheep under semi-arid tropical region

Environmental cues in the different seasons might influence the estrus and lambing percentages. The present study was conducted to assess the effect of environmental factors on estrus synchronization (ES) and artificial insemination success in sheep . During the period from 2012 to 2017, a total of 1,197 ewes of Kheri and Malpura breeds of 34 farms originating from 10 villages were synchronized for estrus and then fixed-time artificial insemination (FTAI) was done. Oestrous synchronization was done by intravaginal progesterone sponges and eCG protocol. Fixed-time cervical insemination was performed 48 and 56 hr after sponge removal in ewes exhibiting estrus, using liquid chilled semen containing 100 million sperm per dose of Patanwadi/Malpura rams. Mean sunshine hours, maximum and minimum ambient temperatures, temperature amplitude, mean relative humidity (RH), temperature-humidity index (THI), mean temperature during sponge-in to sponge-out—12-day period, sponge-out and next day, AI day and AI to next 15 days have been calculated. The estrus response and lambing percentage were higher (p < .05) when ES and FTAI were done during hot-humid with rainfall season. Successful (p < .05) ES occur when the minimum temperature was higher in cold-humid season, and the sunshine hour was higher in hot-humid with scanty rainfall season during sponge-in to sponge-out period. The success of the lambing percentage was higher (p < .05) when maximum temperature, minimum temperature, mean temperature and THI was higher on the day of AI and the next 15-day period, irrespective of the season. The success of FTAI indicates the adaptability of the local breed to tolerate the harsh climate of the hot semi-arid region without much hindrance in lambing percentages.     

Molecular identification of a new isolate of <Emphasis Type="Italic">Cucumber mosaic virus</Emphasis> subgroup II from basil (<Emphasis Type="Italic">Ocimum sanctum</Emphasis>) in India

Natural occurrence of mosaic disease was observed on basil (Ocimum sanctum L.) in Aligarh, U. P., India, during 2008. The disease could be transmitted by sap inoculations from naturally infected O. sanctum to O. sanctum and some test plant species. Cucumber mosaic virus (CMV) was detected by RT-PCR using coat protein gene specific primers of CMV (Acc. AM180922 & AM180923), which resulted in the expected size ~650 bp amplicon in infected samples. The amplicon was cloned, sequenced and data were deposited in GenBank Acc. EU600216. The sequence data analysis revealed 97–99% identities at both nucleotide and amino acid levels with the CMV strains of subgroup II reported worldwide. Based on the high sequence identities and close phylogenetic relationships with CMV subgroup II strains, the virus under study has been identified as a new isolate of CMV subgroup II and designated as CMV-Basil.     

Assessment of Genetic Variability in the Coding Sequence of Melatonin Receptor Gene (MTNR1A) in Tropical Arid Sheep Breeds of India

Seasonal behaviour in sheep, which varies in tropical and temperate environmental conditions, is a matter of study, because it can provide a clue to address the problem of seasonality in sheep. Melatonin receptor is the membrane‐bound G‐coupled receptor, sensing the message of photoperiodic cues thorough melatonin. Restriction fragment length polymorphism (RFLP) studies were carried out to assess the variability of gene at G612A and C606T SNPs in MTNR1A gene, which have been studied to be markers for out‐of‐season breeding. Allelic frequency distribution corresponded to higher frequency of GG and CC genotype, in tropical arid sheep breed in comparison with temperate region sheep breed. PCR amplification of MTNR1A gene of 30 animals was performed and single nucleotide polymorphisms (SNPs) identification was carried out using Lasergene software. Seven SNPs/mutations were identified, but most of them were synonymous, except the one G706A, leading to substitution of valine by isoleucine. Polyphen‐2 analysis of G706A mutation revealed that it is a benign mutation. Two important SNPs C426T and G555A, which were identified in temperate sheep breeds, could not be traced in Magra and Marwari breeds of sheep. Thus, the Magra and Marwari breeds of tropical, arid region demonstrated the presence of both polymorphic SNPs markers G612A and C606T, associated with out‐of‐season breeding. GG and CC genotypes were having a higher prevalence in the studied population.     

Growth and ion uptake of four Brassica species as affected by Na/Ca ratio in saline sand culture

The influence of supplemental Ca² in saline nutrient solutions on germination, growth, and ion uptake of four Brassica species, B. campestris L., B. carinata, A.Br., B. juncea (L.) Czern. and Coss., and B. napus L. was studied in sand culture. The addition of 11.25 mM CaC1² to nutrient solution containing 225 mM NaC1 improved percentage of germination of B. napus and B. juncea, but had no significant effect on the germination speed of the four species. There was no significant effect of additional amount of Ca² in the saline medium (150 mM NaC1) on the shoot biomass and seed yield of B. carinata and B. campestris. By contrast, shoot dry matter of B. napus and B. juncea increased significantly with the increased in Ca² concentration of the growth medium, but their seed yield remained unaffected. Decreased Ca² concentrations of the saline growth medium reduced percent oil content in B. carinata, B. juncea, and B. campestris. Increasing Na/Ca ratio of the external medium affected ion uptake differently in different species. In B. carinata, a relatively salt-tolerant species, the concentrations of Na⁺ and K⁺ in the shoots remained unaffected, but the C1 concentration was reduced linearly with the increase in external Na/Ca ratios. Root K⁺ and Ca²⁺ of the species decreased with the decrease in Ca²⁺ supply. In B. campestris increasing Na/Ca ratios of the saline medium had no effect on the concentrations of Na⁺, C1, and Ca²⁺ in the shoots and Na⁺, C1, and K⁺ in the roots. Only shoot K⁺ and root Ca²⁺ decreased consistently. In the highly salt-sensitive species, B. napus, the shoot Na⁺ was reduced by the addition of Ca²⁺ in the salt treatment, whereas the C1 and Ca²⁺ uptake was promoted by supplemental Ca²⁺. The root K⁺ concentrations decreased with the increase in Na/Ca. In B. juncea, which was similar to B. napus in biomass production, high Ca²⁺ concentration in the salt treatment reduced the shoot Na⁺ and root C1 concentrations and promoted the K⁺ uptake. Shoot Na/Ca and Na/K ratios were increased in B. napus and B. juncea at the highest Na/Ca ratio of the growth medium. Shoot K⁺ selectivity, S^K,Na (determined as molar ratio of K: Na in tissue to molar ratio of K:Na in external medium) of all species remained unaffected except for B. juncea in which it decreased significantly at the highest Na/Ca ratio. The root K⁺ selectivity increased in B. carinata.     

Effect of protection against hot climate on growth performance,physiological response and endocrine profile of growing lambs under semi-arid tropical environment

In the hot semi-arid tropical region, extreme summer is a major constraint in sheep production. The growth performance of growing lambs is impaired during the summer. Therefore, the present study was conducted to assess the effect of protection against hot climate on growth performance, physiological response, and endocrine profile of growing lambs under semi-arid tropical environment. All the data in the experiment were presented as mean ± SE. Thirty Malpura lambs with the age of 122.7 ± 6.05 days and body weight of 19.52 ± 0.42 kg were allotted into three groups, viz. G1 (control, lambs kept in open area under tree shade), G2 (lambs kept in conventional asbestos-roofed shed), and G3 (lambs kept in designed shed). The allotment was based on their initial body weight so that the mean body weight of each group was similar. In all three types of protection, lambs were provided with sufficient space to move and social interaction. In asbestos-roofed shed (G2), all the four sides from ground to the roof were fenced with strong galvanized iron chain link. But in G3, all the four sides were constructed in a manner that there was 0.13 m air space between inner and outer solid brick columns of the wall up to 1.37 m height, and rest up to the roof was fenced with bamboo splint net. The air space between the two columns of the wall was filled with sand, and it was kept in moist condition by continuous water drips which provide extra evaporative cooling. The designed shed was basically constructed with bamboo. Tree shade was assured under the natural shades of large trees. The shaded area was protected by wire fences. The experiment was conducted for 45 days during very high temperature (May–June). The lambs were provided with ad libitum green fodder, dry roughage, 300 g concentrate, and ad libitum drinking water. The respiration rate at morning and afternoon, pulse rate at morning and afternoon and rectal temperature at afternoon was significantly (P < 0.05) lower in lambs of G3 as compared to other groups. The final body weight did not differ significantly between the groups, but average daily gain (g/day) was higher in G3 (150.06 ± 11.13) as compared to the G1 (109.69 ± 18.10). The feed conversion rate (FCR) was significantly (P < 0.05) better in G3 (5.49 ± 0.08) as compared to G1 (7.65 ± 0.08) and G2 (6.60 ± 0.08). The hemoglobin level was significantly (P < 0.05) higher in G1. The endocrine profile did not vary significantly (P > 0.05) among the groups, but insulin-like growth factor-1 (IGF-1) was higher in G3. It can be concluded from this study the lambs provided with better protection during summer remained in more comfortable condition, showed better FCR, and although not statistically significant showed better growth performance.

     

Enhanced <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">tuberosi</Emphasis> Resistance in Transgenic Potato Expressing a Rice GLP Superoxide Dismutase Gene

Germin like proteins (GLPs) are a large group of related and ubiquitous plant proteins which are considered to be involved in different processes important for plant development and defense. Multiple functional copies of this gene family have been reported in a number of species (wheat, barley, rice, soybean mosses and liverwort), and their role is being evaluated by gene regulation studies and transgenic approaches. To analyze the role of a rice (Oryza sativa) root expressed germin like protein1 OsRGLP1, for its antifungal activity, transgenic potato plants were developed. These transgenic potato plants were molecularly characterized and biologically assessed after inoculation with Fusarium oxysporum f. sp. tuberosi. Functional analysis showed high accumulation of H₂O₂, increased Superoxide Dismutase (SOD) activity and no oxalate oxidase activity (OxO) in transgenics in comparison to nontransformed control. This increased SOD activity, resistance to heat and sensitivity to H₂O₂ suggest it is a Fe-like SOD. OsRGLP1 expression in potato plants exhibited enhanced resistance in comparison to nontransformed wild type plants suggesting its role in providing protection against Fusarium oxysporum f. sp. tuberosi through elevated SOD level. Overall, results suggest that OsRGLP1 is a candidate for the engineering of potato plants with increased fungal tolerance however, the greater height and tuber number was observed. This phenotype associated with the resistance needs to be evaluated to determine if this is a positive or negative feature.     

Physiological response of crossbred sheep under nutritional scarcity

Nutritional scarcity during summer and winter is a major constraint for sheep rearing in the semi-arid tropical region. In this region, a new crossbred sheep has been developed by the crossing of a native sheep breed Malpura with a breed of hot-humid coastal region breed Garole. A study was conducted for 9 weeks to assess the physiological response of crossbred (Garole x Malpura x Malpura, GMM) rams under nutritional scarcity in the semi-arid tropical region. Eighteen adult crossbred rams were randomly allocated into three groups of six animals each. The groups are G1 (control, ad libitum feeding), G2 (20% less than ad libitum feeding), and G3 (30% less than ad libitum feeding). The animals were stall-fed with a diet consisting of 70% roughage (Cenchrus ciliaris hay) and 30% concentrate feed. Body weight gain, average daily gain, and feed intake were significantly (P?<?0.05) lower in G2 and G3 as compared to G1 rams. Water intake was significantly (P?<?0.05) higher in G2 and G3. Physiological responses, blood biochemical and endocrine profile did not differ between the groups. The results indicate that the crossbred rams evolved through the crossing of the hot semi-arid tropical region breed with hot humid coastal region breed are able to maintain their body homeostasis despite nutritional scarcity although their body weight decreased.

     

Modulation of seminal plasma content in extended semen improves the quality attributes of ram spermatozoa following liquid preservation at 3–5°C

Seminal plasma (SP) is known to induce motility and capacitation in spermatozoa curtailing their lifespan when preserved. Hence, this study was conducted to examine the effects of removal of SP from sperm surface prior to liquid preservation either by high dilution (1/15) or by washing and the poststorage treatment with SP (15% and 25%, v/v) on the quality attributes of liquid‐preserved ram semen. Over the period of storage, the rapid motility (66.0% and 71.1% vs. 58.3%), straightness (87.1% and 82.1% vs. 79.4%), average path velocity (152.3 and 152.0 µm/s vs. 133.3 µm/s) and the straight‐line velocity (131.3 and 127.8 µm/s vs. 108.5 µm/s) were significantly (p < 0.05) higher in both the high‐dilution and wash groups as compared to the control (1/3 dilution). The functional membrane integrity (82.3% vs. 77.2%) and noncapacitated sperm count (65.0% vs. 58.7%) were also significantly (p < 0.05) higher in the high‐dilution and wash groups, respectively, as compared to the control. The poststorage treatment of sperm with SP significantly (p < 0.05) increased the functional membrane integrity (70.1% vs. 53.8%) and most of the motility attributes as compared to the control (without SP). In conclusion, both the removal of SP prior to liquid preservation and poststorage treatment with SP significantly improved the quality attributes of ram spermatozoa.     

Infiltrating Foxp3+ Regulatory T cells and Histopathological Features in Canine Classical and Spermatocytic Seminomas

In humans, regulatory T (T reg) cells are known to play a critical role in both the regulation of immune homoeostasis and the progression of cancer. However, there is little information about the identification, characterization and the function of T reg cells in canine tumours. We identified T reg cells in 28 canine seminoma samples using a Forkhead box P3 (Foxp3) antibody and investigated the relationship between T reg cell infiltration and histopathological features of classical and spermatocytic seminomas (SE and SS, respectively). The Foxp3 protein showed nuclear immunostaining in infiltrating lymphocytes, and Foxp3+ cells were diffused or focally distributed in seminoma tissues. Foxp3+ cells were frequently present in the SS histotype, in seminomas that showed no evidence of tumour cell invasion into the vessels and in seminomas showing a diffuse growth pattern with three cell types. Neither the SE/SS histotype nor the histopathological features of the tumour correlated with Foxp3+ cell counts. These results indicate that Foxp3+ T reg cells may be associated with a less malignant histological phenotype or may not play a critical role in the immune response of canine seminomas. Moreover, Foxp3+ T reg cells may be associated with SS seminoma, but further studies, involving a larger number of samples, are required to better understand whether these cells play a critical role in the immune response in canine seminomas. This is the first report to demonstrate the characteristics of T reg cell infiltration in canine seminoma.