<div>Background: External skeletal fixator (ESF) has been recommended as biological method to stabilize open fracture as it does not involves the wound site, besides protecting adjacent soft tissue structures. Considering the advantages of ESF, this study was envisaged with an objective to study the outcome of open fractures of radius-ulna and tibia-fibula stabilized with linear ESF in dogs. 
 
Methods: This study included 17 dogs suffering from open long bone fractures of radius-ulna and tibia-fibula, divided into three groups; linear ESF type I or II (group I, n=8) and type III (group II, n=3). In group III (n=6), open fractures were initially treated using conservative methods i.e management of open wound till complete healing followed by delayed internal fixation using intramedullary pin or bone plating. 
 
Result: Majority of the open fractures of radius-ulna and tibia-fibula were transverse (64.71%), involving distal third diaphysis (47.06%) in heavy weight (54.55%), 1-3 year old (64.71%), non-descript (35.29%), male (88.23%) dogs due to automobile accidents (58.82%). Neutrophilic (81.47&plusmn;4.45%) leukocytosis (24610&plusmn;2461.88 per cumm) and elevated levels of alkaline phosphatase and creatine kinase were observed. Staphylococcus spp. was the common isolated bacteria sensitive to Gentamicin, Cefotaxime, Tobramycin, Amikacin and Neomycin. High per cent fractures (83.33%) in group III had acceptable functional outcome as compared to fractures treated with linear ESF (55.55%) in groups I and II. Osteomyelitis due to pin tract infections was the major complication (8/11; 72.7%) recorded in ESF groups (Group I and II) that lead to delayed and/or nonunion. Weight bearing score improved markedly after removal of the ESF frame. In conclusions, immediate fixation of the long bone fracture using linear ESF in the presence of open contaminated / infected wounds leads to unacceptable outcome. Poor animal compliance, pin tract discharge, osteomyelitis, delayed union and nonunion are major complications associated with linear ESF. Conservative management of the open fracture wound using standard treatment for 2-3 weeks till complete wound healing followed by internal fixation with intramedullary pin or bone plate is recommended for open long bone fractures in dogs.</div>

<div>Outcome of Open Long Bone Fractures in Dogs Stabilized with Linear External Skeletal Fixator</div>

<div>Background: The metallic ESF systems are not easily available in the field and have fixed frames leading to less versatility in size and direction. Free forms of external skeletal fixators (acrylic and epoxy-pin) have advantages as they can be customized. Both acrylic and epoxy-pin ESF systems have been evaluated biomechanically in earlier studies; in the present study both are compared clinically. 
 
Methods: Twenty four dogs presented with 27 fractures of long bones, were randomly divided into 4 groups, according to the bones involved, group A was divided into A-I (radius-ulna) and A-II (tibia-fibula) sub-groups and group B was divided into B-I (radius-ulna) and B-II (tibia-fibula) sub-groups. 
 
Result: The mean&plusmn;SE fracture healing time in animals was 48.85&plusmn;2.56 days with no significant difference between two groups.The early presented cases and the ones with less soft tissue trauma showed better gait scores during the follow up period. Both acrylic and epoxy-pin fixators provided stable fixation and the technique can be practiced at field conditions with minimal facilities. Epoxy-pin ESF owing to better handling characteristics of epoxy resin is easier to construct than the acrylic ESF.</div>

<div>Acrylic and Epoxy-pin External Skeletal Fixation Systems for Fracture Management in Dogs</div>

Background: To explore the impact of agronomic practices on nodulation, nitrogenase activity and biological nitrogen fixation in hybrid pigeonpea, a field investigation was under taken at BW5 block ICRISAT development center (IDC), International Crops Research Institute for Semi-Arid Tropics (ICRISAT), Patancheru, Hyderabad, during kharif, 2021 and 2022. 
 
Methods: The seedlings were raised in portrays in advance to onset of monsoon during last week of May (28th May in 2021 kharif and 30th May in 2022 kharif) in both the years by filling portrays with cocopeat and vermicompost in 1:1 ratio. Transplanting of 24 days and 25 days old seedlings were done on 20th June in 2021 kharif and 23th June in 2022 kharif, respectively, on the same day of dibbling in the main field at 100 cm &times; 100 cm ,120 cm &times;&nbsp;120 cm square geometry and 150 cm &times;&nbsp;60 cm. Nutrient management treatments were imposed one week after sowing. 
 
Result: Among the planting methods higher nodule number and nitrogenase activity with transplanting compared to dibbling. 120 cm &times;&nbsp;120 cm found higher nodule number plant-1 and nitrogenase activity but biological nitrogen fixation was higher with 100 cm &times;&nbsp;100 cm ha-1 basis. With respect on nutrient management practices, control performed well in terms of higher nitrogenase activity and biological nitrogen fixation and on par with integrated approach of 100% soil test based NPK + vermicompost + PSB + seed treatment with Rhizobium. So, with transplanting method of establishment, 100 &acute; 100 cm square plant geometry and 100% soil test based NPK + vermicompost + PSB + seed treatment with Rhizobium was recommended agronomic practices for hybrid pigeonpea.

Hybrid Pigeonpea (ICPH 2740) Nodular Activity and Biological Nitrogen Fixation as Influenced by Agronomic Practices

<div>Background: Heart is the central organ of circulatory system that pumps blood into the blood vessels and performs many vital functions. Its development before birth must be studied to safeguard the animal from the occurrence and consequences of various developmental anomalies. The detailed morphometry of different internal parameters of heart especially in pre-natal sheep has not yet been reported. &nbsp;&nbsp; 
 
Methods: The foeti of sheep were divided into two age groups viz. mid prenatal (51-100 days) and late prenatal (101 to 150 days) with fifteen animals in each age group. The various internal parameters were recorded by using digital weighing machine, graduated measuring cylinder, digital Vernier&rsquo;s calliper, non-stretchable nylon thread and graduated scale. The data recorded was statistically analysed by independent t test with IBM SPSS 25.0 version software.&nbsp; 
 
Result: The average thickness of left ventricular wall of heart was measured as 2.85&plusmn;0.15 mm and 4.28&plusmn;0.07 mm in mid prenatal and late prenatal stages respectively with significant (p&le;0.01) differences between the ages. Similarly, the average diameter of left atrio-ventricular opening of heart was measured as 2.06&plusmn;0.27 mm and 6.08&plusmn;0.36 mm in mid prenatal and late prenatal stages respectively with significant (p&le;0.01) differences between the ages. The average length of septal papillary muscle in right ventricle was measured as 1.12&plusmn;0.10 mm and 3.29&plusmn;0.21 mm in mid prenatal and late prenatal stages respectively with significant (p&le;0.01) differences between the ages. Further, the average length of moderator band in heart was measured as 1.45&plusmn;0.11 mm and 4.34&plusmn;0.23 mm in mid prenatal and late prenatal stages respectively with significant (p&le;0.01) differences between the ages. Similarly, the average length of chordae tendinae in right ventricle of heart was measured as 0.97&plusmn;0.09 mm and 3.50&plusmn;0.34 mm in mid prenatal and late prenatal stages respectively with significant (p&le;0.01) differences between the ages.&nbsp;</div>

<div>Age Wise Internal Morphometrical Studies on the Heart of Pre-natal Non-descript Sheep</div>

Background: Alfalfa is a kind of high-quality legume grass with high crude protein content, good palatability, high grass yield and good nitrogen fixation capacity. Abiotic stresses have become the key factors restricting the yield and quality in alfalfa and other crops, such as drought, extreme changes in ambient temperature, floods, land salinization and acidification, while under the different abiotic stress conditions, gene expression levels will also change accordingly. The selection and using of appropriate reference genes are crucial for the accuracy of gene expression quantification. 
 
Methods: We chose the candidate internal reference genes of alfalfa from transcriptome sequence datasets (162 RNA-seq sequencing data) through comparative analysis. Finally, 10 candidate reference genes were selected. These candidate reference gene expressions were determined by RT-qPCR under three common abiotic stresses in production, such as alkali, drought and low temperature. The stability index of these candidate genes was calculated and evaluated correspondently using specific softwares and different algorithms, such as GeNorm, Normfinder, Bestkeeper, △Ct method and an online analysis tool RefFinder. 
 
Result: The results showed that all 10 pair candidate reference genes could be used in gene expression quantification except for GAPDH and Ms.33066 under low-temperature stress based on screening criteria. Under alkaline stress, the optimal reference gene is UBL-2a and the optimal combination of reference genes is GAPDH and UBL-2a; Under drought stress, the optimal reference gene is Rer1 and the optimal combination of reference genes is Ms.33066&nbsp;(some candidate reference genes haven&rsquo;t been annotated yet, using gene ID abbreviation number of Medicago sativa L. instead) and Actin; Under low-temperature stress, the optimal reference gene is Ms.99505 and the optimal combination of reference genes is Ms.65463, UBL-2a, Ms.99505 and Actin. Among all the samples, the optimal reference gene is MS.99505 and the optimal combination of reference genes is MS.073307, Rer1, MS.99505 and UBL-2a. GAPDH and Actin aren&rsquo;t the most appropriate reference genes of alfalfa under different abiotic stresses, the optimal reference gene and the optimal combination of reference genes under other abiotic stress need further validation. This paper provides scientific evidence for quantitative analysis of the genes of alfalfa.

Screening of Internal Reference Genes Analysis in Alfalfa under Different Abiotic Stress Conditions

<div style="text-align: justify;">Using a quartz sand culture, comparisons were made between the T5 homozygous transgenic lines (HTLs) that overexpress StP5CS (GenBank accession number: JN606861) and their wild-type (WT) host cultivar to examine the differences in the growth and development traits, the concentrations of proline in vegetable soybean [Glycine max (L.) Merrill] under salt stress conditions. Moreover, the relative expression levels of two glutamine synthetase-related genes (GmGS1&acirc;1, GmGS1&acirc;2), two nodulation-related genes (GmENOD40-1, GmENOD40-2), and one leghemoglobin gene (GmLba) were also measured. The purpose of this research was to provide a theoretical basis for elucidating the mechanisms of nodulation and nitrogen fixation in the roots of transgenic plants under salt stress conditions. Compared with WT plants, the plant height and seed weight per plant of T5 HTLs significantly increased, and the contents of proline in various tissues of T5 HTLs were also significantly elevated. Quantitative real-time PCR (qRT-PCR) analysis indicated that the expression levels of GmGS1&acirc;1, GmGS1&acirc;2, GmENOD40-1, &nbsp;GmENOD40-2, and GmLba were significantly increased in T5 HTLs under salt stress conditions. These results indicate that the overexpression of StP5CS in T5 HTLs enhanced growth, nodulation and nitrogen fixation in transgenic vegetable soybean under salt stress conditions.</div>

Effects of StP5CS gene overexpression on nodulation and nitrogen fixation of vegetable soybean under salt stress conditions

<div>Symbiotic N2 fixation is essential for the plant&rsquo;s growth because it can fix reactive nitrogen compounds in soil. However, all steps of this process can be hampered by several biotic and abiotic environmental factors. This study mainly focused on discussing the impacts of 12 major factors on this process by reviewing the significant numbers of research works. According to the information from these works, we found some significant physiological and genetic impacts caused by these factors like plasmid deletion, genomic mismanagement, abnormal molecular signals, toxicity, deficiency of minerals, deformation of rhizobial cells, protein denaturation, nucleic acid damage, acetylene reduction and nod factors limitation. Furthermore, expression of heat or acid shock proteins, internal buffering, genes spanning, extracellular immobilization, periplasmic allocation, change of lipopolysaccharides composition, intracellular accumulation of inorganic and organic solutes (Osmolyte) and activation of hydrogenase expression are shown by both micro and macro symbionts as a natural response to adapt to these stress conditions. Though the stress-tolerant strains like HR-3, HR-6, HR-10, HR-12, acta, actP, exoR, lpiA, actR, actS and phrR can be used to sense the external environment and make signals to change gene transcription during the adverse condition, the application of genetic engineering should be expanded more to promote the commercial inoculation by the production of novel stress-tolerant strains or modified genes of rhizobia and legumes.</div>

Factors Impacting Rhizobium-legume Symbiotic Nitrogen Fixation with the Physiological and Genetic Responses to Overcome the Adverse Conditions: A Review

<div style="text-align: justify;">Poultry farming is gaining strength in mitigating livelihood and nutritional security to the poor farmers which constitute 60% of the India&rsquo;s population. Thus eggs of superior quality is prerequisite for profitable marketing and for hatchability too. The present study was conducted on two improved varieties, &nbsp;Vanaraja (dual purpose type) and Gramapriya &nbsp;(the layer type) chicken developed at PDP, Hyderabad and their crosses on random mating. The genetic groups taken were Gramapriya (male) x Gramapriya (female),Vanaraja(male) x Vanaraja(female), Gramapriya(male) x Vanaraja(female)and Vanaraja (male) x Gramapriya(female). 6 males and 30 females were taken from each genetic group and maintained separately under deep litter system with a mating ratio of 1 male : 5 females. To study the genetic effect on external and internal egg quality traits, a total of more than 50 eggs were collected at random from each of 4 genetic groups upto 28 weeks of age. The average shell thickness obtained to be ranged from 0.36 to 0.39 mm. The average albumen and yolk indices were found to be ranged from 7.396 to 8.656 and 41.749 to 43.888 respectively. The average shell weight and percentage of shell weight observed to be ranged from 4.147 to 5.627 g and 17.266 to 18.003% respectively.</div>

Genetic impact on &nbsp;external and internal egg quality &nbsp;traits of Vanaraja and Gramapriya birds and their crosses in Bihar

<div>Background: Listeria monocytogenes is the only pathogenic bacterium of Listeria that causes disease to humans. The Dairy products, meat and other foods have been proved to be the transmission carriers of Listeria monocytogenes, endangering human health. Therefore, it is necessary to establish a fast, sensitive and specific fluorescent quantitative PCR method for detecting Listeria monocytogenes in milk based on internal reference. 
 
Methods: According to the genome sequence of Listeria monocytogenes published in Genbank, we screened specific target gene Hly gene, designed specific primers and probes, optimized the reaction system and added internal reference (IAC) to the system. This IAC was detected by TaqMan probes labeled with different fluorophore. 25-100 CFU/25 g artificially contaminated sample was added to evaluate the performance of reaction system. 
 
Result: The assay was could be used reliably for detection of Listeria monocytogeneswith a sensitivity of 10-3ng/uL. For the 10-fold dilutions bacteria DNA extracted by cooking water,the lowest detection limit was 4 x 102cfu/mL. But for the plasmid with hly, the lowest detection limit can reach 104 cfu/mL. The standard curve of hly and hly-IAC was established and the quantification was linear between Ct and template copy number (r2=0.999). While the initial sample amount of artificially contaminatedbacteria was 25 CFU/25 g (Fresh milk, milk powder, fermented milk), Listeria monocytogenes could be detected in 14 h. The Hly IAC fluorescent quantitative PCR detection method established in this study,and added internalreference tothe system, can accurately detect Listeria monocytogenesin fresh milk.At the same time, real-time monitoring of the PCR reaction process ensures the reliability of the results, which is suitable for rapid detection of large batches of samples.</div>

<div style="text-align:justify">&nbsp;</div>

Establishment of a Fluorescence Quantitative PCR Assay in Detection of Listeria monocytogenes based on Internal Reference

<div style="text-align: justify;">The StP5CS (GenBank accession number: JN606861) T6 homozygous transgenic lines (HTLs) of vegetable soybean [Glycine max (L.) Merrill] were grown using vermiculite pot culture to determine whether StP5CS overexpression would enhance nodulation and symbiotic nitrogen fixation (SNF) in two T6 HTLs (17W-1, 17W-2) under drought conditions. The growth performance, nodule development and seed weight of T6 HTLs were significantly better than those of wild type (WT) plants. The proline levels in various tissues of T6 HTLs were higher than WT plants. The concentrations of total ureide, total N, leghemoglobin (Lb) and the activity of glutamine synthetase (GS, EC 6.3.1.2) in the T6 HTLs were significantly increased. Moreover, the relative expression levels of five key nodulation- and SNF-associated genes (i.e., GmENOD40-1, GmENOD40-2, GmLba, GmGS1&acirc;1 and GmGS1&acirc;2) were significantly higher in T6 HTLs. In conclusion, overexpression of StP5CS enhances nodulation and SNF in transgenic vegetable soybean under drought stress conditions.</div>

<div style="text-align: justify;">Overexpression of the StP5CS gene promotes nodulation and nitrogen fixation in vegetable soybean under drought stress</div>