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Buffalo is an important farm animal species in South and South-east Asian countries. Cryopreservation allows long-term storage of somatic cells, which can be made available to research communities. This study aimed to 1) establish and cryopreserve somatic cells from elite buffaloes, and 2) share stored somatic cells and their associated data with researchers. To achieve these targets, somatic cells were established successfully from tail-skin biopsies of 17 buffaloes. The informative data such as buffalo details (breed, date of birth, sex, and age at the time of tissue biopsy collection, and production traits), the number of cryovials stored, and freezing dates were recorded in an electronic file and a printed inventory record. The established somatic cells were flat, spindle-shaped morphology, and expressed vimentin (a fibroblast-like cell type marker) and the negative expression of cytokeratin-18 (an epithelial cell type marker). Altogether, we cryopreserved 970 cryovials (0.1 million cells per vial) from two buffalo breeds, namely Murrah and Nili-Ravi (at least 45 cryovials per animal), for cryobanking. Somatic cell nuclear transfer (SCNT) experiments demonstrated the utility of cryopreserved cells to produce cloned buffaloes. Importantly, these cryopreserved somatic cells are made available to scientific communities. This study encourages the cryopreservation of somatic cells of elite farm animals for their utilization in cell-based research.

High temperature is one of the leading factors for decline reproductive performance in livestock and other species as a result of heat stress causing severe economic losses. The embryonic death causes due to heat stress is having multifactorial mechanism in live animal. Heat stress could influence reproductive physiology through modulating blood flow to the reproductive tract, ovarian steroid concentrations and patterns of follicular development. It is difficult for embryos to survive in the increase an oviductal and uterine temperature which is coincident with heat stress. In vitro culture of embryos at high temperatures has been reported to affect embryonic development. Similarly, increased in vitroculture temperatures can compromise oocyte activity and reduce fertilization rate. Studies have demonstrated that there were lethal effects of heat shock on in vitro cultured embryos of cattle at 41.0 – 43.0°C. However, these experimental temperatures are higher than those generally experienced by heat-stressed cows which ultimately reduced their fertility. Furthermore, a lot of research have been conducted in livestock species all indicating that exposure to high temperature is detrimental to oocytes and embryonic developmental processes as it leads to cell damage and may interfere with oocyte maturation, and fertilization process. It concludes that the longer exposure of oocytes and embryos to high temperatures causes more damage to oocytes and embryos.

This study was aimed at determining the presence of five major yield responsive genes shown to be among key determinants of rice yield in ten rice cultivars grown in Nigeria using candidate gene SSRs markers (Cg-SSRs). DNAs were extracted from young leaf samples using optimized cetyltrimethylammonium bromide (CTAB)-extraction method. An established gene sequences for high yielding rice Nipponbare cultivar for the five genes were retrieved from NCBI database and used to mine for SSRS using SSR identification tool. A pair of primer were designed manually from the nucleotide sequences flanking the selected SSRS for each gene. The designed primers were used to screen for the presence of the genes. Three pair of primers (for Gif1, Gn1a and Gw2) were able to amplify the targeted regions. The remaining two (for Gs3 and Dep1) did not amplify even the positive control. Two representative polymerase chain reaction (PCR)-products for the PCR-positive genes were sequenced and compared with established gene sequences of high yielding cultivars on NCBI database and their percentage sequence identities were determined. Phylogenetic trees (dendrograms) were generated using neighbor joining method. Overall, all the ten cultivars have both Gif1 and Gn1a genes while only eight cultivars have Gw2 gene. Gs3 and Dep1 need further optimization of PCR-conditions. Key words: Polymerase chain reaction, SSRs markers, primer.

As the coronavirus disease 2019 (COVID-19) pandemic continues to rise and second waves are reported in some countries, serological test kits and strips are being considered to scale up an adequate laboratory response. This study provides an update on the kinetics of humoral immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and performance characteristics of serological protocols (lateral flow assay [LFA], chemiluminescence immunoassay [CLIA] and ELISA) used for evaluations of recent and past SARS-CoV-2 infection. A thorough and comprehensive review of suitable and eligible full-text articles was performed on PubMed, Scopus, Web of Science, Wordometer and medRxiv from 10 January to 16 July 2020. These articles were searched using the Medical Subject Headings terms ‘COVID-19’, ‘Serological assay’, ‘Laboratory Diagnosis’, ‘Performance characteristics’, ‘POCT’, ‘LFA’, ‘CLIA’, ‘ELISA’ and ‘SARS-CoV-2’. Data from original research articles on SARS-CoV-2 antibody detection ≥second day postinfection were included in this study. In total, there were 7938 published articles on humoral immune response and laboratory diagnosis of COVID-19. Of these, 74 were included in this study. The detection, peak and decline period of blood anti-SARS-CoV-2 IgM, IgG and total antibodies for point-of-care testing (POCT), ELISA and CLIA vary widely. The most promising of these assays for POCT detected anti-SARS-CoV-2 at day 3 postinfection and peaked on the 15th day; ELISA products detected anti-SARS-CoV-2 IgM and IgG at days 2 and 6 then peaked on the eighth day; and the most promising CLIA product detected anti-SARS-CoV-2 at day 1 and peaked on the 30th day. The most promising LFA, ELISA and CLIA that had the best performance characteristics were those targeting total SARS-CoV-2 antibodies followed by those targeting anti-SARS-CoV-2 IgG then IgM. Essentially, the CLIA-based SARS-CoV-2 tests had the best performance characteristics, followed by ELISA then POCT. Given the varied performance characteristics of all the serological assays, there is a need to continuously improve their detection thresholds, as well as to monitor and re-evaluate their performances to assure their significance and applicability for COVID-19 clinical and epidemiological purposes.

OBJECTIVES West Nile virus (WNV) is a re-emerging mosquito-borne viral infection. This study investigated the pooled prevalence pattern and risk factors of WNV infection among humans and animals in Nigeria. METHODS A systematic review was conducted of eligible studies published in PubMed, Scopus, Google Scholar, and Web of Science from January 1, 1950 to August 30, 2020. Peer-reviewed cross-sectional studies describing WNV infections in humans and animals were systematically reviewed. Heterogeneity was assessed using the Cochrane Q statistic. RESULTS Eighteen out of 432 available search output were eligible and included for this study. Of which 13 and 5 were WNV studies on humans and animals, respectively. Although 61.5% of the human studies had a low risk of bias, they all had high heterogeneity. The South West geopolitical zone of Nigeria had the highest pooled prevalence of anti-WNV immunoglobulin M (IgM; 7.8% in humans). The pooled seroprevalence of anti-WNV IgM and immunoglobulin G (IgG) was 7.1% (95% confidence interval [CI], 5.9 to 8.3) and 76.5% (95% CI, 74.0 to 78.8), respectively. The WNV RNA prevalence was 1.9% (95% CI, 1.4 to 2.9), while 14.3% (95% CI, 12.9 to 15.8) had WNV-neutralizing antibodies. In animals, the pooled seroprevalence of anti-WNV IgM and IgG was 90.3% (95% CI, 84.3 to 94.6) and 3.5% (95% CI, 1.9 to 5.8), respectively, while 20.0% (95% CI, 12.9 to 21.4) had WNV-neutralizing antibodies. Age (odds ratio [OR], 3.73; 95% CI, 1.87 to 7.45; p

International audience; Background: Alkaline proteases is the important group of enzymes having numerous industrial applications including dairy food formulations.Objectives: The current study deals with the purification and characterization of an alkaline serine protease produced by Geotrichum candidum QAUGC01, isolated from indigenous fermented milk product, Dahi.Material and Methods: In total twelve G. candidum strains were screened for their proteolytic activity by using standard protease assay. The protease production from G. candidum QAUGC01 was optimized by varying physio-chemical conditions. The protease was purified by using two-step method: ammonium sulfate precipitation and gel filtration chromatography. Protease was further characterized by studying various parameter like temperature, pH, modulators, metal ions and organic solvent. A thermodynamic study was also carried out to explore the half-life of protease.Results: The G. candidum grew profusely at 25 °C and at an initial pH of 4.0 for 72 h of incubation producing 26.21 U/mlmaximum extracellular protease. Protease revealed that Vmax and Km was 26.25 U.ml-1.min-1 and 0.05 mg.mL-1, respectively using casein as substrate. The enzyme was stable at a temperature range (25-45 ºC) and pH (8-9). Residual enzyme activity was strongly inhibited in the presence of PMSF (7.5%). The protease could hydrolyze proteinaceous substrates, casein (98%) and BSA (95%). The thermodynamic studies explored that the half-life of the enzyme that was 106.62 min, 38.72 min and 15.71 min at 50, 60 and 70 ºC, respectively.Conclusions: Purified protease from G. candidum GCQAU01 is an ideal candidate for industrial application.

Simple Summary In this study, a chronic, seven-week study of the effect of technical grade glyphosate on the toxicity parameters of crossbred red tilapia (O. niloticus × O. mossambicus) was carried out. The results show that the bodyweight index was the most sensitive toxicity parameter wherein a reduction in body weight was observed at 25 mg/L of glyphosate. Negative correlations between the glyphosate concentration and toxicity parameters such as specific growth rate (SGR), hepato-somatic index (HIS), and gonado-somatic index (GSI) were observed. The fish condition factor and feed conversion ratio were found to be unaffected at the highest glyphosate concentration tested (150 mg/L). Abstract The development of glyphosate-resistant genetically modified organisms (GMO) has increased the use of herbicide glyphosate by several magnitudes in recent years. It is now the most commonly used pesticide globally that affects aquatic habitats, especially fish. This study aims to add new knowledge on the effect of technical grade glyphosate on several toxicity parameters and to identify the most effective parameter in predicting technical grade glyphosate chronic toxicity (seven weeks) to fish, especially Malaysia’s heavily farmed red tilapia. The results show that a relatively high concentration of technical grade glyphosate is needed to induce significant changes in all tested parameters. However, the results also indicate that the bodyweight index is the most sensitive toxicity parameter in that a reduction in body weight was observed at 25 mg/L of glyphosate. Negative correlations between the glyphosate concentration and toxicity parameters such as specific growth rate (SGR), hepato-somatic index (HIS), and gonado-somatic index (GSI) were observed. The fish condition factor and feed conversion ratio were found not to be affected at the highest glyphosate concentration tested (150 mg/L). To conclude, crossbred red tilapia (O. niloticus × O. mossambicus) is one potential species for evaluating the toxic effects of technical grade glyphosate on fish.

Diversification of histone variants is marked by the acquisition of distinct motifs and functional properties through convergent evolution.1-4 H2A variants are distinguished by specific C-terminal motifs and tend to be segregated within defined domains of the genome.5,6 Whether evolution of these motifs pre-dated the evolution of segregation mechanisms or vice versa has remained unclear. A suitable model to address this question is the variant H2A.W, which evolved in plants through acquisition of a KSPK motif7 and is tightly associated with heterochromatin.4 We used fission yeast, where chromatin is naturally devoid of H2A.W, to study the impact of engineered chimeras combining yeast H2A with the KSPK motif. Biochemical assays showed that the KSPK motif conferred nucleosomes with specific properties. Despite uniform incorporation of the engineered H2A chimeras in the yeast genome, the KSPK motif specifically affected heterochromatin composition and function. We conclude that the KSPK motif promotes chromatin properties in yeast that are comparable to the properties and function of H2A.W in plant heterochromatin. We propose that the selection of functional motifs confer histone variants with properties that impact primarily a specific chromatin state. The association between a new histone variant and a preferred chromatin state can thus provide a setting for the evolution of mechanisms that segregate the new variant to this state, thereby enhancing the impact of the selected properties of the variant on genome activity.

Citrus limon (L) leaf and stem used traditionally for the treatment of diseases and infections was phytochemically and antimicrobially screened. And also Thin layer Chromatography (TLC) was carried out on the crude methanol extracts. The phytoconstituents were qualitatively determined. The phytochemical screening of the crude methanol extract revealed the presence of saponins, terpenes, flavonoids, steroids, tannins, carbohydrates, anthraquinones, alkaloids, volatile oils and glycosides. The result of the antimicrobial screening of the crude methanol extract of the leaf showed potent activity against Staphylococcus aureus, candida albicans and bacillus subtillis. However, the crude methanol extract of the stem only showed activity against Staphylococcus aureus. TLC analysis of the of the crude methanol extract of the leaf showed three (3) spots while that of the stem revealed two (2) spots.

The major histocompatibility complex (MHC) is a polymorphic gene cluster of about 150 genes, present in all vertebrates. Many of these genes contribute to immunity. Particularly, MHC‐encoded class I and class II molecules, which are typically highly polymorphic and polygenic, are central in defining the specificity of the adaptive immune response. Among the diversity of genes associated with disease resistance, MHC genes are particularly interesting as they are associated with resistance and susceptibility to a wide range of diseases, some of which produce important economic losses in livestock. Enzootic bovine leukosis is an infectious disease caused by the retrovirus bovine leukemia virus (BLV), with an important economic impact, mainly in dairy herds. In this chapter, MHC‐associated genetic resistance to BLV is revised. Certain alleles of the bovine MHC (BoLA) class II locus have been found strongly associated with resistance to viral dissemination. Genetic selection of resistant animals emerges as a natural strategy for the control of infectious diseases, especially when there is no other alternative of control or prevention, as vaccines. Founded on this knowledge, a BLV control program based on selection of genetically resistant cattle was designed. The proof of concept indicates that this strategy is feasible to implement in dairy herds. Fil: Gutiérrez, Silvina Elena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina Fil: Esteban, Eduardo Néstor. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina Fil: Lützelschwab, Claudia María. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina Fil: Juliarena, Marcela Alicia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tandil. Centro de Investigación Veterinaria de Tandil. Universidad Nacional del Centro de la Provincia de Buenos Aires. Centro de Investigación Veterinaria de Tandil. Provincia de Buenos Aires. Gobernación. Comision de Investigaciones Científicas. Centro de Investigación Veterinaria de Tandil; Argentina