Influence of co‐culture with denuded oocytes during in vitro maturation on fertilization and developmental competence of cumulus‐enclosed porcine oocytes in a defined system

Co‐culture of cumulus‐oocyte complexes (COCs) with denuded oocytes (DOs) during in vitro maturation (IVM) was reported to improve the developmental competence of oocytes via oocyte‐secreted factors in cattle. The aim of the present study was to investigate if addition of DOs during IVM can improve in vitro fertilization (IVF) and in vitro culture (IVC) results for oocytes in a defined in vitro production system in pigs. The maturation medium was porcine oocyte medium supplemented with gonadotropins, dbcAMP and β‐mercaptoethanol. Cumulus‐oocyte complexes were matured without DOs or with DOs in different ratios (9 COC, 9 COC+16 DO and 9 COC+36 DO). Consequently; oocytes were subjected to IVF as intact COCs or after denudation to examine if DO addition during IVM would affect cumulus or oocyte properties. After fertilization, penetration and normal fertilization rates of zygotes were not different between all tested groups irrespective of denudation before IVF. When zygotes were cultured for 6 days, no difference could be observed between all treatment groups in cleavage rate, blastocyst rate and cell number per blastocyst. In conclusion, irrespective of the ratio, co‐culture with DOs during IVM did not improve fertilization parameters and embryo development of cumulus‐enclosed porcine oocytes in a defined system.     

Use of next‐generation sequencing for the identification and characterization of Maize chlorotic mottle virus and Sugarcane mosaic virus causing maize lethal necrosis in Kenya

The diagnosis of novel unidentified viral plant diseases can be problematic, as the conventional methods such as real‐time PCR or ELISA may be too specific to a particular species or even strain of a virus, whilst alternatives such as electron microscopy (EM) or sap inoculation of indicator species do not usually give species level diagnosis. Next‐generation sequencing (NGS) offers an alternative solution where sequence is generated in a non‐specific fashion and identification is based on similarity searching against GenBank. The conventional and NGS techniques were applied to a damaging and apparently new disease of maize, which was first identified in Kenya in 2011. ELISA and TEM provided negative results, whilst inoculation of other cereal species identified the presence of an unidentified sap transmissible virus. RNA was purified from material showing symptoms and sequenced using a Roche 454 GS‐FLX+. Database searching of the resulting sequence identified the presence of Maize chlorotic mottle virus and Sugarcane mosaic virus, a combination previously reported to cause maize lethal necrosis disease. Over 90% of both viral genome sequences were obtained, allowing strain characterization and the development of specific real‐time PCR assays which were used to confirm the presence of the virus in material with symptoms from six different fields in two different regions of Kenya. The availability of these assays should aid the assessment of the disease and may be used for routine diagnosis. The work shows that next‐generation sequencing is a valuable investigational technique for rapidly identifying potential disease‐causing agents such as viruses.     

A novel pathovar of Xanthomonas axonopodis causes gumming of Guggal (Commiphora wightii)

Guggal (Commiphora wightii (Arnott) Bhandari comb. nov.) is a small tree which is tapped for medicinally important oleo?Cgum?Cresin. Naturally infected plant oozes oleo?Cgum?Cresin from its trunk and primary branches. However, in either case, the plant dies slowly after oozing. A bacterium was established to be responsible for these phenomena. Four isolates of this bacterium were characterised by biochemical tests, Biolog GN2 microplate reaction, rDNA sequencing, which suggested that the pathogen belonged to the genus Xanthomonas. However, phylogenetic analysis based on chaperone protein (dnaK) gene, TonB?Cdependent receptor (fyuA) gene, DNA gyrase B (gyrB) gene and RNA polymerase sigma factor (rpoD) gene sequences placed it as a member of X. axonopodis 9.2 rep?CPCR/DNA?CDNA homology cluster close to X. perforans, X. alfalfae and X. euvesicatoria. Further elucidation of phylogenetic position of the test strains was achieved from a gyrB based tree considering sequences from 71 representative strains. Test strains were confirmed to be members of X. axonopodis. These had very narrow infectivity limited to Commiphora spp. Hence, we propose a novel pathovar, X. axonopodis pv. commiphoreae pv. nov. as the cause of gum oozing in guggal. Pathotype is DXA 01 = CFBP 7580 = LMG 26789.     

Effect of Artificial Insemination Protocol and Dose of Frozen/Thawed Stallion Semen on Pregnancy Results in Mares

Deep intra‐uterine insemination is commonly accepted as a routine procedure for artificial insemination in horses. The motives and principles of deep insemination are well described, but the equipment used may differ. In this trial, the efficiency of two different insemination pipettes for deep intra‐uterine insemination in the mare was compared with insemination into the uterine body using commercially available frozen–thawed semen of two stallions of proven fertility. These inseminations were performed using two different doses. The semi‐flexible Minitube pipette was compared with a newly designed insemination device with a more flexible telescopic insemination catheter (Ghent device). The semi‐flexible Minitube pipette performed better than the newly designed insemination device with respect to pregnancy outcome (p = 0.008). The superiority of deep horn insemination over uterine body insemination was reflected by the better pregnancy rates obtained after deep insemination using the same low doses (30.6% better pregnancy rates) (p = 0.0123).     

Activin receptor type IIB in rohu (Labeo rohita): molecular characterization,tissue distribution and immunohistochemical localization during different stages of gonadal maturation

Fish Physiology and Biochemistry - Activin receptor type IIB (ActRIIB) is a transmembrane serine/threonine kinase receptor which plays a pivotal role in regulating the reproduction in vertebrates...     

Genetic divergence among a non-restorer collection of sorghum (Sorghum bicolor (L.) Moench) and its relationship with heterosis

Summary A genetic diversity analysis in a collection of 171 non-restorer lines of sorghum (Sorghum bicolor (L.) Moench) using D² technique and canonical variate analysis indicated that considerable variation in grain yield has been added to the collection by the addition of lines derived from random mating populations. The efficiency of D² and canonical variate techniques in distinguishing extremely diverse genotypes was confirmed. However, the two techniques showed weak correspondence in their clusters. The F₁ hybrids of 15 diverse lines exhibited no relationship between heterosis or per se performance of crosses and diversity in their parents. Therefore, traditional plant breeding methods are being advocated.Approved by ICRISAT as Journal Article no. 435.     

Use of the Sperm Quality Analyzer (SQA II‐C) for the Assessment of Dog Sperm Quality

In the present study, an automated system for sperm analysis, the Sperm Quality Analyzer (SQA II‐C), was tested as a potential tool for the assessment of dog sperm quality. In the first experiment the device displayed a good repeatability of measurements for semen of medium and high quality, as evidenced by a low coefficient of variance (CV; 0.08), whereas a high CV (0.46) was obtained for one dog with semen of inferior quality. In the second experiment, seven different sperm concentrations (25–300 ×10⁶/ml), obtained by dilutions in Hepes‐TALP medium were stored for 48 h at room temperature. A concentration dependent increase in sperm motility index (SMI) was shown, reaching a plateau at 150×10⁶ spermatozoa/ml. For all sperm concentrations, the SMI value decreased significantly after 24 h, indicating the importance of sperm motility for SMI values. For sperm concentrations lower than 150×10⁶/ml, highly significant correlations [r=0.80;p<0.05] were established between SMI values on one hand and sperm concentration, and semen parameters expressing the overall semen sample quality on the other hand (experiment 3) while non‐significant or low correlations were found between SMI values and other individual sperm parameters. In experiment 4, significantly high correlations (r=0.97) were found between mean SMI values and post‐thaw motility and progressive motility assessed subjectively. In conclusion, our study indicates that both motility and concentration largely influence SMI values and that the SQA II‐C saturates at 150×10⁶ fresh spermatozoa/ml. In our opinion, the SQA II‐C may be a useful and objective device to assess the post‐thaw motility of dog sperm.     

Pseudocercospora griseola,the causal agent of common bean angular leaf spot: Strain characterization and sensitivity to fungicides

Angular leaf spot (ALS), an important disease of common bean (Phaseolus vulgaris), is caused by the fungus Pseudocercospora griseola. This pathogen has a wide genetic variability and, therefore, poses a challenge to integrated disease management. The use of resistant cultivars is difficult; hence, the application of fungicides has been a common practice in common bean cultivation. P. griseola strains were morphophysiologically characterized and their sensitivity to common fungicides used to control ALS was studied. The strains were evaluated for sporulation capacity and a representative sample of 34 strains was bioassayed to determine their sensitivity to commercial concentrations of five fungicides, namely pyraclostrobin, mancozeb, pyraclostrobin + metconazole, chlorothalonil and tebuconazole. Another sample of 29 strains was studied for conidial germination and dimensions. Sporulation capacity ranged from 0.88 to 27.67 × 10⁴ conidia/ml and germination percentage ranged from 39% to 72%. The large differences among strains suggest a wide genetic variability among the strains. A wide variability in aggressiveness of P. griseola was observed, which has consequences for breeding programmes aimed at resistance. The behaviour of pathogen strains differed in every fungicide evaluated, even in a population that has not been under selection pressure in the field. These results confirm the need for further studies and may guide future research with this pathogen.     

Activin A as a novel biomarker of equine inflammatory abdominal disease: preliminary findings

This study evaluated the inflammatory mediator activin A in horses with acute abdominal disease and compared this putative novel biomarker with serum amyloid A (SAA). Thirty-three adult horses referred for evaluation of acute abdominal disease were grouped into three lesion categories, non-strangulating, strangulating or inflammatory. Eleven healthy adult horses served as controls. Serum activin-A was significantly increased in animals with inflammatory or strangulating lesions compared with controls. Horses with non-strangulating, strangulating or inflammatory lesions had significantly elevated SAA concentrations. Activin A, along with other biomarkers, may be useful in monitoring inflammation in cases of acute abdominal disease in horses. Further validation is warranted to determine the utility of this biomarker in evaluating the effectiveness of novel anti-inflammatory treatments for equine colic and endotoxaemia.