Effect of the FecX(R) polymorphism in the bone morphogenetic protein 15 gene on natural or equine chorionic gonadotropin-induced ovulation rate and litter size in Rasa Aragonesa ewes and implications for on-farm application

A new mutation in the bone morphogenetic protein 15 (BMP15) gene (FecX(R) allele) causing increased prolificacy in heterozygous (R+) and sterility in homozygous ewes has been recently described in Rasa Aragonesa, a low-prolificacy Mediterranean breed. The current study determined, first, the effect of this polymorphism on natural and eCG-induced ovulation rate (OR) and the effect of eCG dose on reproductive performance; and second, its effect on prolificacy and its interaction with progestagen + eCG treatment on farms, which have not been reported to date. The FecX(R) allele increased OR by 0.44 and 0.63 ovulations in young (n = 91) and adult (n = 84) R+ ewes, respectively (both, P < 0.01), increments less than reported in prolific breeds carrying other mutations in BMP15. When the standard dose of eCG used on farms (480 IU) was applied to R+ ewes (n = 36), an extremely high OR (3.95) was recorded, which was accompanied by greater partial failure of multiple ovulations (PFMO). On the contrary, OR using 240 IU in R+ ewes (2.90; n = 35) was similar to 480 IU in wildtype (++) ewes (2.82; n = 48; both P < 0.01 when compared with 480 IU in R+ ewes). No differences were found in the birth weight of the offspring between R+ and ++ eCG-stimulated ewes within the same litter size. To validate the genealogy identification on farms, PCR genotyping was carried out in 1,667 ewes from 4 elite flocks, resulting in a negligible misclassification of R+ ewes, which demonstrated that identification by genealogy is a reliable tool to identify FecX(R) ewes within the breeding program. In recorded farms, the natural litter size of ++ ewes (1.34, n = 599,160 lambing records) was increased due to the FecX(R) allele by 0.35 lambs (P < 0.0001, n = 6,593 lambing records). A similar increase (0.30) was observed when comparing ++ and R+ ewes treated with 480 IU of eCG (P < 0.0001, n = 62,055 and n = 866, respectively). When applying 480 IU of eCG to R+ ewes, the increase in prolificacy was only due to increased percentages of triplets (P < 0.001) and quadruplets (P < 0.0001), but not of twin births. In conclusion, the favorable reproductive performance of R+ ewes, with 0.63 extra ovulations and 0.35 extra lambs per lambing ewe, is responsible for the increased interest in the use of this polymorphism. Nevertheless, care must be taken in the application of eCG to R+ ewes, with the current results showing that the standard dose increases prolificacy by only increasing triple and higher-order births.     

Characterization and pathogenicity of Pestalotiopsis uvicola causing black leaf spot on carob (Ceratonia siliqua L.) in Italy

During spring 2013, several carob plants (Ceratonia siliqua L.), showing symptoms of necrotic black lesions on leaves, were observed in Capri Island, Southern Italy. Acervuli, producing conidia with septa and appendages, appeared in the centre of the lesions. In this report, the identification of the causal agent of this disease was made applying Koch’s postulates. Morphological characteristics of colony (colour and mycelium appearance) and conidia (size, shape, number of septa, length and number of apical and basal appendages) were characterized to identify the pathogen. Moreover, for a rapid and unambiguous identification of the fungal species, the internal transcribed spacers, comprising the 5.8 rDNA gene (ITS1-5.8-ITS2), were amplified by PCR, using the genomic DNA extracted from the isolated colonies, and sequenced. The fungus isolated was ascribed to the species Pestalotiopsis uvicola (Speg.) Bissett. Phylogenetic analysis, performed with sequences present in GenBank database, showed that carob isolates clustered with P. uvicola isolates from North America and Italy, separately from Australian, Indian and Chinese isolates. P. uvicola isolates were pathogenic when artificially inoculated on carob leaves. To the best of our knowledge, this is the first report of P. uvicola on carob.     

Evaluation of a new natural adjuvant obtained from locust bean gum to reduce the amount of copper necessary to control downy mildew of grapevine

Journal of Plant Diseases and Protection - Two field trials in integrated cultivation (2014 and 2016) were carried out with the aim to evaluate the efficacy and the persistence of copper...     

The efficiency of in vitro ovine embryo production using an undefined or a defined maturation medium is determined by the source of the oocyte

In vitro oocyte maturation can be influenced by oocyte source and maturation media composition. The aim of the present study was to compare the efficiency of a defined in vitro maturation medium (TCM199 supplemented with cysteamine and epidermal growth factor; Cys + EGF) with an undefined medium (TCM199 supplemented with follicle-stimulating hormone and follicular fluid; FSH + FF) for in vitro production (IVP) of ovine embryos, using oocytes obtained by laparoscopic ovum pick-up from FSH-stimulated [n=11; 158 cumulus-oocyte complexes (COCs)] and non-stimulated (n=16; 120 COCs) live ewes, as well as abattoir-derived oocytes (170 COCs). The produced blastocysts were vitrified and some of them were transferred to synchronized recipients. The best and the worst final yields of embryo IVP observed in this study were obtained using oocytes from FSH-stimulated ewes matured in FSH + FF (41.3%; 33/80) and in Cys + EGF (19.2%; 15/78) medium, respectively (p<0.01). No significant differences between both media were attained in the blastocyst development rate or in the final yield of embryo IVP using oocytes from non-stimulated ewes or abattoir-derived oocytes. The overall in vivo survival rate of the transferred vitrified blastocysts was 13.1% (8/61), without significant differences between oocyte sources or maturation media. In conclusion, under the experimental conditions of the present study, TCM199 supplemented with cysteamine and EGF is a convenient defined maturation medium for IVP of embryos from oocytes of live non-stimulated ewes or from oocytes of abattoir-derived ovaries. However, the best final yield of embryo IVP observed in this study was attained when oocytes came from FSH-stimulated donors and TCM199 was supplemented with FSH and follicular fluid.     

Characterization and pathogenicity of Fusarium solani causing foot rot on hemp (Cannabis sativa L.) in Southern Italy

Journal of Plant Diseases and Protection - Symptoms of foot rot were observed on hemp (Cannabis sativa) plants in Campania region (Southern Italy) in 2018. The symptoms consisted of brownish areas...     

Influence of the FecXR Allele in Heterozygous Ewes on Follicular Population and Outcomes of IVP and ET using LOPU‐Derived Oocytes

Ewes heterozygous for the FecX^R allele (R+) in the bone morphogenetic protein 15 (BMP15) gene display increased ovulation rate and prolificacy. Besides this phenotypic advantage, the influence of the FecX^R allele on follicle number and size, oocyte competence and in vitro production (IVP) remains undefined. With these aims, 8 R+ and 8 wild‐type (++) ewes were subjected to 2 laparoscopic ovum pick‐up (LOPU) trials (four sessions per trial; two with and two without FSH) and subsequent IVP and fresh embryo transfer. All follicles >3 mm were punctured (n = 1673). Genotype did not significantly affect the number of punctured follicles per ewe and session (10.4 and 10.2 in R+ and ++ untreated ewes, 17.4 and 14.3 in R+ and ++ FSH‐treated ewes, respectively), but follicular diameter of R+ ewes was significantly reduced compared with ++ ewes (?0.2 mm in untreated and ?0.8 mm in FSH‐treated ewes; p < 0.01). R+ ewes showed higher recovery rate and increased numbers of total and suitable cumulus–oocyte complexes for in vitro maturation (IVM). Similar rates of day 8 blastocysts were observed in R+ (36.1%, 147/407) and ++ (32.6%, 100/307) ewes, but the final output of day 8 blastocysts per ewe and session was higher in R+ ewes (+0.75; p < 0.005), without differences in survival rate at birth of the transferred embryos (40.4%, 21/52 vs 36.4%, 16/44, respectively). In conclusion, a higher number of oocytes proven to be competent for in vitro development and embryo survival after transfer are recovered from R+ ewes, despite the lower mean size of their follicles at puncture.