Production of Lambs After the Transfer of Fresh and Cryopreserved in vitro Produced Embryos from Prepubertal Lamb Oocytes and Unsorted and Sex-sorted Frozen-thawed Spermatozoa

Cumulus-oocyte complexes from hormone-stimulated 3-4-week-old (n=43) and 6-7-week-old (n=12) prepubertal lambs were matured in vitro and incubated with unsorted, or X- or Y-spermatozoa separated with a high-speed cell sorter (SX MoFlo)frozen-thawed. Presumptive zygotes were then cultured to the blastocyst stage, and transferred to recipients fresh or after cryopreservation (frozen). Oocyte cleavage was higher (p <0.05) with unsorted (515/926, 55.6%) than X- or Y-spermatozoa (261/672, 38.8% and 229/651, 35.2%, respectively) and blastocyst formation (% zygotes) by Day 9 of in vitro culture was lower (p <0.05) for X- (102/261, 39.1%) than unsorted spermatozoa (249/515, 48.3%), but did not differ between Y-spermatozoa (103/229, 45.0%) and unsorted spermatozoa, or between X- and Y-spermatozoa (p >0.05). For fresh embryos, survival to term was 50.0% (3/6) for unsorted, 0.0% (0/6) for X- and 16.7% (1/6) for Y-spermatozoa-derived embryos (p >0.05), and for frozen embryos was 4.0% (2/50) for unsorted, 9.1% (2/22) for X- and 2.9% (1/34) Y-spermatozoa-derived embryos (p >0.05). Of the two lambs born from X-spermatozoa-derived embryos, one was female (50%), and from the two Y-spermatozoa-derived lambs, both were male (100%), demonstrating that lambs can be produced after the transfer of fresh and cryopreserved IVP embryos derived from prepubertal lamb oocytes and frozen-thawed sex-sorted sperm.     

Molecular characterization of the incitant of cowpea bacterial blight and pustule, Xanthomonas campestris pv. vignicola

Strains of Xanthomonas campestris pv. vignicola (Xcv), isolated from cowpea leaves with blight or minute pustules and collected from various geographic areas, were selected on the basis of pathological and physiological features. All strains were analyzed for genotypic markers by two methods: ribotyping with EcoRI endonuclease, and RFLP analysis with a plasmid probe (pthB) containing a gene required for pathogenicity from Xanthomonas campestris pv. manihotis. Ribotyping revealed a unique pattern for all the strains that corresponded to the previously described ribotype rRNA7. Based on polymorphism detected by pthB among Xcv strains, nine haplotypes were defined. The observed genetic variation was independent of the geographic origin of the strains and of pathogenic variation. Some haplotypes were widely distributed, whereas others were localized. In some cases, we could differentiate strains isolated from blight symptoms and pustules according to haplotypic composition. However, in most cases, no significant differences were observed. Our results and the previous pathogenic and biochemical characterizations suggest that the strains isolated from leaves with blight symptoms or minute pustules belong to the same pathovar. We provide information on pathogen diversity that can be used to identify and characterize resistant germplasm.     

Device Applications of Side-Chain Ferroelectric Liquid Crystalline Polymer Films

Side-chain ferroelectric liquid crystalline polymers are currently used in a number of applications, including displays and electrical sensors. Comparisons with existing technologies and materials indicate that relative to ceramics, such polymers have lower figures of merit but offer greater durability in sensor applications.     

Minerals content of extruded fish feeds containing cricket ( Acheta domesticus ) and black soldier fly larvae ( Hermetia illucens ) fractions

Animal food sources provide human beings with minerals considerably in adequate quantities. Fish is an indispensable reliable source of nutrients, as aquaculture is a sector that is fast growing and which provides 50% of the world’s fish production. However, fish production is hampered by the increasing costs of feeds due to the ever rising cost of fish meal, an integral component of fish feeds. Substituting fish meal with cheap, yet highly nutritious ingredients in fish feeds is therefore paramount. This study investigated the effects of substituting fish meal with adult cricket meal (ACM) and black soldier fly meal (BSFM) on minerals content of extruded fish feeds, where four levels of substitution (0, 25, 50 and 75%) were used. The effect of feed moisture content on minerals was also studied where 20 and 30% feed moisture levels were used. Leaching effects of the pellets were studied as well. The results showed a significant increase (P < 0.05) in the levels of phosphorus and potassium as the level of fish meal substitution increased from 0 to 75%. On the other hand, iron and sodium levels reduced significantly (P < 0.05) as the level of fish meal substitution increased. Magnesium content increased with increasing level of substitution with BSFM, but decreased with increasing level of substitution with ACM. Copper, zinc and manganese were not greatly influenced by levels of fish meal substitution. Diets that had zero substitution showed higher leaching effect for most minerals than diets that were substituted with 75% ACM or BSFM. This study found that both ACM and BSFM can be used to substitute fish meal in fish feeds and obtain adequate mineral profile and low leaching effect.     

Incomplete systemic movement of Xanthomonas campestris pv. musacearum and the occurrence of latent infections in xanthomonas wilt‐infected banana mats

Management of banana xanthomonas wilt (XW) (caused by Xanthomonas campestris pv. musacearum, Xcm) has been impeded by poor adoption of control options that are complex, cumbersome and costly. To improve XW management, this study investigated Xcm survival and latent infections in subsequent generations, survival of latently infected planting materials (suckers), incidence of latent infections in symptomless plants in mats having diseased plants, and XW status across farms and markets in districts previously devastated but currently endemic. On‐station experiments were protected from new infections. Latent bacteria at low levels were detected in up to 20% of the third generation suckers, with a significant (P < 0·05) reduction (43–20%) in subsequent generations. Only 3–6% of latently infected suckers succumbed to XW. Incidence of Xcm in symptomless suckers from farmers' fields (with up to 70% incidence) was low (3%) while it increased (8–25%) with disease severity in mats in controlled experiments. In the surveyed districts, incidence had significantly declined with yields observed to have recovered relative to earlier reports, although latent infections remained high. This study provides evidence that if new infections are prevented, fields with high XW incidence can be rejuvenated. It showed incomplete systemic movement of Xcm in mats coupled to a gradual decline of bacterial load in subsequent generations to levels that cannot initiate disease. These studies explain the current successes in farms practising single diseased plant removal instead of whole mat rouging, and gives hope to farmers lacking access to clean planting material.     

Impact of Distillery Effluent on Soil Carbon and Nitrogen Dynamics in a Vertisol of Central India

Distilleries produce a huge quantity of effluents, popularly known as spent wash (SW), which when bio-methanated produce post-methanation effluents (PME). A field experiment on soybean–wheat system was conducted for five consecutive years in a Vertisol of central India to evaluate the effect of distillery effluent (DE) on soil carbon and nitrogen dynamics. Ten treatment combinations consisting of control, 100% NPK + Farmyard Manure (FYM), and graded level of SW and PME were applied. Total carbon content of soil increased significantly with applications of FYM and DE. SW was found superior in enhancing carbon content of soil in comparison to PME. Farmyard Manure contributed more carbon toward the recalcitrant pool, whereas DE contributed more carbon toward the active and slow pool. Nitrogen (N) availability was significantly improved with the application of DE. Balanced application of DE may act as amendment for increasing C and N stocks in Vertisol.     

The acquisition of cadmium by Lupinus albus L., Lupinus angustifolius L., and Lolium multiflorum Lam.

Das Cadmiumaneignungsvermögen von Lupinus albus L., Lupinus angustifolius L. und Lolium multiflorum Lam. Mehrere Pflanzenarten mobilisieren Bodenphosphate (P) und Kationen wie Fe und Al durch die Exsudation organischer Anionen und Protonen. Deshalb untersuchten wir das Cd‐Aneignungsver‐mögen von P‐, Fe‐, Al‐mobilisierenden Arten (Lupinus albus L., Lupinus angustifolius L.) im Vergleich zu einer nicht mobili‐sierenden Pflanzenart (Lolium multiflorum Lam.). Die Pflanzen wuchsen in zwei stark unterschiedlichen Böden (saurer Humuspodsol, karbonathaltiger Lössunterboden). Die Cd‐Aufnahme in die Sprosse war bei Weidelgras 5 bis 10 mal höher als bei Blauen bzw. Weißen Lupinen. Dieses Ergebnis hat mehrere Ursachen: 1. Das Wurzellängen/Sprossmasseverhältnis des Weidelgrases ist 2—3 mal größer als das der Lupinenpflanzen. 2. Bei Weidelgras wird ein größerer Teil des aufgenommenen Cd in die Sprosse verlagert. 3. Die Cd‐Aufnahme bei Lupinen ist im sauren Boden (Podsol) und bei P‐Mangel auch im Kalkboden niedriger als bei Weidelgras. Während im Podsol die Cd‐Konzentration der Bodenlösung unter Lupine geringer war als in der Kontrolle (Gefäße ohne Pflanzen), war sie im Kalkboden höher. Bei den Lupinen war der Efflux organischer Säureanionen, vor allem Citrat und Malat, um den Faktor 10—100 höher als bei Weidelgras. Diese Exsudation kann zu einer hohen Cd‐Komplexierung, insbesondere durch Citrat, in der Rhizosphärenbodenlösung führen (˜ 85%). Diese Ergebenisse deuten darauf hin, dass das komplexierte Cd von den Wurzeln schlechter aufgenommen wird als das freie Cd.     

Isolation and molecular characterisation of Neospora caninum in cattle in New Zealand

AIM: To isolate Neospora caninum from the brains of naturally infected cattle and use molecular techniques to characterise the isolates.

METHODS: Neospora caninum tachyzoites were isolated in Vero cell culture from the brains of a cow and two calves. The isolates were characterised using polymerase chain reaction (PCR) methods, DNA sequencing, an immunofluorescent anti-body test (IFAT), transmission electron microscopy (TEM), and immunohistochemistry (IHC). The brains of the three cattle were subjected to histopathological examination. A pathogenicity study was conducted in 120 BALB/c mice.

RESULTS: Neospora caninum tachyzoites were isolated from all three cases and first observed in vitro between 14 and 17 days post-inoculation. Parasites were sub-cultured and maintained in Vero cell culture for more than 6 months. PCR products were generated for all three isolates, using two different primers. Sequencing of the PCR products and a subsequent BLAST search identified the isolates as N. caninum. In addition, the isolates tested positive using IFAT and IHC, and ultrastructure revealed by TEM was characteristic of N. caninum. Histopathological examination revealed lesions characteristic of N. caninum in 1/3 brains. In the pathogenicity study using BALB/c mice, the mortality rate was 3–7%.

CONCLUSION: This was the first successful isolation of N. caninum in New Zealand confirmed using molecular characterisation tests.