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.     

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.     

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.     

Multidrug- and methicillin resistant Staphylococcus pseudintermedius as a cause of canine pyoderma: a case report

A case of a dog with a long-term inflammatory skin disorder due to infection with methicillin-resistant Staphylococcus pseudintermedius (MRSP) is described. After initial diagnostics of MRSP, follow-up swabs of the dog (nose, skin) were taken twice after four and seven weeks. MRSP was constantly isolated from the skin and once from the nose. Since infected humans might be a source of reinfection, the owners of the dog were screened (nasal) three times during their pet's therapy. Thereby, the male owner was found to be colonized with MRSP once in the first sampling round. Comparative typing of all MRSP-isolates by pulsed-field gel electrophoresis (PFGE), SCCmec typing, multilocus sequence typing (MLST), spa typing, PCR-detection of the leukotoxin encoding operon (LukI) and the Staphylococcus intermedius-exfoliative toxin (SIET) as well as antimicrobial resistance profiling by broth microdilution revealed that all five MRSP isolates from the dog and the single isolate from the owner were indistinguishable by any of the applied methods. All isolates were assigned to a certain strain, a multidrug-resistant MRSP belonging to sequence type (ST) 71, spa type (t)05, harbouring SCCmecIII as well as the genes encoding LukI and SIET. In this case, a number of reasons might have contributed to therapy failure and re-infection, respectively (e. g. contact to other MRSP-colonized dogs, contact to MRSP-colonized humans, refusal to clip the dog's fur). In addition, MRSP-contaminated objects or surfaces in the household, which were difficult to disinfect or simply not considered as a potential source of MRSP, might have served as a source of re-infection. These results envision the possibility of a dog-to-human transmission of MRSP and the relevance of this aspect as a potential source of re-infection in cases of bacterial-supported long-term skin disorders in canine patients. First cases of MRSP infections in humans have been described only recently. However, the general pathogenic potential of multidrug resistant MRSP in humans is unknown so far and needs further investigation.     

Haematological and histological findings in birds experimentally infected with highly pathogenic H5N2 avian influenza virus

Experimental infection of specific-pathogen-free (SPF) Leghorn chickens with a highly pathogenic H5N2 avian influenza virus produced cellular hyperplasia in the bone marrow at 36 hours post infection (hpi) and haematological evidence of monocytosis, thrombocytopenia and heterophilia was also detected. An early, significant and progressive haematological change was thrombocytopenia starting at 24 hpi without an increase of prothrombin time. The findings suggest that highly pathogenic avian influenza virus interferes only with the primary haemostatic mechanisms by consumption of thrombocytes, while the secondary haemostatic mechanisms remain intact.     

Haematological and histological findings in Leghorn chickens infected with infectious bursal disease virus strain 73688

In the present study, specific-pathogen-free, 2-week-old Leghorn chickens were experimentally infected with strain 73688 of infectious bursal disease virus (IBDV) in order to evaluate haematological and histological changes that might suggest a pathomechanism for haemorrhages in this disease. At 96 hours post infection (hpi) a significant increase in prothrombin time was detected in the absence of visible lesions in myeloid bone marrow tissue and of significant thrombocytopenia. The aforementioned findings suggest alteration of the secondary coagulation mechanisms and not a direct effect of virus on thrombocytes or its precursors.     

Thrombocytopenia in Newcastle disease: haematological evaluation and histological study of bone marrow

A Newcastle disease virus (NDV) isolated in Mexico and called Chimalhuacan strain was characterised by gene F restriction enzyme analysis and found to be a genotype II velogenic virus. Haematological evaluations and histological studies of bone marrow were conducted on chickens experimentally infected with the Chimalhuacan virus and on control chickens. Within 72 hours post infection (hpi), a 50% decrease in thrombocyte and monocyte counts and a complete cellular depletion in bone marrow islands were evident in the infected group. These findings suggest that the Chimalhuacan strain of NDV causes an early and severe damage of the haematopoietic cells including thrombocyte precursors, which might explain the marked thrombocytopenia detected in early stages of this disease.     

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.