Individuals naturally carry bacteria and other microbes as part of their natural flora, with some being opportunistic pathogens. Approximately 30% of the population is known to carry Staphylococcus aureus in their nasal cavity, an organism that causes infections ranging from soft tissue abscesses to toxic shock syndrome. This problem is compounded by the presence of antibiotic-resistant strains such as Methicillin-Resistant Staphylococcus aureus (MRSA). Commensal bacteria present on cadavers pose a risk to those who handle the body. As a Medical School Anatomy laboratory that performs hands-on cadaveric dissection, we wanted to know whether the embalming process is sufficient to kill all commensal bacteria that pose a risk to staff and students. Even if these strains do not cause disease in these individuals, secondary transmission could occur to friends and family, who may be at higher risk of acquiring an infection. Embalming is assumed to eliminate all microbial contamination on the body. However, there are limited studies to confirm this. This study characterises the incidence of antibiotic sensitive and resistant bacteria in cadavers donated for medical teaching and research. We have screened for Methicillin-Resistant Organisms (MRO) and Extended-Spectrum Beta-Lactamase (ESBL) producing bacteria. In this study group of cadavers, approximately 46% (16/35) carry an MRO, while 51% (18/35) carry an ESBL positive organism prior to embalming. By determining the organisms’ presence pre- and post-embalming, we can evaluate the embalming procedure’s effectiveness. Our results show embalming eliminates detectable microbes in about 51% (18/35) of the cadavers. MRO dropped by 75% (16 to 4 positive cadavers), while ESBL organisms went down by almost 95% (from 18 to 1 positive cadaver). There was a further decrease in the number of positive cadavers after storage at 4 °C to 6% (2/32). Thus, although the embalming process does not immediately sterilise all the cadavers, prolonged storage at 4 °C can further reduce the number of viable bacteria.

SPRASA (SPACA3, SLLP1) is a newly identified protein that belongs to the c-type lysozyme/alpha-lactalbumin super family. The mature SPRASA protein is 51% similar to c-type lysozyme and 34% to alpha-lactalbumin. Unlike c-type lysozyme, SPRASA does not appear to have the ability to cleave substrates but is more likely to be a substrate-modifier, similar to the function of alpha-lactalbumin. Previous investigations have shown that an antiserum generated against human SPRASA is cross reactive with sperm from red and fallow deer, elk, and bull sperm indicating that a SPRASA orthologue protein is expressed by the sperm of these animals. The aim of this study was to investigate the evolutionary conservation of the SPRASA gene sequence in orthologues from various species. Computer based analysis was used to identify known orthologues, and these were aligned against the NCBI human Sprasa (accession NP_776246) and lysozyme (accession NP_000230) genes. Degenerate primers were designed to exon 3 (human) from these orthologues and genomic DNA was extracted from either blood or tissue samples from 20 mammalian, seven avian, and one reptilian species. All samples were investigated for the presence of SPRASA gene orthologues by PCR and a number of PCR products were further analysed by direct sequencing. In silico analyses were used to convert nucleotide to amino acid sequences, align all orthologue sequences and to create a 3D image of the human SPRASA protein. This analysis indicated that SPRASA orthologues are restricted to mammalian species only. Amino acid sequence alignment of SPRASA orthologues revealed a unique motif that is not seen in other lysozyme family members. Computer modelling showed that this eight amino acid SPRASA motif is located on a surface exposed loop of the 3D image that lies within a region analogous to the substrate binding region of c-type lysosymes/alpha-lactalbumin. In conclusion, the sperm-specific protein SPRASA appears to be present only in mammals and contains a unique eight amino acid motif that is conserved across the orthologues. That SPRASA orthologues have been conserved during mammalian evolution suggests that SPRASA has an important function in reproduction. Acknowledgement: We would like to thank the Auckland and Hamilton Zoos; Dr. A. Snell, Kumeu Veterinary Service; Dr. M. Green, Liggins Institute; Dr. J. Dean, University of Auckland, for their kind donation of blood and or tissue samples. This study was supported by the Marsden Fund and The University of Auckland Staff Research Fund.

SPRASA (SPACA3, SLLP1) is a newly identified protein that encodes a single pass type II membrane protein, which is retained within the equatorial region, as well as the inner acrosomal membrane of sperm. Antibodies reactive with SPRASA have been identified in some infertile men and an antiserum reactive with recombinant SPRASA prevented human sperm binding to hamster oocytes in vitro, indicating an important role in sperm oocyte recognition. DNA sequence analysis has indicated that SPRASA is a member of the c-type lysozyme/ alpha-lactalbumin family and in silico analysis suggests SPRASA is expressed in the testes/sperm, with little or no expression in other tissues. The aim of this study was to investigate the spatial and temporal expression of SPRASA in a mouse model. Tissue samples from seven organs (brain, thymus, heart, spleen, kidney, liver and the reproductive organs) from triplicate females and males of two different strains of mice (C57 and Balb/C) were collected from duplicate animals at five postnatal time points (P1, P10, P20, P40 and P60). Total RNA was extracted, reverse transcribed and analysed by quantitative realtime RT-PCR. This analysis indicated that SPRASA mRNA was consistently expressed in the reproductive organs of both female and male mice from postnatal day 10 through to the end time point of day 60 of both strains. Low levels of SPRASA mRNA were also present in the heart of both males and female mice at each of these time points. To date the expression of SPRASA has only been reported in the testes/sperm with an additional single EST identified in the brain. Quantitative real-time RT-PCR analysis has also identified SPRASA expression in the female reproductive organs. The exact location of this expression and functional significance of this expression in the female reproductive tract is unknown. The significance of the expression of SPRASA in the heart is currently unclear, but the limited expression pattern of SPRASA does suggest a role in reproduction. Acknowledgement: We would like to thank the Marsden Fund and The University of Auckland Staff Research Fund for supporting this study.