Effect of Treatment with Heated Scallop Shell Powder on the Inactivation of Naturally Existing Bacteria and Listeria monocytogenes Inoculated on Chicken Meat.

This study investigated the efficacy of heated scallop shell powder (HSSP) treatment in preserving chicken thigh meat. Chicken thigh meat was treated with HSSP slurry (1% and 5%) for 60 min, and the variation in aerobic bacteria and coliform populations was assessed during refrigerated storage (10 °C). There was a substantial increase in aerobic bacteria, reaching nearly 7 log10 colony forming unit (CFU)/g following 7 days of refrigeration, in the untreated chicken meat. Conversely, the aerobic bacterial population of the HSSP-treated chicken was <5 log10 CFU/g. Coliform growth in the untreated chicken reached over 5 log10 CFU/g following 7 days. In contrast, the coliform population of the HSSP-treated chicken did not reach 5 log10 CFU/g at 1% HSSP concentration; it was suppressed to <4 log10 CFU/g at 5% concentration. Listeria monocytogenes, which can grow at low temperatures, was inoculated into the chicken meat (5 log10 CFU/g) treated with alcohol, which was followed by HSSP. In the untreated chicken, L. monocytogenes increased to 9 log10 CFU/g even when refrigerated for 7 days. However, in the chicken treated with 5% HSSP, L. monocytogenes was suppressed to approximately 3 log10 CFU/g. These findings reveal that HSSP treatment is an effective method for disinfecting meat, inhibiting bacterial growth, and enhancing preservation.

Relation between semen oxidative reduction potential in initial semen examination and IVF outcomes.

Purpose: The MiOXSYS system is a new technique to analyze the semen oxidative reduction potential (ORP) that may use to classify the level of sperm DNA integrity. It does not clearly explain how the semen ORP values could help to change the IVF outcomes. We have analyzed correlations between semen ORP value and the IVF results. Methods: Four hundred and thirty couples were enrolled. The male counterparts were divided into two groups according to their semen ORP values and compared the fertilization rate, cell cleavage rate, and embryo quality, following the IVF procedures. The relations between ORP values and the clinical pregnancy, live birth, and abortion rates were analyzed. Results: The ORP values show negative and positive correlations with some conventional semen parameters. The fertilization and the cleavage rate did not show any differences in those two groups, but the transferable embryo rate was significantly high in patients with high semen ORP. However, the patients with high ORP show a tendency to lower clinical pregnancy with a low abortion rate compared to the low ORP group. Conclusion: The main purpose of measuring the ORP value in semen is still questionable and shows controversial results.

Male Oxidative Stress Infertility (MOSI): Proposed Terminology and Clinical Practice Guidelines for Management of Idiopathic Male Infertility.

Despite advances in the field of male reproductive health, idiopathic male infertility, in which a man has altered semen characteristics without an identifiable cause and there is no female factor infertility, remains a challenging condition to diagnose and manage. Increasing evidence suggests that oxidative stress (OS) plays an independent role in the etiology of male infertility, with 30% to 80% of infertile men having elevated seminal reactive oxygen species levels. OS can negatively affect fertility via a number of pathways, including interference with capacitation and possible damage to sperm membrane and DNA, which may impair the sperm's potential to fertilize an egg and develop into a healthy embryo. Adequate evaluation of male reproductive potential should therefore include an assessment of sperm OS. We propose the term Male Oxidative Stress Infertility, or MOSI, as a novel descriptor for infertile men with abnormal semen characteristics and OS, including many patients who were previously classified as having idiopathic male infertility. Oxidation-reduction potential (ORP) can be a useful clinical biomarker for the classification of MOSI, as it takes into account the levels of both oxidants and reductants (antioxidants). Current treatment protocols for OS, including the use of antioxidants, are not evidence-based and have the potential for complications and increased healthcare-related expenditures. Utilizing an easy, reproducible, and cost-effective test to measure ORP may provide a more targeted, reliable approach for administering antioxidant therapy while minimizing the risk of antioxidant overdose. With the increasing awareness and understanding of MOSI as a distinct male infertility diagnosis, future research endeavors can facilitate the development of evidence-based treatments that target its underlying cause.

Characteristics of Biofilms Formed by Co-Culture of Listeria monocytogenes with Pseudomonas aeruginosa at Low Temperatures and Their Sensitivity to Antibacterial Substances.

We assessed the properties of biofilms (BFs) formed by mono- and co-cultures of Listeria monocytogenes and Pseudomonas aeruginosa (L+P-BF) at low temperatures and examined their sensitivity to several antibacterial substances. L. monocytogenes viable counts comprised only 1-10% of total L+P-BF viable counts at 10℃ and 15℃, indicating the significant prevalence of P. aeruginosa in co-cultures. L+P-BF formed at 10℃ and 15℃ showed very high resistance to antibiotics and NaClO. Examination of the effects of nattokinase and nisin, natural food additives with antibacterial properties, showed that their application alone failed to inhibit L+P-BF development at 10℃ and 15℃. However, a combined treatment with nisin and ethylenediaminetetraacetic acid, a food additive that can be used as a permeabilizing agent, suppressed the formation of L+P-BF at 10℃ and 15℃. Microscopy observations of L+P-BF did not reveal pronounced morphological changes in bacterial cell morphology. We also noted that P. aeruginosa resistance to the action of nisin during BF formation was higher when it was maintained in co-culture with L. monocytogenes. The results of the present study are an important step toward developing a safe formulation of acceptable food additives that could be used for suppression of BFs formed by pathogenic bacteria during food storage.

Preincubation of Escherichia coli ATCC 25922 with NaCl Increases Its Attachment to Lettuce Surfaces Compared with Other Chemicals.

　The inhibition of microbial attachment to food is important for the prevention of cross-contamination during food processing. The effect of several chemicals that were added in an Escherichia coli growth medium on the attachment of the bacterium to lettuce was investigated. E. coli ATCC 25922, which is reportedly a useful surrogate for E. coli O157:H7 in surface attachment studies, was preincubated in a nutrient broth (NB) containing sodium chloride, potassium chloride, sodium deoxycholate, sodium linear alkylbenzene sulfonate, or sorbic acid. The bacterial cells were placed in contact with cut lettuce in a saline solution at 5℃ for 24 hours. Only the addition of NaCl in the NB influenced the attachment of E. coli, Salmonella enterica subsp. Enteritidis, and Klebsiella pneumoniae to the lettuce. The attachment of E. coli showed the largest significant increase at 2% NaCl. Changes in the attachment levels were not due to surface hydrohobicity or the motility of E. coli cells. Similar results were observed for S. enterica although the variation in the degree of attachment of the latter was quite small. These results suggested that the attachment of E. coli O157:H7 to food surfaces is influenced by the bacterial growth conditions prior to food exposure and prior to the development of the biofilm; furthermore, the environmental NaCl concentration should be controlled during food processing to prevent the cross-contamination of foods with E. coli.

Combination of density gradient centrifugation and swim-up methods effectively decreases morphologically abnormal sperms.

Density gradient centrifugation (DGC) and swim-up techniques have been reported for semen preparation in assisted reproductive techniques in humans. We investigated whether semen preparation using a combination of DGC and swim-up techniques could effectively decrease morphologically abnormal human sperms at the ultrastructural level. Semen samples were obtained from 16 infertile males and fractionated by swim-up following DGC. Ultrastructural abnormalities of sperms obtained from original semen, lower layer of swim-up following DGC, and upper layer of swim-up following DGC were analyzed by transmission electron microscopy. The correlation among ultrastructural head abnormality in sperms from the upper layer of swim-up, fertilization in in vitro fertilization, and pregnancy after embryo transfer was also investigated. Furthermore, sperms with DNA fragmentation in the samples processed via a combination of DGC and swim-up was assessed in a sperm chromatin structure assay. Ultrastructural abnormalities in sperm heads and tails in the upper layer after swim-up following DGC was the lowest among the three groups. Sperms with nuclear vacuoles were the most difficult to eliminate using a combination of DGC and swim-up in all types of head abnormalities. A negative correlation was confirmed between the fertilization rates of intracytoplasmic sperm injection and head abnormality of sperms obtained from the upper layer of the swim-up following DGC. Sperms with DNA fragmentation were effectively decreased using the combination of two techniques. In conclusion, the combination of DGC and swim-up effectively decreased the number of sperms with ultrastructural abnormalities both in the head and in the tail. However, sperms with ultrastructural abnormalities that cannot be completely decreased using a combination of DGC and swim-up may impair fertilization in some cases of intracytoplasmic sperm injection.

Cryopreservation of a small number of human sperm using enzymatically fabricated, hollow hyaluronan microcapsules handled by conventional ICSI procedures.

PURPOSE: We investigated whether enzymatically fabricated hyaluronan (HA) microcapsules were feasible for use in the cryopreservation of a small number of sperm. METHODS: HA microcapsules were fabricated using a system of water-immiscible fluid under laminar flow. Three sperm were injected into a hollow HA microcapsule using a micromanipulator. Capsules containing injected sperm were incubated in a freezing medium composed of sucrose as the cryoprotectant and then placed in a Cryotop® device and plunged into liquid nitrogen. After thawing, the capsule was degraded by hyaluronidase, and the recovery rate of sperm and their motility were investigated. RESULTS: The HA microcapsule measuring 200 µm in diameter and with a 30-µm thick membrane was handled using a conventional intracytoplasmic sperm injection (ICSI) system, and the procedure involved the injection of sperm into the capsule. The HA microcapsules containing sperm were cryopreserved in a Cryotop® device and decomposed by the addition of hyaluronidase. The recovery rate of sperm after cryopreservation and degradation of HA microcapsules was sufficient for use in clinical practice (90 %). CONCLUSIONS: Hollow HA microcapsules can be used for the cryopreservation of a small number of sperm without producing adverse effects on sperm quality.