The objective of this study is to establish the shelf life of non-pasteurized whole egg, egg yolk and egg white liquid. Each sample was stored for two weeks at 5oC, 10oC, 15oC, and 25oC, and then sensory, microbial, and physicochemical tests were performed periodically. The estimation of shelf life was based on the microbial standards of total viable counts and coliforms. The chemical properties highly correlated with the sensory evaluation were also used. Our results showed that the shelf life was the most influenced by microbial properties. Exceptionally, however, whole egg and white liquid stored at 5oC and 10oC with limited bacterial growth were affected by chemical property. The shelf life of the three non-pasteurized liquids was calculated to be less than one day at over 15oC. At 5oC and 10oC, the shelf life was calculated to be 5 d and 1 d for egg yolk liquid, 5 d and 5 d for egg white, and 7 d and 5 d for whole egg, respectively. Therefore, it is advisable to establish reasonable shelf life in the more specific manner based on consideration of these findings.

본 연구에서는 국내 대표 식육인 소, 돼지, 닭, 오리의4종 식육과 염소, 양, 말, 칠면조의 4종 식육을 동시에 신속하게 감별할 수 있는 2 set의 multiplex PCR법을 개발하고자 미토콘드리아 16S RNA에서 종 특이부위를 선발하고 각 종에 대한 특이도를 높이기 위하여 인위적인 미스매치를 주어 프라이머를 제작한 후 8종 식육의 274개시료를 대상으로 특이도와 민감도를 조사하였다. 그 결과소, 돼지, 닭, 오리 모든 시료에서 각각 279, 94, 192, 477 bp의 증폭산물이, 말, 양, 염소, 칠면조의 모든 시료에서 각각 152 bp, 271 bp, 670 bp, 469 bp에서 뚜렷한 PCR 유전자 산물이 확인되어 모든 축종에서 100%의 특이도를 나타내어 축종별 감별력이 우수한 것으로 나타났다. 8종의 축종별로 DNA를 10 ng/μl으로 정량한 후 혼합물을 10배씩 단계 희석하여 반응여부를 조사한 결과, 소, 돼지, 오리에서는 100 fg까지, 닭에서는 1 pg까지 검출됨을 확인할수 있었다. 소, 돼지, 닭, 오리고기를 99.9%, 99%, 90%,70%, 50%, 30%, 10%, 1%, 0.1%의 비율로 혼합한 식육과 83℃ 20분, 100℃ 30분, 121℃ 10분에서 각각 열처리한 가열 혼합육에 대하여 검출한계를 조사한 결과 마지막단계의 희석 비율인 모든 혼합육의 0.1%에서 검출이 가능하였으며, 열처리 혼합육에서는 닭에서는 1% 농도에서소와 돼지의 혼합육에서 0.1% 농도에서 검출되어 민감도가 높음을 확인할 수 있었다. 본 연구에서 개발된 multiplex PCR법은 특이도 및 민감도에 있어서 국내 대표 식육을 감별하는데 있어서 유용한 것으로 평가된다.

본 연구는 계란가공전문업체에서 생산하는 즉석섭취 알가공품의 미생물학적 안전성을 분석한 연구로 제품의 종류에 따라 일반세균 및 coliforms의 오염수준의 차이가 매우 크며 식중독균의 증식/생존 양상에도 영향을 미치는 것으로 나타났다. 즉석섭취 알 가공품 중 계란구이의 가공 공정단계에서 미생물 오염도를 조사한 결과에 따르면 공정 초기단계에서 aerobic plate counts 오염 수준은 높았으나 가열 성형 공정 이후 급격하게 감소하여 공정 과정 이후 낮은 수준으로 유지하는 것으로 보아 HACCP 공정에서의 CCP 관리는 적절하게 수행되고 있는 것으로 나타났다. 완제품의 경우 치즈, 참치, 피자오믈렛과 계란구이, 계란 찜은 미생물 규격 기준에 부합하였으나 떡갈비오믈렛과 지단채는 허용 불가능한 수준으로 확인되었다. 특히 지단채의 경우 레토르트 공정을 거치지 않는 제조 공정에 따른 특성 때문인 것으로 판단되어 이를 보완할 수 있는 관리 방안이 필요할 것으로 보이며, 오믈렛의 경우 계란 이외의 내용물이 추가 주입되므로 부 재료의 관리도 중요한 것으로 나타났다. 또한 병원성 식중독균을 인위적으로 오염시킨 즉석섭취 알 가공품을 4, 10, 15℃에 저장하면서 미생물의 증식 및 생존을 관찰한 결과, L. monocytogenes는 저장 기간 내 모든 온도에서 증식하였다. 특히 계란구이에서 S. Typhimurium과 E. coli는 4℃와 10℃ 저장조건에서 사멸하였으나, 15℃ 저장조건에서는 동일한 저장기간 동안에 급격하게 성장하는 것으로 나타나, 유통/판매 조건에서 온도 관리 또한 철저하게 수행되어야 할 것으로 판단된다. 또한 계란구이에서 S. Typhimurium에 비해 S. Enteritidis의 증식 속도가 더 빠른 것으로 나타났으며 계란 찜에서도 S. Typhimurium는 사멸하는 반면 S. Enteritidis 증식은 잘 이루어 져 알 가공품에서의 S. Enteritidis의 증식 위험성이 더 큰 것으로 나타나 식용란이 생산되는 과 정에서도 S. Enteritidis 오염예방이 매우 중요한 것으로 판단된다. 또한 알 가공품의 경우 제조 시설에서의 위생적인 제품 생산뿐만 아니라 조리 시 적절한 가열, 가열 후 교차오염 방지, 조기섭취 등 유통/보관 후 소비자 섭취시점까지의 안전관리 방안이 필요하다.

본 연구는 축산식품과 식중독 세균 조합의 위험순위 결정을 위한 방법으로 축산식품 관련 식중독사고 보고서(2008-2012년 자료), 축산식품 전문가의 견해, 반 정량적 위해평가 도구인 Risk Ranger 를 이용하여 문헌자료를 분석 그 결과를 비교하였다. 본 연구결과에 따르면 지난 2008년-2012년 기간 동안 국내 축산식품에서 식중독 발생 빈도가 높았던 주요 원인균은 Salmonella, Pathogenic E. coli, C. jejuni 순으로 나타났으며 Salmonella 식중독의 주요 원인 식품은 계란인 것으로 보고되었다. 그러나 국내에서 우선적으로 관리가 필요하다고 생각되는 식중독 세균/축산식품조합에 대해 축산식품 전문가들은 가장 우선적 위해 관리가 필요한 Top 5순위 중 첫 번째로 Campylobacter/계육을 제시하였고 그 다음으로 Salmonella/식용란 및 알가공품, Enterobacter sakazakii/조제분유, Pathogenic E. coli/분쇄가공육, Pathogenic E. coli/식육을 선정하였으며 Salmonella 는 식육보다는 식용란 및 알 가공품에서 우선적 위해관리가 필요함이 제시되었다. 또한 분쇄가공육, 소시지 및 햄류에 대해서는 Cl. perfringens, L. monocytogenes, S. aureus 의 3가지 병원성 미생물들의 위해관리의 필요성이 제시되었다. 또한 국외에서 개발된 반 정량적 미생물 위해평가 도구인 Risk Ranger 를 이용하여 국내 식중독발생 통계자료 및 전문가 의견 설문조사 등과 비교 분석한 결과, Risk Ranger 는 식품과 미생물 조합의 위험 순위를 간단하게 평가할 수 있는 장점이 있으나 Risk Ranger 의 결과에 영향을 주는 입력변수에 필요한 데이터가 없을 경우에 최종결과를 신뢰하기에 한계를 보였다. 특히 국내에서는 축산식품 원재료에 대해 식중독 세균의 정량적 오염수준 및 공정에 따라 오염수준에 미치는 효과에 대한 자료가 매우 부족한 것으로 나타났는데 Risk Ranger 결과, 위해순위가 0으로 나타난 경우를 분석해 보면 모니터링 자료가 없거나, 섭취 전의 조리를 통해 99% 가열이 가능한 경우였다. 최종적으로 본 연구에서는 축산식품관련 식중독 발생 통계자료, 전문가의견, Risk Ranger 분석결과를 종합적으로 분석하여 식중독 세균/축산식품 조합에 대한 위해평가 및 관리 우선대상을 순위별로 그룹화하였다. 식중독 사고 발생 및 위해 가능성이 가장 높은 위험그룹 I 에는 Salmonella spp./식용란 및 알 가공품, Campylobacter spp./계육, Pathogenic E. coli/식육 및 분쇄가공육이 선정되었으므로 차후 이들 제품에 대한 정량적 위해평가 연구가 필요할 것으로 사료된다. 본 연구결과는 차후 축산식품 및 그 가공품에 대한 정량적 미생물 위해평가의 효율성을 높이고 우선 관리 대상에 대한 과학적 근거 자료를 제시할 수 있을 것으로 사료된다.

In this study, two commercial PCR and ELISA test kits were examined for identification of eight animal species (beef, pork, chicken, duck, turkey, goat, lamb, and horse) from raw meat and meat products in Korea. The detection limit in RAW meat ELISA kit® on three types of meat samples blended with beef, pork and chicken, demonstrated that all meat species were differentiable down to 0.2%. RAW meat ELISA kit® on animal species resulted in differentiation rate of 94.5% for beef, 93.3% for pork, 90% for lamb, and 100% for chicken, duck, turkey, goat, and horse. In contrast, Powercheck Animal Species ID PCR kitTM resulted in 100% specificity at 0.05% limit of detection for all meat species. The detection limit of Cooked Meat ELISA kit® on mixed meat samples heat-treated with different temperatures and times, resulted in 0.1% for all heat-treated mixed meat except for chicken at 1.0%. Additionally, ELISA kit on sixty meat products resulted in specificity of 31.8% for ham, 13.6% for sausages, and 12.5% for ground processed products, and relatively low rate for more than 2 types of mixed meats. On the contrary, meat species differentiation using PCR kit showed higher percentage than that using ELISA kit®: 50.0% for ham, 41.7% for sausages, and 28.6% for ground processed meat. Futhermore, PCR kit on 54 dried beef meats detected pork genes in 13 products whereas ELISA kit showed negative results for all products. Hence, the possibility of cross-contamination during manufacturing process was investigated, and it was found that identical tumblers, straining trays, cutters and dryers were used in both beef and pork jerky production line, suggesting the inclusion of pork genes in beef products due to cross-contamination. In this study, PCR and ELISA test kits were found to be excellent methods for meat species differentiation in raw meat and heat-processed mixed meat. However, lower differentiation rate demonstrated in case of meat processed products raised the possibility of inclusion of other species due to cross-contamination during manufacturing process.

We compared between an automated most-probable-number technique TEMPO®TVC and traditional plating methods PetrifilmTM for estimating populations of total aerobic bacteria in various livestock products. 257 samples randomly selected in local retail stores and 87 samples inoculated with E. coli ATCC 25922, Staphylococcus aureus ATCC 12868 were tested in this study. The degree of agreement was estimated according to the CCFRA (Campden and Chorleywood Food Research Association Group) Guideline 29 and the agreement indicates the difference of two kinds methods is lower than 1 log base 10(log10). The samples of hams, jerky products, ground meat products, milks, ice creams, infant formulas, and egg heat formed products were showed above 95% in the agreement of methods. In contrast, proportion of agreement on meat extract products, cheeses and sausages were 93.1%, 92.1%, 89.1%, respectively. One press ham and five sausages containing spice and seasoning, two pork cutlets containing spice and bread crumbs, two meat extract product and two natural cheeses and one processing cheese with a high fat content, and one ice cream containing chocolate of all samples showed the discrepancy. Our result suggest that TEMPO®TVC system is efficient to analyses total aerobic bacteria to compare manual method in time-consuming and laborious process except livestock products having limit of detection.

We investigated the prevalence of the Listeria monocytogenes from livestock processed products in processing plants and retail markets of Korea from 2010 to 2011. A total of 1,380 samples were collected; Meat processed products such as cooked ham and sausage, jerked meat, and meat extract products. Milk processed products such as milk, butter, cheese, and ice cream. Egg processed products such as whole egg liquid and pidan. L. monocytogenes were isolated from samples using listeria enrichment broth, fraser broth and Oxford agar, and counted in Oxford agar. The three of L. monocytogenes strains (1.16%) were isolated from sausages, two (0.73%) from mixed pressed ham and one (0.51%) from jerked meat, respectively. The colony forming unit (CFU) of L. monocytogenes from all samples were below 10 CFU/g. The four isolates (66.6%) were 1/2b except two isolates (1/2a) in the serotypes. Further studies are needed to understand the transmission route of L. monocytogenes, including a survey of food handlers, environments of retail markets, and all potential risk factors in cooked sausages, mixed pressed hams, and jerked meat processing.

Horse population has been gradually increasing with the growth of the racing industry in Republic of Korea (ROK). Approximately 22,941 horses including native ponies were raised on 1,142 farms in 2006. Great care has been taken to prevent viral, bacterial and parasitic infections in horses. However, there has been few reports on equine infectious anemia (EIA) in ROK until recently since 1987. Therefore, we conducted a serological survey of EIA in ROK from 2007 through 2009. Using the agar gel immunodiffusion test, a total of 2,601 horses were tested in Foreign Animal Disease Division of Animal, Plant, and Fisheries Quarantine and Inspection Agency. This survey found no serological evidence of EIA presence in ROK. Although our surveillance found no evidence of EIA in ROK during the surveillance period, it is possible that EIA could be introduced into ROK in the near future. Increased cooperation between the government and other agencies, such as horse-racing authorities, is important for both the early detection of the disease and the development of effective veterinary and public health strategies.

To evaluate the performance of a new automated coliform enumeration system (TEMPO® CC) for the quantitative test of coliform bacteria contaminated in domestic livestock processed foods, a total of 507 samples of livestock foods were tested by the TEMPO® CC method, the most probable number (MPN) method, and Petrifilm method, respectively. The results of those three methods were compared to each other. Of 507 samples of livestock processed foods used in this study, 217 samples were contaminated artificially with coliform bacteria and the rest (n=290) were contaminated naturally. The results of the TEMPO® CC method for all samples were equivalent to those obtained from the MPN method, except 8 samples. In addition, 496 (97.8%) out of 507 samples made agreement between the TEMPO® CC method and the Petrifilm method. The correlation coefficients between TEMPO® CC and MPN methods as well as between TEMPO® CC method and Petrifilm method were above 0.9, and the slope and intercept of the linear regression model was different in less than 1 value. In conclusion, there were statistically equivalent levels of performance between the TEMPO® CC and the reference and alternative methods for the enumeration of coliform bacteria in livestock processed foods in this study.

Three-hundred samples of powdered infant formula milk and related products from four different manufacturers in 2010 were collected and surveyed their contaminations for aerobic bacteria, coliform, Enterobacter (Cronobacter) sakazakii, and food-borne pathogens. Fifteen samples of sterilized infant formula milk were all negative on these microorganisms. In all collected products of unsterilized infant formulas and follow-on infant formulas,aerobic bacteria were detected at 239 (83.9%) among 285 samples, and they all were found below 10³ cfu/g. Coliform bacteria were also detected at four among 285 samples. Salmonella spp. and Ent. sakazakii, weren't detected at the all samples. Bacillus cereus was detected at 24 (8.4%) among 285 samples. The level of B. cereus was below 100 cfu/g but it was suitable for the range of specification of B. cereus in infant formulas. Clostridium perfringens, Escherichia coli O157:H7, Staphylococcus aureus and Listeria monocytogenes weren't also detected. In consequence, it was suitable for total viable count, coliform and potential pathogen to the specification of infant formulas and related products.

We constructed a standard curve to quantify Listeria monocytogenes in ready-to-eat product, especially sausage samples, using real-time PCR. A standard curve was generated using serially diluted L. monocytogenes cells in distilled water. When cells were artificially inoculated in 10 g of sausage samples in 90㎖ buffered peptone water, the cell concentration of range was approximately 1.0×108 to 100 CFU/㎖. The standard curve of the serially diluted cells was linear for at least seven orders of magnitude from 103 to 109 CFU/㎖ of L. monocytogenes. When cells were diluted in sausages, the linearity range was from 104 to 108 CFU/㎖. The correlation coefficient (R2) of diluted cells was 0.9888 and the slope of the curve was —2.6621. The coefficient and slope of inoculated samples were 0.9916 and —2.747, respectively. The R2 value for serially diluted L. monocytogenes and artificially contaminated sausage samples were acceptable. The approach described in this study represents the potency of the quantification of L. monocytogenes in sausage samples by quantitative real-time PCR. It can be used in monitoring the presence and persistence of this pathogen in sausages.

For the screening of Brucella antibodies in pig, 2,140 pig serum samples were collected from six slaughter house in Korea between 2006 and 2007. The Rose Bengal test (RBT) and serum agglutination test (SAT) were used for initial screening for specific antibodies to Brucella, and competitive enzyme-linked immunosorbent assay (C-ELISA) was used for confirmation of presence of serum antibody for Brucella. Overall, 575 (26.9%) samples resulted in seropositive in RBT. In SAT, 50 (2.3%) and 10 (0.5%) samples showed suspicious positive and positive reaction, respectively, however, all sera tested in this study showed a negative reaction in C-ELISA. SAT and C-ELISA might be applicable as a tool for screening of swine brucellosis.

To determine the prevalence of Campylobacter jejuni and Campylobacter coli in meats, a total of 4,161 samples (1,953 domestic and 2,208 imported) were collected from 304 slaughterhouses nationwide and registered cold storages for imported meats in Korea during 2005～2009. The isolation rates of C. jejuni and C. coli in domestic beef, pork, chicken and duck meats were 0.1% (1/630), 0% (0/630), and 0.1% (1/644), 0% (0/644) and 20.5% (125/609), 10.2% (62/609) and 25.7% (18/70), 20.0% (14/70), respectively. In the case of imported meats, C. jejuni were isolated from 0.1% (1/943) and 15.2% (83/546) of pork and chicken meats, respectively, and C. coli were detected only from 4.8% (26/546) of chicken meats. Neither C. jejuni nor C. coli were detected from imported beef, and C. coli were also not detected from imported pork. In conclusion, chicken meats had much higher rate of contamination with Campylobacter compared to beef and pork. Therefore, HACCP system that is now mandatory for slaughterhouses should be actively practiced for safe and sanitary processing, handling, and marketing of chicken meats. In addition, all critical control points should be determined by processing procedures at processing plants as well as farms and slaughterhouses, and monitoring should be carried out at regular intervals.

Vaginal cytology with behavioral observation was performed in 34 estrous cycles in 16 Miniature Schnauzer dogs to evaluate the its usefulness. The mean duration of proestrus and estrus in Miniature Schnauzer based on behavioral observation and vaginal cytology was (Mean S.D., range: ) and days for proestrus, and and days for estrus, respectively. The duration of each phase of the estrous cycle was not significantly different based on between behavioral observation and vaginal cytology. The gestational length from the first day of male acceptance was days, days from the first male refusal, and days from the onset of cytologic diestrus, respectively. Vaginal cytology during the estrous cycle were significantly characteristic of large intermediate cells in proestrus, anuclear cells in estrus, small intermediate cells in diestrus, and parabasal cells and small intermediate cells in anestrus (p<0.001), respectively. Cornification index (CI) by vaginal cytology was higher in proestrus () and estrus (), then it was decreased in diestrus () and anestrus ().

Serial ultrasonographic examinations were daily performed from 15 days after ovulation until parturition to determine the time of first detection and ultrasonographic appearance of the fetal and extra-fetal structures in pregnant 10 Maltese, 10 Yorkshire Terrier, 15 Shih-tzu, and 10 Miniature Schnauzer bitches, respectively. Gestational age was timed from the day of ovulation (day 0), which was estimated to occur when plasma progesterone concentration was first increased above 4.0ng/ml. The gestational length was (range: ) days and the geatational length was no statistically significant difference among bitches (p>0.05). The initial detection of the extra-fetal structures were; gestational sac at days , zonary placenta at days , yolk sac membrane at days , yolk sac tubular shape at days , and amniotic membrane at days , respectively. The time of the first detection of the extra-fetal structures were no statistically significant difference among bitches (p>0.05). The initial detection of the fetal structures were; embryo initial detection at days , heartbeat at days , embryo bipolar shape , fetal movement at days , limb buds at days , stomach at days , urinary bladder at days , skeleton at days , and kidney at days , respectively. The the time of the first detection of the fetal structures were no statistically significant difference among bitches (p>0.05). These results indicate the evaluation of the time of first detection and ultrasonographic characteristics of the gestational structures might be useful for pregnancy diagnosis, estimating fetal age, embryonic resorption, fetal monster, abnormal fetal growth and fetal viability, respectively.