Interaction of Salt, pH, and Temperature on the Growth and Survival of Salmonellae in Ground Pork

The interaction of temperature, pH, and NaCl concentration on the growth and survival of several strains of salmonellae has been determined in broth and ground pork. Growth of 23 strains occurred in broth at 30 C over a wide range of pH-NaCl combinations; at 10 C, growth was limited to only a few combinations. Cultures which would not grow at 10 C because of the pH-NaCl effect survived for long periods, however. In contrast, cultures which would not grow at 30 C remained viable for only a short time. Results in fresh ground pork were in close agreement with the broth studies. Salmonellae would not grow in ground pork stored at 4 C but would grow in pork containing 3.5% salt stored at 10 C. Salmonellae grew competitively with the natural background flora at 10 C even when the salmonellae constituted less than 5% of the initial flora, and the background flora would grow at a lower temperature than the salmonellae. The data show that, whereas decreasing temperatures increase the inhibitory effects of pH and NaCl, they decrease the lethal effects.     

多疣壁虎的雌性繁殖及孵化温度对孵化期和孵化幼体特征的影响

多疣壁虎 (Ｇｅｋｋｏｊａｐｏｎｉｃｕｓ)是华东地区蜥蜴区系的重要成分 ,向西分布至四川东部 ,向北分布至甘肃和陕西南部 ,国外见于朝鲜南部和日本部分岛屿[7,3 2 ] 。有关该种的生态学研究已涉及 ,①雌雄两性异形和繁殖习性[4 ,16,2 4 ] ;②卵孵化和温度决定性别[3 ,17,2 6,2 7] ;③贮能部位及各部位的年变化和相对重要性评估[1,16] ;④温度对摄食量和食物同化的影响[2 ] ;⑤胚胎和成体的代谢率[6,13 ,3 0 ,3 1] 。浙江杭州产多疣壁虎年产 1～ 3窝卵 ,窝卵数通常恒定为 2枚 ,大个体产多窝卵 ,第 1窝卵和第 2窝卵的卵重无显著差异[4 ] 。…     

Effect of Incubation Temperature on Isolation of Campylobacter jejuni Genotypes from Foodstuffs Enriched in Preston Broth

Preston broth and agar incubated at either 37 or 42°C have been widely used to isolate campylobacters from foodstuffs. The consequences of using either incubation temperature were investigated. Retail packs of raw chicken (n = 24) and raw lamb liver (n = 30) were purchased. Samples were incubated in Preston broth at 37 and 42°C and then streaked onto Preston agar and incubated as before. Two Campylobacter isolates per treatment were characterized. Poultry isolates were genotyped by random amplification of polymorphic DNA (RAPD), pulsed-field gel electrophoresis (PFGE), and flagellin PCR-restriction fragment length polymorphism, and lamb isolates were genotyped by RAPD only. In total, 96% of the poultry and 73% of the lamb samples yielded campylobacters. The lamb isolates were all Campylobacter jejuni, as were 96% of the poultry isolates, with the remainder being Campylobacter lari. The incubation temperature had no significant effect on the number of positive samples or on the species isolated. However, genotyping of the C. jejuni isolates revealed profound differences in the types obtained. Overall (from poultry and lamb), the use of a single incubation temperature, 37°C, gave 56% of the total number of RAPD C. jejuni genotypes, and hence, 44% remained undetected. The effect was especially marked in the poultry samples, where incubation at 37°C gave 47% of the PFGE genotypes but 53% were exclusively recovered after incubation at 42°C. Thus, the incubation temperature of Preston media selects for certain genotypes of C. jejuni, and to detect the widest range, samples should be incubated at both 37 and 42°C. Conversely, genotyping results arising from the use of a single incubation temperature should be interpreted with caution.     

Modeling Colitis-Associated Cancer with Azoxymethane (AOM) and Dextran Sulfate Sodium (DSS)

Individuals with inflammatory bowel disease (IBD), such as Crohn''s disease (CD) or ulcerative colitis (UC) are at increased risk of developing colorectal cancer (CRC) over healthy individuals. This risk is proportional to the duration and extent of disease, with a cumulative incidence as high as 30% in individuals with longstanding UC with widespread colonic involvement.¹ Colonic dysplasia in IBD and colitis associated cancer (CAC) are believed to develop as a result of repeated cycles of epithelial cell injury and repair while these cells are bathed in a chronic inflammatory cytokine milieu.² While spontaneous and colitis-associated cancers share the quality of being adenocarcinomas, the sequence of underlying molecular events is believed to be different.³ This distinction argues the need for specific animal models of CAC.Several mouse models currently exist for the study of CAC. Dextran sulfate sodium (DSS), an agent with direct toxic effects on the colonic epithelium, can be administered in drinking water to mice in multiple cycles to create a chronic inflammatory state. With sufficient duration, some of these mice will develop tumors.⁴ Tumor development is hastened in this model if administered in a pro-carcinogenic setting. These include mice with genetic mutations in tumorigenesis pathways (APC, p53, Msh2), as well as mice pre-treated with genotoxic agents (azoxymethane [AOM], 1,2-dimethylhydrazine [DMH]).⁵The combination of DSS with AOM as a model for colitis associated cancer has gained popularity for its reproducibility, potency, low price, and ease of use. Though they have a shared mechanism, AOM has been found to be more potent and stable in solution than DMH. While tumor development in other models generally requires several months, mice injected with AOM and subsequently treated with DSS develop adequate tumors in as little as 7-10 weeks.^{6, 7} Finally, AOM and DSS can be administered to mice of any genetic background (knock out, transgenic, etc.) without cross-breeding to a specific tumorigenic strain. Here, we demonstrate a protocol for inflammation-driven colonic tumorigenesis in mice utilizing a single injection of AOM followed by three seven-day cycles of DSS over a 10 week period. This model induces tumors with histological and molecular changes closely resembling those occurring in human CAC and provides a highly valuable model for the study of oncogenesis and chemoprevention in this disease.⁸     

Incubation Temperature Effects on Hatchling Performance in the Loggerhead Sea Turtle (Caretta caretta)

Incubation temperature has significant developmental effects on oviparous animals, including affecting sexual differentiation for several species. Incubation temperature also affects traits that can influence survival, a theory that is verified in this study for the Northwest Atlantic loggerhead sea turtle (Caretta caretta). We conducted controlled laboratory incubations and experiments to test for an effect of incubation temperature on performance of loggerhead hatchlings. Sixty-eight hatchlings were tested in 2011, and 31 in 2012, produced from eggs incubated at 11 different constant temperatures ranging from 27°C to 33°C. Following their emergence from the eggs, we tested righting response, crawling speed, and conducted a 24-hour long swim test. The results support previous studies on sea turtle hatchlings, with an effect of incubation temperature seen on survivorship, righting response time, crawling speed, change in crawl speed, and overall swim activity, and with hatchlings incubated at 27°C showing decreased locomotor abilities. No hatchlings survived to be tested in both years when incubated at 32°C and above. Differences in survivorship of hatchlings incubated at high temperatures are important in light of projected higher sand temperatures due to climate change, and could indicate increased mortality from incubation temperature effects.     

Influence of Genotype, Plant Growth Temperature and Anther Incubation Temperature on Microspore Embryo Production in Brassica napus ssp. oleifera

Eleven F₁ hybrid genotypes of winter rape (Brassica napus ssp.oleifera) were used in a study of induction and growth of microspore-derivedembryos. Plants of each genotype were grown in controlled environmentsat either a constant 15°C or a constant 20°C, both witha 16 h photoperiod. Equal numbers of buds, approximately 2.5mm in length, containing uninucleate microspores were harvestedfrom each genotype and either pretreated (14 d at 4°C) ordissected immediately after harvest. Anthers were cultured onliquid medium based upon that of Murashige and Skoog (1962)and containing 8% sucrose, 0.5 mg dm^–3 naphthylaceticacid and 0.05 mg dm^–3 benzylaminopurine. Anthers fromequal samples of buds were incubated at 35°C for 0, 1, 2or 3 d before transfer to 30°C (21 d) and then 25°C.After a total of 42 d incubation, cultures were scored for thepresence of macroscopic embryos (1–2 mm in length) andfor the presence of anthers containing aborted embryoids whichhad not developed further. The results showed first that bud pretreatment completely inhibitedinduction and secondly that anthers of all genotypes had anabsolute requirement for a 35°C treatment (optimal duration2 d) in order to induce embryoid formation. In the great majorityof genotypes plants grown at 15°C provided more productiveanthers than plants grown at 20°C. However, within eachtreatment there were great differences both in the frequencyof anthers showing induced embryoids and of the final yieldof embryos. There was evidence that hybrids with a common parentresponded similarly under certain treatments. This confirmedthe importance of genotypic control for some components of embryoyield. Key words: Brassica napus, Rape, Anther culture, Pollen, Haploid     

Cyclic-AMP Production by Corynebacterium murisepticum No. 7 (ATCC 21374) and Microbacterium sp. No. 205 (ATCC 21376)

Corynebacterium murisepticum No. 7 (ATCC 21374) and Microbacterium sp. No. 205 (ATCC 21376) isolated from soil by the authors did not require purines for their growth and accumulated a large amount of cyclic-AMP (7~12mmoles) in the cultural medium, supplemented with adenine, hypoxanthine or their derivatives, but scarcely accumulated without these compounds, and d,l-alanine as a sole nitrogen source was not effective on cyclic-AMP production without these compounds

Four to five percent of glucose were the most effective on the production. Yeast extract and poly peptone were required for cyclic-AMP production.     

The Effects of Temperature on the Incubation and Latent Periods of Powdery Mildew (Erysiphe polygoni) on Clematis

The effects of temperature on the length of the incubation and latent periods of clematis powdery mildew, caused by Erysiphe polygoni , were studied. At constant temperatures over the range of 10–29°C, the length of the incubation period ranged from 4 to 11 days, and the length of the latent period ranged from 4 to 13 days; no visible colonies developed at 30°C after 19 days. The relationship between temperature and the rates of fungal development within the incubation and latent periods (expressed as the reciprocal of the lengths of the incubation and latent periods) under constant temperature were described well by a non-linear model. The resulting curves were not the usual form of an asymmetrically bell-shaped type; instead, they were of exponential type with the development rate increasing with increasing temperatures. The relationship between temperature and the development rate during the post-incubation (before sporulation) period was non-linear with an optimum temperature of approximately 20°C.     

Influence of Temperature of Incubation and Type of Growth Medium on Pigmentation in Serratia marcescens

Maximal amounts of prodigiosin were synthesized in either minimal or complete medium after incubation of cultures at 27 C for 7 days. Biosynthesis of prodigiosin began earlier and the range of temperature for formation was greater in complete medium. No prodigiosin was formed in either medium when cultures were incubated at 38 C; however, after a shift to 27 C, pigmentation ensued, provided the period of incubation at 38 C was not longer than 36 hr for minimal medium or 48 hr for complete medium. Washed, nonpigmented cells grown in either medium at 38 C for 72 hr could synthesize prodigiosin when suspended in saline at 27 C when casein hydrolysate was added. These suspensions produced less prodigiosin at a slower rate than did cultures growing in casein hydrolysate at 27 C without prior incubation at 38 C. Optimal concentration of casein hydrolysate for pigment formation by suspensions was 0.4%; optimal temperature was 27 C. Anaerobic incubation, shift back to 38 C, killing cells by heating, or chloramphenicol (25 mug/ml) inhibited pigmentation. Suspensions of washed cells forming pigment reached pH 8.0 to 8.3 rapidly and maintained this pH throughout incubation for 7 days. Measurements of viable count and of protein, plus other data, indicated that cellular multiplication did not occur in suspensions of washed cells during pigment formation. By this procedure utilizing a shift down in temperature, biosynthesis of prodigiosin by washed cells could be separated from multiplication of bacteria.     

Isolation and characterization of the 7S protein from glanded cottonseed (Gossypium herbacium)

The major protein from glanded cottonseed has been isolated in a homogeneous form. Its S²⁰ w value at 1 protein concentration is 6S in 1 M NaCl solution. It contains 1 carbohydrate and is free from phosphorus, gossypol (bound or free) and nucleic acid impurities. It consists of atleast seven non-identical subunits. The protein has an ultraviolet absorption maximum at 278 nm and fluorescence excitation and emission maxima at 280 nm and 325 nm respectively. Optical rotatory dispersion and circular dichroism measurements indicate that the protein consists predominantly of Β-structure and random coil. The observed near-ultraviolet circular dichroic bands can be attributed to tyrosine, phenylalanine and tryptophan residues of the protein.     

Influence of Incubation Temperature on the Microbial Reductive Dechlorination of 2,3,4,6-Tetrachlorobiphenyl in Two Freshwater Sediments

We studied the impact of incubation temperatures on the dechlorination of 2,3,4,6-tetrachlorobiphenyl (2346-CB) in two sediments from different climates: polychlorinated biphenyl (PCB)-free sediment from Sandy Creek Nature Center Pond (SCNC) in Athens, Ga., and PCB-contaminated sediment from Woods Pond (WP) in Lenox, Mass. Sediment slurries were incubated anaerobically with 350 (mu)M 2346-CB for 1 year at temperatures ranging from 4 to 66(deg)C. Most of the 2346-CB was dechlorinated between 12 and 34(deg)C in both sediments and, unexpectedly, between 50 and 60(deg)C in WP sediment. This is the first report of PCB dechlorination at thermobiotic temperatures. The data reveal profound differences in dechlorination rate, extent, and products as a function of sediment and temperature. The highest observed rate of dechlorination of 2346-CB to trichlorobiphenyls occurred at 30(deg)C in both sediments, but the rate was higher for WP than for SCNC sediment (46 versus 16 (mu)mol liter(sup-1) day(sup-1)). For SCNC sediment the rate of dechlorination dropped sharply below 30(deg)C, but for WP sediments it was near optimal from 20 to 34(deg)C and then dropped sharply below 20(deg)C. In WP sediment most of the meta chlorines were removed between 8 and 34(deg)C and between 50 and 60(deg)C. para dechlorination was restricted from 18 to 34(deg)C and was optimal at 20(deg)C. ortho dechlorination occurred between 8 and 30(deg)C, with optima around 15 and 27(deg)C, but the extent was highly variable. In SCNC sediment complete meta dechlorination occurred from 12 to 34(deg)C and para dechlorination occurred from 18 to 30(deg)C; both were optimal at 30(deg)C. No ortho dechlorination was observed. Dechlorination products were 246-CB, 236-CB, and 26-CB (both sediments) and 24-CB, 2-CB, and 4-CB (WP sediment). The data suggest that in SCNC sediment similar factors controlled meta and para PCB dechlorination over a broad temperature range (18 to 30(deg)C) but that in WP sediment there were multiple temperature-dependent changes in the factors controlling ortho, meta, and para dechlorination. We attribute the differences observed in the two sediments to differences in their PCB-dechlorinating communities.     

孵化温度对灰鼠蛇卵孵化期、孵化成功率和孵出幼体特征的影响

用4个恒定温（24－32℃）孵化灰鼠蛇卵,检测温度对孵化期,孵化成功率和孵出幼体特征的影响。在24－32℃范围内,孵化温度显影响孵化期及孵出幼体的体长和剩余卵黄大小,但不影响孵化成功率和孵出幼体的性别,体重,躯干重和脂肪体重。24,26,30和32℃孵化期分别为99．0,72．2,54．7和48．7d。24℃和26℃孵出幼体的体筮大于30℃和32℃孵出幼体;24℃和32℃孵同幼体内的卵黄较多。不同温度下发育的胚胎对卵内物质和能量的利用一定的差异,但差异不显。雌性幼体的体长,尾长和总长均大于雄性幼体,这些两性差异与孵化温度无关。孵出幼体和新生卵内容的灰分含量无显差异,孵化前后卵壳灰分含量也无显差异,表明灰鼠蛇的卵黄可提供胚胎发育所需的所有无机物。     

The Effect of Male Incubation Feeding,Food and Temperature on the Incubation Behaviour of New Zealand Robins

Because of finite resources, organisms face conflict between their own self‐care and reproduction. This conflict is especially apparent in avian species with female‐only incubation, where females face a trade‐off between time allocated to their own self‐maintenance and the thermal requirements of developing embryos. We recorded incubation behaviour of the New Zealand robin (Petroica longipes), a species with female‐only incubation, male incubation feeding and high nest predation rates. We examined how male incubation feeding, ambient temperature and food availability (invertebrate biomass) affected the different components of females’ incubation behaviour and whether incubation behaviour explained variation in nest survival. Our results suggest that male incubation feeding rates of 2.8 per hour affect the female’s incubation rhythm by reducing both on‐ and off‐bout duration, resulting in no effect on female nest attentiveness, thus no support for the female‐nutritional hypothesis. The incubation behaviours that we measured did not explain nest survival, despite high nest predation rates. Increased ambient temperature caused an increase in off‐bout duration, whereas increased food availability increased on‐bout duration. While males play a vital role in influencing incubation behaviour, female robins attempt to resolve the trade‐off between their own foraging needs and the thermal requirements of their developing embryos via alternating their incubation rhythm in relation to both food and temperature.     

Isolation of Salmonellae from Foods Samples: V. Determination of the Method of Choice for Enumeration of Salmonella

Comparison of three methods by which salmonellae may be isolated and enumerated from dried albumen, direct inoculation of enrichment media, centrifugation of samples, and pre-enrichment in noninhibitory media, reveals pre-enrichment to be the method of choice.

The superiority of pre-enrichment manifests itself in replicate aliquots of the same sample by producing a statistically significant increase in numbers of isolations of salmonellae and in empirical use with various albumen samples by consistently higher values of most probable numbers (MPN).

The primary factor involved in this superiority appears to be the greater ability of small numbers of salmonellae to initiate growth in the nonselective mannitol purple sugar broth than in the inhibitory enrichment media.

The method of analysis recommended entails inoculation of mannitol broth pre-enrichment medium, transfer of 24-hr culture aliquots to tetrathionate broth, and streaking on brilliant green agar for isolation of salmonellae.