OPTIMIZATION OF THE ENUMERATION OF TOTAL AEROBIC BACTERIAL FLORA IN THE FLESH OF SEAFISH

Abstract The total aerobic flora of seafish flesh is weakly halophilic, and requires on average 1.38% NaCl according to statistical studies. Enumeration is optimal on tryptone soya agar or on NaCl supplemented plate count agar (-H₂S), incubated at 20 and 25C, respectively. Plate count agar (-H₂S) was selected because it can also be used for enumeration of hydrogen sulfide-producing bacteria by degradation of sulfur-containing proteins, which are abundant in fish The models employed are sigmoidal. The initial bioburden is too great for there to be a lag phase during storage in ice at 0C. The models show that when the total aerobic microflora count exceeds 100,000 cfu/g, whole or filleted fish stored on ice at 0C are unfit for consumption.     

OPTIMIZATION OF CULTURE CONDITIONS FOR ENUMERATION OF H2S BACTERIA IN THE FLESH OF SEAFISH

H₂S bacteria of seafish flesh are weakly halophilic and require on average 1.68% NaCl according to statistical studies. Enumeration is optimal on PCA-H₂S(a PCA medium supplemented with sulfur sources and increased NaCl concentrations) incubated at 25C. Total aerobic bacteria can be counted simultaneously on this medium. The proportion of H₂S bacteria relative to total aerobic bacteria increased slightly during prolonged storage of the fish, but was highly variable. Models relating H₂S bacterial counts to spoilage of fish are sigmoidal and showed that when the count exceeds 10,000 CFU/g, whole or filleted fish stored in ice at 0C are unfit for consumption. Shewanella putrefaciens accounted for 69% of the H₂S bacteria at the fifth day of storage and 100% at the fifteenth.     

A single chicken anemia virus protein induces apoptosis.

Chicken anemia virus (CAV) causes cytopathogenic effects in chicken thymocytes and cultured transformed mononuclear cells via apoptosis. Early after infection of chicken mononuclear cells, the CAV-encoded protein VP3 exhibits a finely granular distribution within the nucleus. At a later stage after infection, VP3 forms aggregates. At this point, the cell becomes apoptotic and the cellular DNA is fragmented and condensed. By immunogold electron microscopy VP3 was shown to be associated with apoptotic structures. In vitro, expression of VP3 induced apoptosis in chicken lymphoblastoid T cells and myeloid cells, which are susceptible to CAV infection, but not in chicken embryo fibroblasts, which are not susceptible to CAV. Expression of a C-terminally truncated VP3 induced much less pronounced apoptosis in the chicken lymphoblastoid T cells.     

Enhanced reactivation and enhanced mutagenesis of herpes simplex virus in normal human and xeroderma pigmentosum cells.

Enhanced reactivation (ER) and enhanced mutagenesis (EM) of herpes simplex virus type 1 were studied simultaneously in UV-irradiated stationary cultures of diploid normal human and xeroderma pigmentosum (XP) fibroblasts. Mutagenesis was assayed with unirradiated herpes simplex virus type 1 as a probe in a forward mutation assay (resistance to iododeoxycytidine). Dose-response studies showed that ER increased with the UV dose given to the virus. Optimal reactivation levels were obtained when normal cells and XP variant cells were exposed to a UV dose of 8 J . m-2 and the virus was irradiated with 150 J . m-2. Repair-deficient XP cells of complementation groups A, C, and D showed optimal reactivation levels with a UV dose to the cells of 1.0 J . m-2 and a UV dose to the virus of 40 J . m-2. The time course of appearance of ER and EM was also studied, both in the normal and XP cells. In all cell types except the XP variant cells, EM followed similar kinetics of appearance as did ER. Maximal activities occurred when infection was delayed 1 or 2 days after cell treatment. In XP variant cells, however, maximal expression of the EM function was significantly delayed with respect to ER. The results indicate that ER and EM are transiently expressed in normal and repair-deficient XP cells. Although both phenomena may be triggered by the same cellular event, ER and EM appear to be separate processes that occur independently of each other.     

On the problem of radioadaptation in small mammals (ecological specialization of species, radioresistance, hemopoiesis, immunity)

The data illustrating ambivalent nature of radioresistance of rodents having different ecological specialization: wood mice (Apodemus uralensis Pallas 1811) and mole-voles (Ellobius talpinus Pallas 1770) to acute (laboratory experiment) and chronic (long-term inhabiting the East Ural Radioactive Trace - EURT) radiation action are presented. These species are greatly distinguished by a way of life, a migratory activity, an average life span etc. Numerous immunological, hematological disturbances were found in more radioresistant wood mice inhabiting in radioactively-polluted biogeocenosis (density of pollution 90Sr - 18.5 MBq/m2 = 500 Ci/km2). On the contrary there are no pathological shifts in more radiosensitive mole-voles from the EURT zone (density of pollution 90Sr - 37 MBq/m2 = 1000 Ci/km2). Comparative analysis of the hematological and immunological parameters and absorbed dose loads in rodents from radiocontaminated environment testify about the possibility of radioadaptation in a series of generations in Ellobius talpinus Pall. The leading role in development of radioadaptation of mole-voles belongs to ecophysiological peculiarities of the species.     

An Ex Vivo Study of Congenital Pigmented Nevi in Epidermal Reconstructs

In order to study morphologic and functional characteristics of pigment cells in congenital pigmented nevi, autologous or heterologous reconstructs have been made using normal keratinocytes and nevus cells from the dermal-epidermal junction or from the dermis. All these cells, keratinocytes and nevus cells, were used as cell suspensions immediately after dissociation from the tissues or after subsequent brief cultivation in a serum-free medium. Reconstructed epidermis were cultured for 15 days at the air-liquid interface with or without ultraviolet (UV) B exposure. The reconstructs were examined macroscopically (formation of hyperpigmented macules), histologically (pigment cell nesting) and ultrastructurally (pigment structure and transfer). Typical nesting of nevus cells was observed in the dermal-epidermal junction or in the superficial dermis associated with macroscopically detectable small pigmented macules. UVB exposure induced an upward migration of nevus cells in the suprabasal layers of the epidermis. This tissue model can be considered as an excellent system for the ex vivo reproduction of pigmented nevi and as an assay of the sensitivity of nevus cells towards UVB irradiation.     

Higher number of <Emphasis Type="Italic">Helicobacter pylori</Emphasis> CagA EPIYA C phosphorylation sites increases the risk of gastric cancer,but not duodenal ulcer

Background  

Helicobacter pylori infection is one of the most common infections worldwide and is associated with gastric cancer and peptic ulcer. Bacterial virulence factors such as CagA have been shown to increase the risk of both diseases. Studies have suggested a causal role for CagA EPIYA polymorphisms in gastric carcinogenesis, and it has been shown to be geographically diverse. We studied associations between H. pylori CagA EPIYA patterns and gastric cancer and duodenal ulcer, in an ethnically admixed Western population from Brazil. CagA EPIYA was determined by PCR and confirmed by sequencing. A total of 436 patients were included, being 188 with gastric cancer, 112 with duodenal ulcer and 136 with gastritis.     

Adaptation of small mammals to radiocontaminated environment: Oxidative metabolism and radioresistance

Differences in radioresistance and oxidative metabolism parameters in rodents of different ecological specialization in the East Ural radioactive trace (EURT) area are shown. Northern mole voles (radiosensitive species) exhibit a pronounced resistance to chronic exposure to radiation, living in a series of generations in a radioactive environment, whereas pigmy wood mice (radioresistant species) are less resistant in the EURT area due to their migration activity. Thus, under the EURT area conditions, northern mole voles realize the genotypic radioadaptation, whereas pigmy wood mice realize the physiological radioadaptation, the key role in which is played by the ecological and physiological characteristics and the lifestyle of animals.