The influence of plating technique and incubation temperature on bacterial count from fish and fishery products

For bacterial sampling of raw unprocessed fish and frozen fishery products, spread plate method is preferable to pour plate method; incubation of plates at 30°C gives a higher count than incubation at 37°C. Analysis of variance of the data shows that sample variation between different types of fishes is highly significant whereas the variations between triplicate plates are not significant at 5 % level.

Influence of exogenous fibrolytic enzyme level and incubation pH on the in vitro ruminal fermentation of alfalfa stems

A series of in vitro experiments was carried out to examine the impact of enzyme application rate and incubation medium pH on the rate and extent of fermentation of alfalfa stems. In Experiment 1, a commercial enzyme product (Liquicell 2500, Specialty Enzyme and Biochemicals, Fresno, CA, USA) was added to alfalfa stems at six levels: 0, 0.51, 1.02, 2.55, 5.1, and 25.5 mu l/g (control and L1-L5, respectively) to forage DM in a completely randomized design, with a factorial arrangement of treatments. Rate and extent of fermentation and apparent organic matter degradation (OMD) were determined in vitro, using a gas production technique. Addition of enzyme linearly increased (P < 0.01) gas production for up to 12 h (68.9, 70.9, 67.6, 67.9, 71.9, and 74.9 ml/g OM for control, L1-L5, respectively) and OMD for up to 19 h incubation (0.425, 0.444, 0.433, 0.446, 0.443, and 0.451 for control, L1-L5, respectively), but no increases (P > 0.05) were detected thereafter. In Experiment 2, the effect of the same enzyme as used previously (added at 0.51 mu l/g forage DM, directly into the incubation medium), and buffer pH were examined using the ANKOM system, in a completely randomized design. Incubation medium pH was altered using 1 M citric acid, in order to obtain target initial pH values of 6.8 (control, no citric acid added), 6.2, 5.8, and 5.4. Actual initial pH values achieved were 6.72, 6.50, 6.20, and 5.72. Lowering the pH decreased (P < 0.01) dry matter disappearance (DMD) at 18 h incubation (0.339, 0.341, 0.314, and 0.291 for 6.72, 6.50, 6.20, and 5.72, respectively), whereas enzyme addition increased (P < 0.05) DMD at 24 h (0.363 versus 0.387 for control and enzyme-treated, respectively). Addition of enzyme increased (P < 0.05) neutral detergent fibre (NDF), acid detergent fibre (ADF), and hemicellulose (HC) degradation at pH 6.50 (0.077 versus 0.117; 0.020 versus 0.051; 0.217 versus 0.270 for control and enzyme-treated NDF, ADF and hemicellulose degradation, respectively) and 6.72 (0.091 versus 0.134; 0.041 versus 0.079; 0.205 versus 0.261 for control and enzyme-treated NDF, ADF and HC degradation, respectively). It is concluded that the positive effects of this enzyme product were independent of the pre-treatment period, but pH influenced the responses to enzyme supplementation. Under the conditions of this experiment, exogenous fibrolytic enzymes seemed to work better at close to neutrality ruminal pH conditions. (C) 2006 Elsevier B.V. All rights reserved.

Isolation and mycelial growth of Diehliomyces microsporus: effect of culture medium and incubation temperature

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Viability of Streptococcus mutans on transparent and opaque toothbrushes.

PURPOSE: The objective of this study was to evaluate the viability of the microorganism Streptococcus mutans on toothbrushes made of opaque and transparent materials. METHODS: Twenty-eight toothbrushes (14 opaque and 14 transparent) were inoculated in tubes with brain heart infusion (BHI) broth of a standard strain of S. mutans and incubated in candle jars at 37 degrees C for 24 hours. Both the opaque and transparent toothbrushes were removed at T = 0 h (control); T = 0.5 h; T = 1 h; T = 2 h; T = 4 h; T = 8 h; and T = 24 h. Individual toothbrushes were subjected to agitation in a saline solution and samples of the solution were diluted and inoculated in Bacitracin Sucrose Agar--SB-20. RESULTS: After half an hour (T2) there was a significant decrease in the number of microorganisms on the transparent and opaque toothbrushes, respectively 6.0 x 10(5) and 9.4 x 10(5), when compared to the control. After the T3 = 1 hour, T4 = 2 hours, T5 = 4 h, the number of microorganisms decreased from 4.1 x 10(5); 2.1 x 10(5); 1.4 x 10(5); and 9.2 x 10(5); 5.7 x 10(5); 1.2 x 10(5) to zero (0.0) in T6 = 8 h, respectively on the transparent and opaque toothbrushes. The reduction in viable microorganisms was more obvious with the transparent toothbrushes, although the number of viable microorganisms was not significantly different for the two types of toothbrushes at the end of the experiment, T5 = 1.4 x 10(5) (transparent) and T5 = 1.2 x 10(5) (opaque). CONCLUSIONS: With both opaque and transparent toothbrushes, the number of microorganisms decreased with time. A reduction in the number of microorganisms on the transparent toothbrushes was observed following inoculation and incubation. This suggests the transparent toothbrushes inhibit the viability of the S. mutans.

Hematological and incubation parameters of chicks from young breeders eggs: Variation with sex and incubation temperature

This experiment analyzed the effect of sex and incubation temperature on daily mass loss and eggshell conductance, embryo mortality rates, incubation duration, hematological parameters and body, liver, heart and bursa weights of neonatal chicks from young breeders. The daily mass loss was higher at incubation temperature of 39°C. The eggshell conductance rate increased with the temperature. The total and partial duration of incubation were lower for eggs incubated at 39°C. The time taken by the chick to leave the eggshell did not differ below and above the thermoneutral temperature. The total and intermediate embryo mortality rates increased with the incubation temperature, whereas the early and late embryo mortality rates were higher at incubation temperature of 39°C. Sex did not influence the analyzed parameters, while the incubation temperature did not affect the body and bursa weight and the erythrocytes characteristics. The liver weight of chicks incubated at 36°C was higher than the incubated at 39°C, however there were no differences among the liver weight from chicks incubated at 36 and 39°C and those incubated at 37.5°C. The number of heterophils and the heterophil/lymphocyte ratio (H/L ratio) increased following the temperature, whereas the number of lymphocytes decreased at high temperatures. The other leukocyte parameters did not suffer influence of temperature. Males and females presented similar response to variation of incubation temperatures (36, 37.5 and 39°C) and demonstrated higher sensibility to temperatures above the thermoneutral. Moreover, temperatures below the thermoneutral demonstrated to be better for improvement of hatchability and development of chicks from light eggs. © Asian Network for Scientific Information, 2010.

Efficacy of sperm motility after processing and incubation to predict pregnancy after intrauterine insemination in normospermic individuals

Background: Intrauterine insemination (IUI) is widely used to treat infertility, and its adequate indication is important to obtain good pregnancy rates. To assess which couples could benefit from IUI, this study aimed to evaluate whether sperm motility using a discontinuous gradient of different densities and incubation in CO2 in normospermic individuals is able to predict pregnancy.Methods: A total of 175 couples underwent 175 IUI cycles. The inclusion criteria for women were as follows: 35 years old or younger (age range: from 27 to 35 years) with normal fallopian tubes; endometriosis grades I-II; unexplained infertility; nonhyperandrogenic ovulatory dysfunction. Men with normal seminal parameters were also included. All patients underwent ovarian stimulation with clomiphene citrate and human hMG or r-FSH. When one or (at most) three follicles measuring 18 to 20 mm were observed, hCG (5000 UI) or r-hCG (250 mcg) was administered and IUI performed 36-40 h after hCG. Sperm processing was performed using a discontinuous concentration gradient. A 20 microliters aliquot was incubated for 24 h at 37 degrees C in 5% CO2 following a total progressive motility analysis. The Mann-Whitney and Chi-square tests, as well as a ROC curve were used to determine the cutoff value for motility.Results: Of the 175 couples, 52 (in 52 IUI cycles) achieved clinical pregnancies (CP rate per cycle: 29.7%). The analysis of age, duration and causes of infertility did not indicate any statistical significance between pregnancy and no pregnancy groups, similar to the results for total sperm count and morphology analyses, excluding progressive motility (p < 0.0001). The comparison of progressive motility after processing and 24 h after incubation between these two groups indicated that progressive motility 24 h after incubation was higher in the pregnancy group. The analysis of the progressive motility of the pregnancy group after processing and 24 h after incubation has not shown any motility difference at 24 h after incubation; additionally, in couples who did not obtain pregnancy, there was a statistically significant decrease in progressive motility 24 h after incubation (p < 0.0001). The ROC curve analysis generated a cutoff value of 56.5% for progressive motility at 24 h after incubation and this cutoff value produced 96.1% sensitivity, 92.7% specificity, 84.7% positive predictive value and 98.3% negative predictive value.Conclusions: We concluded that the sperm motility of normospermic individuals 24 h after incubation at 37 degrees C in 5% CO2, with a cutoff value of 56.5%, is predictive of IUI success.

Effect of diazotrophic bacterium inoculation and organic fertilization on yield of Champaka pineapple intercropped with irrigated sapota.

The purpose of this study was to evaluate the response of the Champaka pineapple to inoculation with the diazotrophic bacterium Asaia bogorensis (strain 219) when grown with organic fertilizer in an irrigated sapota orchard. Plantlets were transplanted to tubes containing a mixture of worm compost and vermiculite and inoculated with 108 bacterial cells. After five and a half months of acclimatization the plantlets were transplanted in furrows in the sapota orchard. Fertilizer was placed at the bottom of the furrows and covered with three doses (2.5; 5.0 and 7.5 L linear m−1 row) of three organic composts. The successful association of the plantlets with the diazo-trophic bacterium was confirmed by most probable number analysis before transferring to the field. Plants inoculated with strain AB219 showed the greatest initial leaf growth and produced the heaviest fruits compared to uninoculated plants. Plant growth and fruit yield increased with increasing compost dosages. The results suggested that Champaka pineapple benefited from the association of A. bogorensis (strain 219) when grown under irrigation and with organic fertilizer.

L-Glutaminase production by marine vibrio costicola under solid state fermentation

Use of inert supports have been recommended for SSF in on ar to overcome its inherent problems and efforts are being made to search for newer and better materials to act as inert solid supports lidoo et al, 1982; Zhu et al, 1994).In the present study an attempt is made to produce L-glutaminase, which is industrially and therapeutically impo rtant, from marine bacteria under solid state fermentation using natura.l. inert and mixed substrates with a view to develop an ideal bioprocess for its large scale production.

The nature of innate and adaptive interleukin-17A responses in sham or bacterial inoculation

Streptococcus pyogenes is the causative agent of numerous diseases ranging from benign infections (pharyngitis and impetigo) to severe infections associated with high mortality (necrotizing fasciitis and bacterial sepsis). As with other bacterial infections, there is considerable interest in characterizing the contribution of interleukin-17A (IL-17A) responses to protective immunity. We here show significant il17a up-regulation by quantitative real-time PCR in secondary lymphoid organs, correlating with increased protein levels in the serum within a short time of S. pyogenes infection. However, our data offer an important caveat to studies of IL-17A responsiveness following antigen inoculation, because enhanced levels of IL-17A were also detected in the serum of sham-infected mice, indicating that inoculation trauma alone can stimulate the production of this cytokine. This highlights the potency and speed of innate IL-17A immune responses after inoculation and the importance of proper and appropriate controls in comparative analysis of immune responses observed during microbial infection.

Mobile Incubation in Waved Albatross (Phoebastria irrorata): Associated Hatching Failure and Artificial Mitigation

Waved albatrosses often relocate their eggs during incubation by placing the egg between the tarsi and shuffling forward. This behavior frequently results in eggs becoming lodged between rocks, accounting for at least 10%, and perhaps as much as 80%, of breeding failures. Because albatross populations worldwide are currently threatened, artificial means of augmenting reproductive success may be necessary to mitigate losses caused by anthropogenic effects. We characterize the frequency and extent of egg movement; test several hypotheses related to microhabitat, timing, and incubation location to explain the behavior; and investigate the utility of repositioning lodged eggs in a location in which breeding birds might resume incubation. Egg rescue increased both the likelihood of continued incubation as well as the hatching rate in our experiment, and provides an efficient, low-cost management option for this species.

Immunisation of dogs, hamsters and guinea pigs against Rhipicephalus sanguineus using crude unfed adult tick extracts

Naive experimental groups of dogs, hamsters and guinea pigs were inoculated three times subcutaneously with unfed adult extract of the tick Rhipicephalus sanguineus and challenged with adult R. sanguineus to evaluate resistance. The acquisition of resistance was based on alterations of some reproductive and feeding performance parameters of female ticks such as female and egg mass weights, engorgement, pre-oviposition and incubation periods, larval hatchability rate and efficiency rates of female ticks in converting their food reservoir to eggs and larvae. Dogs did not develop resistance under these experimental conditions; guinea pigs and hamsters, to a lesser extent, acquired an effective immunity to ticks as demonstrated by the impairment of the reproductive and feeding performance. However, the resistance induced by inoculation of the extract in the rodents seemed not to be as efficient as that induced by successive infestations.

Comparison of brain PrPd distribution in ovine BSE and scrapie

Scrapie and bovine spongiform encephalopathy (BSE) are both prion diseases affecting ruminants, and these diseases do not share the same public health concerns. Surveillance of the BSE agent in small ruminants has been a great challenge, and the recent identification of diverse prion diseases in ruminants has led to the development of new methods for strain typing. In our study, using immunohistochemistry (IHC), we assessed the distribution of PrP(d) in the brains of 2 experimentally BSE-infected sheep with the ARQ/ARQ genotype. Distribution of PrP(d) in the brain, from the spinal cord to the frontal cortex, was remarkably similar in the 2 sheep despite different inoculation routes and incubation periods. Comparatively, overall PrP(d) brain distribution, evaluated by IHC, in 19 scrapie cases with the ARQ/ARQ, ARQ/VRQ, and VRQ/VRQ genotypes, in some cases showed similarities to the experimentally BSE-infected sheep. There was no exclusive neuroanatomical site with a characteristic and specific PrP(d) type of accumulation induced by the BSE agent. However, a detailed analysis of the topography, types, and intensity of PrP(d) deposits in the frontal cortex, striatum, piriform cortex, hippocampus, mesencephalon, and cerebellum allowed the BSE-affected sheep group to be distinguished from the 19 scrapie cases analyzed in our study. These results strengthen and emphasize the potential interest of PrP(d) brain mapping to help in identifying prion strains in small ruminants.