Effect of Dimethyl Sulfoxide on Cell Cycle Synchronization of Goldfish Caudal Fin Derived Fibroblasts Cells

Several studies have previously been conducted regarding cell cycle synchronization in mammalian somatic cells. However, limited work has been performed on the control of cell cycle stages in the somatic cells of fish. The aim of this study was to determine the cell cycle arresting effects of several dimethyl sulfoxide (DMSO) concentrations for different times on different cell cycle stages of goldfish caudal fin‐derived fibroblasts. Results demonstrated that the cycling cells or control group (68.29%) yields significantly higher (p < 0.05) arrest in G0/G1 phase compared with the group treated for 24 h with different concentrations (0.5%, 1.0% or 1.5%) of DMSO (64.88%, 65.70%, 64.22% respectively). The cell cycle synchronization in the treatment of cells with 1.0% DMSO at 48 h (81.14%) was significantly higher than that in the groups treated for 24 h (76.82%) and the control group (77.90%). Observations showed that treatment of DMSO resulted in an increase in the proportion of cells at G0/G1 phase for 48 h of culture. However, high levels of apoptotic cells can be detected after 48 h of culture treated with 1% concentration of DMSO.     

Infiltrating Foxp3+ Regulatory T cells and Histopathological Features in Canine Classical and Spermatocytic Seminomas

In humans, regulatory T (T reg) cells are known to play a critical role in both the regulation of immune homoeostasis and the progression of cancer. However, there is little information about the identification, characterization and the function of T reg cells in canine tumours. We identified T reg cells in 28 canine seminoma samples using a Forkhead box P3 (Foxp3) antibody and investigated the relationship between T reg cell infiltration and histopathological features of classical and spermatocytic seminomas (SE and SS, respectively). The Foxp3 protein showed nuclear immunostaining in infiltrating lymphocytes, and Foxp3+ cells were diffused or focally distributed in seminoma tissues. Foxp3+ cells were frequently present in the SS histotype, in seminomas that showed no evidence of tumour cell invasion into the vessels and in seminomas showing a diffuse growth pattern with three cell types. Neither the SE/SS histotype nor the histopathological features of the tumour correlated with Foxp3+ cell counts. These results indicate that Foxp3+ T reg cells may be associated with a less malignant histological phenotype or may not play a critical role in the immune response of canine seminomas. Moreover, Foxp3+ T reg cells may be associated with SS seminoma, but further studies, involving a larger number of samples, are required to better understand whether these cells play a critical role in the immune response in canine seminomas. This is the first report to demonstrate the characteristics of T reg cell infiltration in canine seminoma.     

The introduction of expert systems in animal husbandry

The intensive nature of modern animal husbandry has changed the environmental conditions, so that protection against diseases, especially those caused by secondary pathogenic organisms, becomes an increasing concern. Production inefficiencies become more and more important. Because of their complex nature, many associated problems cannot be solved by farmers alone. With the introduction of information technology in the form of expert systems, new possibilities emerge for maximal productivity and animal health. Expert systems are a new type of computer programmes which are able to handle problems in limited domains at an expert level, using expert knowledge and reasoning processes, within strictly defined problem limits. Expert systems as they presently exist have the following components: a knowledge base (rule- and data-base), inference engine (deductive mechanism), user-interface (explanatory facilities, etc.). The application of knowledge engineering in animal husbandry may result in improvements in the general health and productivity of the herd and the financial returns of the farm, derived from the improved managerial control. At the same time the use of knowledge engineering may lead to greater understanding of the various epidemiological factors that influence farm results, either positively or negatively.     

Estimating the incidence of influenza-virus infections in Dutch weaned piglets using blood samples from a cross-sectional study

A cross-sectional study was carried out on 32 Dutch breeding herds to estimate the incidence of influenza-virus infections in piglets before the start of the finishing period, at the age of approximately 10 weeks. Longitudinal studies on two herds (8 and 10 litters, respectively) were done to obtain an average decay function for maternal antibodies.Each participating farm in the cross-sectional study was visited twice within 5 months; each time, blood samples were taken randomly from one compartment (a separate room with separate air flow) of 4-5-week-old piglets and one compartment of 8-9-week-old piglets. These blood samples (a total of 2598; 16-23 per compartment, depending on its size) were tested in a haemagglutination inhibition test for antibodies against influenza-virus subtypes H1 and H3. Samples from 8-9-week-old piglets from the first sampling period (n=660) were also tested in an IgM ELISA.For each individual herd and each influenza-virus subtype separately, the decay function derived from the longitudinal studies was used to calculate an expected seroprevalence in 8-9-week-old piglets, which was then compared to the observed seroprevalence. Depending on subtype and sampling period, between 10 and 15 of the 32 herds were suspected of virus circulation during the weaning period because the observed seroprevalence was significantly higher than the expected seroprevalence (P<0.05). In the first sampling period the IgM ELISA confirmed six of these outbreaks. However, due to the small window of detection of the IgM ELISA (compared to the length of the weaning period), it will always underestimate the number of infections. Infections in the first half of the weaning period will no longer be detectable because IgM antibodies have already disappeared.In individual pigs, an incidence of 16-17% was estimated for each subtype over a 4-week period between the age of 4-5 and 8-9 weeks. For each influenza subtype, 80% of the piglets will enter the finishing facilities without antibodies or with decaying maternal antibodies. These piglets may be susceptible to an infection with influenza virus.     

Molecular Basis of Gene-for-Gene Specificity in Bacterial Speck Disease of Tomato

Transient expression of the Pseudomonas syringae avirulence gene avrPto in plant cells resulted in a Pto-dependent necrosis. The AvrPto avirulence protein was observed to interact directly with the Pto resistance protein in the yeast two-hybrid system. Mutations in the Pto and avrPto genes which reduce in vivo activity had parallel effects on association in the two-hybrid assay. These data suggest that during infection the pathogen delivers AvrPto into the plant host cell and that resistance is specified by direct interaction of Pto with AvrPto.     

Pregnancy Rates to Fixed Embryo Transfer of Vitrified IVP Bos indicus,Bos taurus or Bos indicus × Bos taurus Embryos

The pregnancy rates obtained after the transfer of cryopreserved in vitro‐produced (IVP) embryos are usually low and/or inconsistent. The objective of this study was to evaluate the pregnancy rates of Holstein, Gyr and Holstein × Gyr cattle after the transfer of vitrified IVP embryos produced with X‐sorted sperm. Seventy‐two Gyr and 703 Holstein females were subjected to ovum pickup (OPU) sessions, followed by in vitro embryo production using semen from sires of the same breeds. Embryos (1636 Holstein, 241 Gyr and 1515 Holstein × Gyr) were exposed to forskolin for 48 h prior to vitrification. The pregnancy rate achieved with Gyr dam and sire was 46.1%, which was similar (p = 0.11) to that of Holstein dam and Gyr sire (40.3%). Crossing Gyr dams with Holstein sires resulted in a pregnancy rate of 38.9% and did not differ (p = 0.58) from the pregnancy rate obtained with the cross between Holstein dams and Gyr sires. The rate obtained with Holstein dam and sire was 32.5%. The average pregnancy rate was 36.6%, and no difference was found in the proportion of female foetuses (88.8%, in average) among breeds (p > 0.05). In conclusion, transfer of cryopreserved X‐sorted embryos represents an interesting choice for dairy cattle. Despite the small differences between pregnancy rates, we highlight the efficiency of this strategy for all of the racial groups studied.     

Infectivity and persistence of an outbreak strain of Salmonella enterica serotype Typhimurium DT160 for house sparrows (Passer domesticus) in New Zealand

AIM: To examine the infective dose, incubation period and disease progression of an isolate of Salmonella enterica serotype Typhimurium definitive type 160 (DT160) originating from a naturally-infected house sparrow (Passer domesticus) during an outbreak of the disease in New Zealand.

METHODS: Thirty-six house sparrows captured from the wild and free of Salmonella spp were divided into six groups of six birds, housed individually, and inoculated orally with phosphate buffered saline (PBS) or 10¹, 10², 10³, 10⁵, 2 × 10⁸ colony forming units (cfu) of the outbreak strain of S. Typhimurium DT160. The birds were observed for 10 days for clinical signs and/or mortality, and faecal samples were collected to determine excretion of S. Typhimurium. The birds were eutha- nised 11 days post-inoculation (p.i.) and a wide range of tissue samples were collected for histopathological examination, and culture and typing of Salmonella spp. Macro-restriction profiling by pulsed-field gel electrophoresis (PFGE) using XbaI was performed for the epidemiological typing of S. Typhimurium DT160 isolates.

RESULTS: Mortality in house sparrows inoculated with S. Typhimurium DT160 was dose-dependent, and 2/6 birds inoculated with ¹⁰⁵ cfu and all six birds inoculated with 2 × 10⁸ cfu died during the study. Infected sparrows displayed few clinical signs, apart from diarrhoea and/or polyuria, fluffed plumage, and sitting on the floor of the cage. Faecal excretion of DT160 occurred briefly in two birds inoculated with 10² cfu and four birds inoculated with 10³ cfu, on most days in five birds inoculated with 10⁵ cfu, and continuously in six birds inoculated with 2 × 10⁸ cfu. DT160 was isolated from the livers of three birds which received 10³ cfu, five birds dosed with 10⁵ cfu, and all six birds given 2 × 10⁸ cfu. Following necropsy, histopathological lesions similar to those seen in the natural disease were observed in the liver or spleen of three birds which received 10³ cfu, and all birds dosed with ≥10⁵ cfu.

CONCLUSION: The results indicate that an isolate of S. Typhimurium DT60 originating from house sparrows in New Zealand is pathogenic to these birds and that the response is dose- dependent. The persistence and excretion of the pathogen may last for at least 10 days. This confirms that sparrows infected with DT160 could be a source of infection to humans and other in-contact animals.