Colorimetric detection of Ehrlichia canis via nucleic acid hybridization in gold nano-colloids.

Canine monocytic ehrlichiosis (CME) is a major thick-bone disease of dog caused by Ehrlichia canis. Detection of this causal agent outside the laboratory using conventional methods is not effective enough. Thus an assay for E. canis detection based on the p30 outer membrane protein gene was developed. It was based on the p30 gene amplification using loop-mediated isothermal DNA amplification (LAMP). The primer set specific to six areas within the target gene were designed and tested for their sensitivity and specificity. Detection of DNA signals was based on modulation of gold nanoparticles' surface properties and performing DNA/DNA hybridization using an oligonucleotide probe. Presence of target DNA affected the gold colloid nanoparticles in terms of particle aggregation with a plasmonic color change of the gold colloids from ruby red to purple, visible by the naked eye. All the assay steps were completed within 90 min including DNA extraction without relying on standard laboratory facilities. This method was very specific to target bacteria. Its sensitivity with probe hybridization was sufficient to detect 50 copies of target DNA. This method should provide an alternative choice for point of care control and management of the disease.

Simple Screening of Listeria monocytogenes Based on a Fluorescence Assay via a Laminated Lab-On-Paper Chip.

Monitoring food safety is essential for protecting the health and safety of consumers. Conventional methods used are time consuming and laborious, requiring anywhere from three to seven days to obtain results. Thus, better monitoring methods are required. In this study, a laminated lab-on-paper chip was developed, and its use for the screening of ready-to-eat seafood was demonstrated. The assay on a chip was based on loop-mediated isothermal DNA amplification (LAMP) of the hly gene of Listeria monocytogenes and fluorescence signal detection via SYBR GoldTM. Overall assay processes were completed in 4.5 h., (including 3.5 h. incubation for the bacteria enrichment, direct DNA amplification with no DNA extraction, and signal detection), without relying on standard laboratory facilities. Only positive samples induced fluorescence signals on chip upon illumination with UV light (λ = 460). The method has a limit of detection of 100 copies of L. monocytogenes DNA per 50 g of sample. No cross-reactivity was observed in samples contaminated with other bacteria. On-site monitoring of the seafood products using this chip revealed that one of 30 products from low sanitation vendors (3.33%) were contaminated, and these agreed with the results of PCR. The results demonstrated a benefit of this chip assay for practical on-site monitoring.

Effect of temperature and humidity on reproductive performance of crossbred sows in Thailand.

This study investigated the influence of season, temperature, and humidity on the reproductive performance of sows under tropical conditions. Data were collected from 11 sow herds from January 2001 to June 2002. Temperature and humidity were recorded daily for each herd from January 2001 to February 2002. Semen used was collected from boars housed in conventional open-air stables (six herds) or in evaporative cooling stables (five herds). A total of 43,875 farrowing records were included in the statistical analysis. Fourteen-day moving averages of daily maximum temperature and minimum humidity were calculated and merged with each reproductive record. ANOVA was applied to the reproductive records. In addition to the fixed effects included in the statistical models (e.g. system, season, parity, temperature, and humidity), the random effect of herd within system was included. The total number of piglets born was analyzed in relation to the climate at previous weaning (NTB-w), at mating (NTB-m), and at farrowing (NTB-f). The housing system of the boars had no significant effect on any of the reproductive variables analyzed. Season (2-month periods) as well as parity number had a significant effect on all reproductive variables analyzed. Increased length of previous lactation had a significant and favorable effect (P < 0.001) on NTB-w, NTB-m, and weaning-to-first-service interval. There were indications that high temperature and humidity (recorded at the herd level) at previous weaning/mating or at farrowing had negative effects on litter size, but these negative influences were not consistent.

Management and sperm production of boars under differing environmental conditions.

The management of boars to ensure good sperm production under differing environmental conditions is a major concern for pig keepers in both tropical countries and countries where there are extreme environmental changes. Such changes create stress in animals and influence the production of spermatozoa. High temperatures during hot summer months may result in lower feed consumption and create stresses that result in the inhibition of spermatogenesis. Although tropical countries do not have a problem with major variations in day length, this can cause problems such as decreased litter size and infertility in other regions of the world. Evaporative cooling systems built into boar accommodation are often used to reduce fluctuations in both temperature and humidity during the hot and humid months seen in tropical countries. The system has become popular in AI boar studs, where it is reported to reduce stress and improve feed consumption. Other management factors, such as housing comfort, social contact, mating conditions and the frequency of mating, are also very important boar management aids that assist good quality semen production; these will be covered briefly in this review. This review will consider primarily those management factors, for example, the management of temperature and humidity using evaporative cooling systems and other techniques that enable AI boar studs to maximize sperm fertility through adjustments to the environment.

Effect of temperature and humidity on sperm morphology in duroc boars under different housing systems in Thailand.

The aim of this study was to investigate the effect of season, temperature, humidity, age of the boar, and semen collection interval on sperm morphology in Duroc boars in Thailand, kept either in a conventional open air system (CONV) or in an evaporative cooling system (EVAP). In total, 1176 ejaculates from 110 sexually mature boars in six CONV herds and five EVAP herds were morphologically examined during a one-year period. Analysis of variance was applied to the data. Minor differences in the sperm morphology traits analyzed were found between the housing systems. There was a significant seasonal effect (two-month periods) on the percentage of morphologically normal spermatozoa (normal1), morphologically normal spermatozoa including spermatozoa with distal cytoplasmic droplets (normal2), proximal cytoplasmic droplets (prox), and sperm head abnormalities (P