Formation of a Reservoir of Sperm in the Oviduct

A reservoir of sperm in the initial segment of the oviduct has been found in several species of domestic and laboratory mammals. Evidently, the reservoir serves to ensure successful fertilization by providing the appropriate number of sperm in the appropriate physiological state for fertilizing oocytes soon after they enter the oviduct. Recent evidence indicates that sperm are trapped in the reservoir by binding to specific carbohydrate moieties on the surface of the mucosal epithelium of the oviduct. A bovine seminal plasma protein has been identified that associates with sperm and confers on them the capacity to bind to the carbohydrate moiety.     

Specific characters of 16S rRNA gene and 16S–23S rRNA internal transcribed spacer sequences of Xylella fastidiosa pear leaf scorch strains

Pear leaf scorch, the only Xylella fastidiosa-induced disease reported from Taiwan, was found in area where the variety Hengshan (Pyrus pyrifolia) was grown. Strains of pear leaf scorch Xyl. fastidiosa (XF-PLS) shared similarities to strains of other host origins in the requirement of complex medium and the exhibition of rippled cell walls, however, recent serological and molecular biology studies showed difference among them. Five strains of XF-PLS were compared with 20 other strains originally isolated from almond, oleander, pecan, plum, peach, mulberry, grapes, citrus, coffee, and sycamore by sequence analyses of the 16S rRNA gene and 16S–23S rRNA internal transcribed spacer region (ITS). When sequences of 16S rRNA gene based on fragment size of 1,537–1,540 bp were compared, the similarity index among 5 XF-PLS strains was 99.3–99.8%, whereas it was 97.8–98.6% between XF-PLS strains and strains from other hosts. When sequences of 16S–23S rRNA ITS based on fragment size of 510–540 bp were compared, the similarity index among 5 XF-PLS strains was 99.0–100%, whereas it was 80.7–82% between XF-PLS strains and strains from other hosts. Multiple sequence alignments led to the identification of 5 polymorphic nucleotides in the 16S rRNA gene among the 25 Xyl. fastidiosa strains, and there were considerable variations in the nucleotide sequences of 16S–23S rRNA ITS between XF-PLS and the other 20 Xyl. fastidiosa strains. The phylogenetic trees revealed that XF-PLS strains were separated from strains of other hosts. Strains of other hosts were divided into four subgroups: strains from (1) oleander, (2) grape, almond M23 and mulberry, (3) citrus and coffee, and (4) pecan, peach, plum, sycamore and almond M12. Results indicate that XF-PLS strains were not closely related to the above-mentioned strains from other hosts and could possibly belong to a new subspecies of Xyl. fastidiosa.     

Estimating the level of disease risk and biosecurity on commercial poultry farms in New Zealand

ABSTRACT

Aims: To collect baseline data on the contact risk pathways and biosecurity practices of commercial poultry farms in New Zealand, investigate the relationship between the farm-level disease contact risks and biosecurity practices, and identify important poultry health concerns of producers.

Methods: A cross-sectional survey of all registered New Zealand commercial poultry operations was conducted in 2016 collecting information on farm demographics, biosecurity practices, and contact risk pathways. Survey responses were used to generate an unweighted subjective disease risk score based on eight risk criteria and a subjective biosecurity score based on the frequency with which producers reported implementing seven biosecurity measures. Producer opinions towards poultry health issues were also determined.

Results: Responses to the survey response were obtained from 120/414 (29.0%) producers, including 57/157 (36.3%) broiler, 33/169 (19.5%) layer, 24/55 (44%) breeder, and 6/32 (19%) other poultry production types. Median disease risk scores differed between production types (p?<?0.001) and were lowest for breeder enterprises. The greatest risk for layer and broiler enterprises was from the potential movement of employees between sheds, and for breeder enterprises was the on- and off-farm movement of goods and services. Median biosecurity scores also differed between production types (p?<?0.001), and were highest for breeder and broiler enterprises. Across all sectors there was no statistical correlation between biosecurity scores and disease risk scores. Producers showed a high level of concern over effectively managing biosecurity measures.

Conclusions: The uptake of biosecurity measures in the commercial poultry farms surveyed was highly variable, with some having very low scores despite significant potential disease contact risks. This may be related to the low prevalence or absence of many important infectious poultry diseases in New Zealand leading farmers to believe there is a limited need to maintain good biosecurity as well as farmer uncertainty around the efficacy of different biosecurity measures. Further research is needed to understand barriers towards biosecurity adoption including evaluating the cost-effectiveness of biosecurity interventions.     

Insecticidal Activity and HPLC Correlation of Thuringiensin from Fermentation and Two-phase Aqueous Separation Processes

Thuringiensin produced by Bacillus thuringiensis subsp. darmstadiensis was further subjected to a two-phase aqueous separation system. A modified HPLC method and a test for quantitative pathogenicity using the house fly Musca domestica were used for analysis of thuringiensin. Within a realistic range of dosages, more effect was observed in the pupal stage than in the larval stage. The percentage effective control rate (ECR) was calculated by (100-percentage emergence); malformed and non-reproductive adults were considered as emerged. Pupal mortality, pupal weight, and ECR after feeding the three-day-old larvae were the measured response criteria for bioassay. The EC₅₀ of thuringiensin for pupae mortality was 1·64 μg ml^-1 diet, and 0·83 μg ml^-1 for mortality of adults. Insecticidal activity of the broth increased with fermentation time-course from 9th to 21st hour. The bioassay curve constructed with three-hour sampling interval during the fermentation course had good correlation to thuringiensin content as determined by the HPLC method. In the two-phase aqueous separation system, a maximum of 96·7% ECR was achieved with the bottom salt layer, compared to a value of 46·7% with the upper PEG layer. These results suggest that thuringiensin, prepared through a fermentation and recovery process, is suitable for pest control. © 1997 SCI.     

Assessment of potassium speciation in soil using traditional single leaching and modified sequential extraction processes

Purpose

The bioavailability of potassium (K) depends on its speciation distribution in the soil. Different methods are commonly used to estimate K speciation including traditional single leaching (TSL) and sequential extraction process (SEP). However, K speciation is largely affected by soil pretreatment methods. The effects of both TSL and SEP soil pretreatment methods were evaluated.

Materials and methods

The TSL method classifies K speciation content based on bioavailability, while the SEP classifies the metal speciation based on the effects of environmental conditions. These two methods, together with a modified sequential single leaching (SSL) scheme, were used to evaluate five types of soil including soil without potassium fertilization, soil with long-term K fertilization, alkaline soil, red soil, and forest soil. The soil samples were gathered randomly at depths varying up to 30 cm before being dried in air at room temperature. The samples were then ground and mixed before passing through a sieve (10 mesh or 100 mesh) in order to perform K speciation analysis via the modified SSL technique, the TSL method, and the four schemes of SEP.

Results and discussion

Soil pretreatment influenced K speciation, with higher concentration in soil samples sieved through 100 mesh than through 10 mesh. In alkaline soil, potassium was observed to be associated with carbonate. For the various SEP schemes, K speciation was found to be greatest in the residual fraction, with only 3% observed in the carbonate, exchangeable, metal organic complex, or amorphous hydroxides of Fe or Mn. After following the first two steps of the SEP schemes, the available K was similar to that of the TSL method. Distribution of non-exchangeable K using the TSL method was comparable with the five combined SEP extraction steps which were all affected by environmental conditions.

Conclusions

Pretreatment affected K speciation distribution and total amount of metal in the soil. The 100 mesh sieve was more effective in estimating K soil speciation. The SEP method was acceptable for estimating K speciation, with the Krishnamurti et al. (Analyst 120:659–665, 1995) scheme as a useful appraisal of K bioavailability. Combination analyses using both TSL and SEP methods are useful techniques to enable a better understanding of K speciation transformation in soil.

     

Vaginal Neurofibroma in a Hysterectomized Poodle Dog

A 15‐year‐old, spayed, female poodle dog was presented for evaluation of a mass of tissue prolapsed from the vulva. The dog had been hysterectomized when it was 5 years old. A vaginal mass had been removed approximately 10 months before presentation. Haematological and serum biochemistry analyses demonstrated mild leucocytosis and glycaemia. A vaginal smear was predominantly made up of parabasal cells and intermediate cells with no neoplastic cells. Thoracal and abdominal radiographic findings were unremarkable. The ovaries could not be identified using abdominal ultrasonography. A midline exploratory laparotomy identified both ovaries that were surgically excised. The vaginal mass was also removed following an episiotomy procedure. Histopathological examination of the mass demonstrated that it was a neurofibroma. Both ovaries had cystic changes. Four months after the surgery, the owner reported that the dog was clinically normal. To the authors’ knowledge, this is the first reported case of a vaginal neurofibroma after an incomplete ovariohysterectomy in the dog.     

Antcin B and its ester derivative from Antrodia camphorata induce apoptosis in hepatocellular carcinoma cells involves enhancing oxidative stress coincident with activation of intrinsic and extrinsic apoptotic pathway

The triterpenoids methylantcinate B (MAB) and antcin B (AB), isolated from the medicinal mushroom Antrodia camphorata , have been identified as strong cytotoxic agents against various type of cancer cells; however, the mechanisms of MAB- and AB-induced cytotoxicity have not been adequately explored. This study investigated the roles of caspase cascades, reactive oxygen species (ROS), DNA damage, mitochondrial disruption, and Bax and Bcl-2 proteins in MAB- and AB-induced apoptosis of hepatocellular carcinoma (HCC) HepG2 cells. Here, we showed that MAB and AB induced apoptosis in HepG2 cells, as characterized by increased DNA fragmentation, cleavage of PARP, sub-G1 population, chromatin condensation, loss of mitochondrial membrane potential, and release of cytochrome c. Increasing the levels of caspase-2, -3, -8, and -9 activities was involved in MAB- and AB-induced apoptosis, and they could be attenuated by inhibitors of specific caspases, indicating that MAB and AB triggered the caspase-dependent apoptotic pathway. Additionally, the enhanced apoptotic effect correlates with high expression of Fas, Fas ligand, as well as Bax and decreased protein levels of Bcl-(XL) and Bcl-2, suggesting that both the extrinsic and intrinsic apoptosis pathways were involved in the apoptotic processes. Incubation of HepG2 cells with antioxidant enzymes superoxide dismutase and catalase and antioxidants N-acetylcysteine and ascorbic acid attenuated the ROS generation and apoptosis induced by MAB and AB, which indicate that ROS plays a pivotal role in cell death. NADPH oxidase activation was observed in MAB- and AB-stimulated HepG2 cells; however, inhibition of such activation by diphenylamine significantly blocked MAB- and AB-induced ROS production and increased cell viability. Taken together, our results provide the first evidence that triterpenoids MAB and AB induced a NADPH oxidase-provoked oxidative stress and extrinsic and intrinsic apoptosis as a critical mechanism of cause cell death in HCC cells.