Mutation Techniques for Gene Discovery and Crop Improvement

One of the most important breakthroughs in the history of genetics ）was the discovery that mutations can be artificially induced in organisms （van Harten, 1998）. Artificially induced mutations, by physical and chemical mutagens, have greatly advanced the understanding of genetics of higher organisms. Starting in the late 1960＇s, the International Atomic Energy Agency （IAEA） and the Food and Agriculture Organization （FAO） of the United Nations sponsored extensive research on mutation induction and their application to breeding of food and indnstrial crops that resulted in the introduction of new varieties of rice, wheat, barley, apples, citrus, sugar cane, banana, and others （more than 2 500 officially released new varieties are to be found in the FAO/IAEA Mutant Varieties Database） （hitp：//www-mvd.iaea.org/MVD/default.htm）. However, the usefulness of mutation techniques has been underappreciated in research communities, particularly during the last decade, when more and more researchers and breeders were rushing into molecular marker techniques and transgenic plants. In this paper, after a brief review of the past accomplishments of mutation induction and its application, we discuss the uniqueness of induced mutations in gene discovery and how to integrate induced mutants into functional genomics programs;     

Subjective and objective evaluation of veal lean color

Because veal lean color continues to be a primary factor that determines veal carcass value and is typically assessed by subjective means, it is important to explore objective methods for color assessment. Objective and subjective evaluations of veal flank and breast lean color were compared as predictors of longissimus lean color at 24 h postmortem. One hundred fifty special-fed Holstein veal calves were Kosher-slaughtered with blood samples collected upon exsanguination and analyzed for hematocrit and hemoglobin content. Lean color was evaluated in the flank and breast at 0, 6, 12, and 24 h postmortem. Color of the longissimus was evaluated at 6 h, when possible, and at 24 h. A panel of three trained individuals used a 5-point color standard developed in the Netherlands to visually evaluate lean color. A Minolta Chromameter CR-300 was used to obtain L*, a*, and b* values. A plant employee assigned packer grades at slaughter. Temperature and pH were also measured at each time period. Hemoglobin was more highly correlated than hematocrit with colorimeter values. Hemoglobin levels correlated well with a* values of the flank at 0 h postmortem (r = 0.52) although the correlation declined at 24 h (r = 0.30). The correlation between packer grades and 24-h visual loin color was r = 0.41. Visual loin color at 24 h postmortem was selected as the predicted variable for regression analysis. Temperature and pH did not contribute significantly to any prediction equations. The equation using breast L*, a*, and b* values at 24 h postmortem to predict 24-h loin color gave a higher prediction coefficient (R2 = 0.44) than the corresponding equation using 0-h breast values (R2 = 0.28). Objective measurement of lean color may be useful in veal carcass grading because it is more precise than subjective methods and would allow for uniformity among processing plants.     

Long term use of hydraulic artificial urethral sphincters in nine dogs from New Zealand with urethral sphincter mechanism incompetence

AIMS: To report on the long-term outcomes of hydraulic artificial urethral sphincter (HAUS) placement for the correction of urethral sphincter mechanism incompetence (USMI) in New Zealand dogs.

METHODS: Retrospective data were obtained from cases of dogs which had a HAUS placed after failed medical and/or surgical management of USMI between August 2012 and November 2016. Owner assessment of urinary incontinence was evaluated by an online survey in May 2017 using a visual analogue scale (0 being normal, 100 being severely affected) for the frequency, volume and severity of any straining to urinate, immediately prior to the placement of the HAUS and at the time of the survey. The number of days between surgery and the completion of survey were recorded.

RESULTS: Seven females and two male dogs, which were all desexed except for one female, were eligible for inclusion in the study. The period of follow-up following HAUS placement ranged from 206–1,685 days. Following HAUS placement, frequency and volume of urinary incontinence decreased for six dogs and were practically unchanged for three dogs. The median frequency score decreased from 70 to 13 and the volume score decreased from 73 to 12. There was no consistent change in the perceived degree of straining to urinate. Complications occurred in three dogs; one required repositioning of a dislodged injection port, one required management for haematuria and a hypoplastic bladder, and one required surgical removal of fibrous tissue around the HAUS cuff.

CONCLUSIONS AND CLINICAL RELEVENCE: HAUS placement was an effective method for the treatment of persistent USMI in most dogs and provided good clinical results based on owner assessment. The technique was associated with few complications and allowed successful long-term control of urinary incontinence without the need for medical management.     

LOW-FIELD MRI AND ARTHROSCOPY OF MENISCAL LESIONS IN TEN DOGS WITH EXPERIMENTALLY INDUCED CRANIAL CRUCIATE LIGAMENT INSUFFICIENCY

Little is known about the magnetic resonance imaging (MRI) appearance of canine meniscal lesions. The aim of this study is to describe the MR appearance of meniscal lesions in dogs with experimentally induced cranial cruciate ligament (CCL) deficiency. The pilot study revealed dogs weighing approximately 10 kg to be too small for meniscal evaluation on low-field MRI. In the main study, dogs weighing approximately 35 kg were used. The left CCL was transected and low-field MRI was performed regularly until 13 months post-surgery. Normal menisci were defined as grade 0. Intrameniscal lesions not reaching any surface corresponded to grade 1 if focal and to grade 2 if linear or diffuse. Grade 3 lesions consisted in linear tears penetrating a meniscal surface. Grade 4 lesions included complex signal changes or meniscal distortion. Between 2 and 13 months post-surgery, all dogs developed grade 4 lesions in the medial meniscus. Most of them corresponded to longitudinal or bucket handle tears on arthroscopy and necropsy. Two dogs showed grade 3 lesions reaching the tibial surface of the lateral meniscus on MRI but not in arthroscopy. Such tears are difficult to evaluate arthroscopically; MRI provides more accurate information about the tibial meniscal surface. Grades 1 and 2 lesions could not be differentiated from presumably normal menisci with our imaging technique. An MRI grading system better adapted to canine lesions has yet to be developed. MRI is a helpful tool for the diagnosis of complete tears in the canine meniscus, especially in larger dogs.     

Evaluation of an Immunochromatographic Test to the Diagnosis of Canine Brucellosis Caused by Brucella canis

This study evaluated the performance of an immunochromatographic test (ICT) for the diagnosis of canine brucellosis caused by Brucella canis, comparing its results with that of the rapid slide agglutination test with and without the use of 2‐mercaptoethanol and the agar gel immunodiffusion test (AGID). The microbiological culture, PCR and clinical examination were used as reference. According to the results obtained in clinical examination, blood culture, culture of semen and vaginal swab and PCR in blood, semen and vaginal swab, a total of 102 dogs were divided into three groups: B. canis‐infected dogs (Group 1), B. canis‐non‐infected dogs (Group 2) and dogs with suspected brucellosis (Group 3). The diagnostic sensitivity of RSAT, 2ME‐RSAT, AGID and ICT in Group 1 was, respectively, 75%, 37.5%, 27.8% and 89.58%. The diagnostic specificity of RSAT, 2ME‐RSAT, AGID and ICT in Group 2 was, respectively, 91%, 100%, 100%, and 100%. In dogs with suspected brucellosis, 9.67% were RSAT positive, none was positive by 2ME‐RSAT, 3.22% were AGID positive and 6.45% were ICT positive. The main drawback concerning canine brucellosis diagnosis is the lack of a highly sensitive serological assay to be used as a screening test to the rapid identification of infected animals. The ICT showed a high diagnostic specificity and a diagnostic sensitivity value greater than that observed in the RSAT, 2ME‐RSAT and AGID. However, 10.41% of infected dogs had negative results by ICT. These dogs were positive by microbiological culture and/or PCR, indicating active infection and consequently a higher potential of spreading Brucella. Although rapid and simple to perform, the ICT lacked sensitivity to be used as a screening test.     

Soil formation in the quarries for limestone and clay production in the Ukhta region

The soils forming on the overgrowing technogenic dumps of quarries for limestone and clay production were investigated in the northern taiga (the Ukhta region). The soils are formed under sparse herbaceous plant communities. In the soils on calcareous technogenic eluvium and clay dumps, the processes of humus formation and accumulation predominate. In the soils of the clay dumps, the leaching of carbonates is expressed to a greater extent than in the soils of the limestone quarries. The nitrogen content of organic matter is low in the soils on the technogenic lime substrates and very low on the clay ones. Fulvic acids predominate in the humus composition. At the stages of the soil restoration studied, the zonal trend of pedogenesis is manifested only in the humus accumulation. The calcareous technogenic eluvium is found to be more favorable for the development of microbial communities as compared to the clay substrates with their small microbial biomass. However, there are no features pointing to the development of zonal soil profiles. This fact attests that, in the first 20–30 years, the soil-forming potential in the northern taiga is insufficient for the initiation of the zonal processes.