Image analysis as a tool to facilitate selective breeding of quality traits in rainbow trout

A quantitative genetic analysis was performed to assess the suitability of automated image analysis of cutlets as a selection tool to genetically improve flesh composition and colour in large rainbow trout. Fish were reared on two diets with different lipid and protein content to assess the robustness of the image analysis method across different nutritional environments, and the strength of potential genotype-by-diet interactions. Chops were scanned and digitally analysed for colour (chroma, hue, and lightness) and for areas of lipid stripes, dorsal lipid, red muscle and white muscle. Percents of lipid and white muscle area to whole chop area were compared to percent chop lipid and protein recorded by chemical analysis. The results showed that on both diets, percent chop lipid and percent white muscle area displayed high phenotypic (≥ 0.42) and genetic correlations (≥ 0.70) with the respective chemical lipid and protein percents. Moreover, on both diets, the lipid area percents had moderate to high heritabilities (h² = 0.29–0.70) that were of the same magnitude or higher compared to the previously published estimates for chemically analysed lipid traits. These results confirm that regardless of the diet composition, true lipid and protein deposition can be genetically improved by selecting for the respective image analysis traits. Muscle lightness, hue (tint of colour) and chroma (saturation level of colour) displayed moderate to high heritabilities (h² = 0.32–0.46), and phenotypic and genetic correlations between these traits were favourable. Muscle colour can thus be effectively improved by selection. Some genetic constraints for breeding efforts were identified, in terms of lipid deposited at different locations within the body correlating only weakly. Genetic correlations between diets were strongly positive (≥ 0.85), revealing only weak re-ranking of families across the diet treatments.     

Estimation of the carcass composition from a cross-section of the rib-loin from crossbred Japanese Black × Limousin F2 cattle by computer image analysis

The carcass composition of crossbred Japanese Black × Limousin F2 cattle was examined in order to find an accurate carcass composition equation. The test animals included 17 steers and 17 heifers. The 28 image measurements from the area encircling the vertical line to the thoracic vertebra and the line from the thoracic vertebra between the sixth and seventh rib‐bones were measured by computer image analysis. The relationships between the 29 parameters that added the carcass left side weight of the animal and the carcass composition were suggested. The carcass composition included muscle weight, muscle ratio, fat weight and fat ratio. The carcass composition from steers was estimated by an equation composed of these three or four parameters (R² = 90.80%, 79.30%, 90.75% and 73.70%, respectively). The selected parameters were measured without cutting the thoracic vertebra. The carcass composition from heifers was estimated by an equation composed of two to four parameters (R² = 96.15%, 90.98%, 93.60% and 88.22%, respectively). The parameters for the estimation of the muscle and fat weight, and muscle and fat ratio are very similar. Furthermore, the equations using the parameters could estimate the carcass composition from the Japanese Black × Limousin cattle resource population.     

The prevalence of proliferative kidney disease from the kidney and muscle of rainbow and brown trout in Aragón (Spain)

The prevalence and parasite density of PKX (the unknown myxosporean that causes proliferative kidney disease [PKD] of salmonids) were investigated in eight fishfarms in Aragon, Spain. Tissue sections stained with the biotynilated lectin GS-I revealed the presence of this protozoan in only one of the farms. In rainbow trout, the renal prevalence and parasite density peaked in July, but in brown trout the maximum renal prevalence and maximum renal parasite density were reached in May and in July, respectively. In rainbow trout, after the acute phase of the disease, the number of PKX decreases in the kidney but increases in the muscle. In this species of fish, the prevalence and parasite density are much higher in the muscles than in the kidney in October.     

Retracted: Reliable collection of Caspian brown trout (Salmo trutta caspius) sperm using a catheter

The traditional stripping procedure for collecting fish semen is associated with the risk of urine contamination, which may significantly affect semen quality and quantity. The use of a catheter as an alternative method for semen collection may overcome this problem. Therefore, this study compared Caspian brown trout (Salmo trutta caspius) semen parameters (i.e. sperm density, seminal plasma osmolality, motility parameters of spermatozoa analysed using computer‐assisted sperm analysis and fertility) between the traditional stripping method and the use of a catheter. All parameter values of the semen collected with a catheter were significantly higher (p < .05; density = 7.67 ± 1.02 × 10⁹ ml^?1 and osmolality = 279.28 ± 32.84 mOsm kg^?1) than those collected with stripping method (density = 4.85 ± 0.47 × 10⁹ ml^?1 and osmolality = 216.42 ± 20.75 mOsm kg^?1). Semen collected with a catheter was characterized by higher spermatozoa motility compared with sperm collected via stripping. Similarly, the fertilization ability of sperm collected with a catheter was significantly greater (p < .05) than sperm collected with the traditional stripping method. In conclusion, collection of sperm with a catheter was shown to effectively reduce urine contamination and is therefore recommended for the collection of Caspian brown trout sperm.     

Single-strand conformation polymorphism (SSCP) analysis as a new diagnostic tool to distinguish dorsal-spined larvae of the Elaphostrongylinae (Nematoda: Protostrongylidae) from cervids

Single-strand conformation polymorphism (SSCP) was used to genetically differentiate morphologically indistinguishable first-stage larvae (L(1)) of the six species of elaphostrongyline nematodes. A partial fragment (317-336bp) of the first internal transcribed spacer (pITS-1) plus 5' flanking region (76bp of the 18S gene) of the nuclear ribosomal DNA (rDNA) was amplified from individual L(1) of known identity and subjected to SSCP. The results showed that the four species of elaphostrongylines found in North American cervids, Parelaphostrongylus tenuis, P. andersoni, P. odocoilei and Elaphostrongylus rangiferi, could be distinguished from one another based on their distinct (i.e. species-specific) SSCP profiles. In addition, E. alces, a species that occurs in moose in Fennoscandinavia, also had a distinct SSCP profile with respect to the other species of elaphostrongylines. However, the SSCP profiles of E. cervi could not be distinguished from those of E. rangiferi because of a lack of interspecific sequence differences in this region of the ITS-1. The distinct SSCP profiles for the other species were consistent with the interspecific differences in ITS-1 sequences, which ranged from 2 (between P. tenuis and P. andersoni) to 59bp (between genera). The pITS-1 SSCP approach was also used to identify unknown elaphostrongyline L(1) from different hosts and localities in North America. The ability to distinguish between L(1) of the four elaphostrongyline species that occur in North American cervids has important diagnostic and epidemiological implications.     

Nonisotopic detection of Loma salmonae (microspora) in rainbow trout (Oncorhynchus mykiss) gills by in situ hybridization

Loma salmonae, a microsporidian parasite of salmonids of the genus Oncorhynchus, is a significant cause of economic loss in pen-reared chinook salmon (O. tschawytscha). Final stages of L. salmonae infections are easily recognized by the xenomas that form in the gills during sporogony. However, early prexenoma stages of infection (3 weeks or less after infection) are difficult to detect on histologic slides. An L. salmonae-specific single-stranded DNA probe labeled with digoxigenin was used to detect these prexenoma stages of L salmonae by in situ hybridization in experimentally infected rainbow trout. This method allows detection of the parasite in the gills only 2 weeks after infection, providing a sensitive and specific way of detecting L. salmonae during the early stages of infection.     

Genetic structure in Atlantic brown trout (Salmo trutta L.) populations in the Iberian peninsula: evidence from mitochondrial and nuclear DNA analysis

Brown trout (Salmo trutta L.) was sampled in rivers belonging to three different Spanish basins in order to analyse the distribution of genetic variability. The genetic analysis was performed by using two systems and techniques: nuclear DNA was screened through random amplified polymorphic DNAs (screening 2 × 10⁵ bp of the whole genome), and mitochondrial DNA (mtDNA) through sequencing of the hypervariable control region. Genetic distances between the populations were similar using either analysis although some differences arise. For example, some populations of the Tajo basin were very close through nuclear analysis but more distant using mtDNA. Differences between the two DNA sources could be the result of a different evolutionary rate, and the fact that mtDNA is maternally transmitted and differences in sex migration rates will influence the patterns of genetic variation between the transmitted DNAs. Total variation was partitioned using amova showing a clear subdivision among basins although intrapopulation variation remained as high as 62%. A correspondence analysis defined the differences in a three‐dimensional way, clustering the populations according to their common basin. When mtDNA was sequenced, higher variability was noted in the segment between 400 and 600bp of the whole D‐loop sequence, suggesting that these 200bp improved the analysis of the variability more than sequencing the t‐RNA ends of the control region. A comparison was made between the t‐RNA^Pro ends of the 10 populations screened here and the rest of the published sequences found in the literature, leading to a concentration of these populations in group IV which includes all trouts which originate in the Atlantic. The analyses performed suggest that a high genetic variability is present in all populations and that although there has been a probable interference from stocked strains introduced to increase population density, this was only detectable through the variance between rivers which reflect different policies according to the region where the basin is located. However, the genetic analysis using the two approaches allows the control of the natural populations avoiding a loss of their genetic potential.     

Genetic structure in Atlantic brown trout (Salmo trutta L.) populations in the Iberian peninsula: evidence from mitochondrial and nuclear DNA analysis

Brown trout (Salmo trutta L.) was sampled in rivers belonging to three different Spanish basins in order to analyse the distribution of genetic variability. The genetic analysis was performed by using two systems and techniques: nuclear DNA was screened through random amplified polymorphic DNAs (screening 2 × 10⁵ bp of the whole genome), and mitochondrial DNA (mtDNA) through sequencing of the hypervariable control region. Genetic distances between the populations were similar using either analysis although some differences arise. For example, some populations of the Tajo basin were very close through nuclear analysis but more distant using mtDNA. Differences between the two DNA sources could be the result of a different evolutionary rate, and the fact that mtDNA is maternally transmitted and differences in sex migration rates will influence the patterns of genetic variation between the transmitted DNAs. Total variation was partitioned using amova showing a clear subdivision among basins although intrapopulation variation remained as high as 62%. A correspondence analysis defined the differences in a three‐dimensional way, clustering the populations according to their common basin. When mtDNA was sequenced, higher variability was noted in the segment between 400 and 600 bp of the whole D‐loop sequence, suggesting that these 200 bp improved the analysis of the variability more than sequencing the t‐RNA ends of the control region. A comparison was made between the t‐RNA^Pro ends of the 10 populations screened here and the rest of the published sequences found in the literature, leading to a concentration of these populations in group IV which includes all trouts which originate in the Atlantic. The analyses performed suggest that a high genetic variability is present in all populations and that although there has been a probable interference from stocked strains introduced to increase population density, this was only detectable through the variance between rivers which reflect different policies according to the region where the basin is located. However, the genetic analysis using the two approaches allows the control of the natural populations avoiding a loss of their genetic potential.     

Primordial germ cells (PGCs) as a tool for creating transgenic chickens

The transgenic chicken has great potential as a bioreactor for the production of valuable pharmaceutical proteins, notably in the oviduct/egg. Whereas conventional transgenic approaches have significant limitations in this species, an alternative approach employing primordial germ cells (PGCs), the progenitor cells to ova and spermatozoa, has now been successfully applied to the insertion of exogenous genes into birds. Recent developments in manipulating avian embryos make it possible to produce germline chimeras derived from transferred PGCs. In this review we describe the migration pathway of chicken PGCs during early development. We then summarize different methods for the isolation of PGCs and the diversity of techniques used to introduce genes into these cells. Finally, we describe an in vitro assay for testing tissue-specific vectors designed to express heterologous proteins in transgenic chickens.     

Ultrastructure of the Hinny (Equus asinus × Equus caballus) Seminiferous Epithelium

The gonads and the germinative cells of 3 male hinnies were studied with light and transmission electron microscopy with the aim to observe the development of germ cells and verify the morphological modifications due to the hybridization. The hinny seminiferous epithelium presented Sertoli cells and spermatogonia with normal features and anomalous spermatocytes I. The other cells from the spermatogenic sequence were not seen. Most of the alterations began to occur in the cytes I, which presented nuclear vacuolization and deposits of amorphous material between the carioteca and the nuclear lamina, forming vesicles, or exaggerated chromatin condensation, resulting in pyknosis. In the cytoplasm vacuolization was also observed, besides organelle destruction.
The arrest of meiosis due to lock of chromosome homologies leads to germinative cell degeneration and, therefore, the spermatogenesis arrest. This fact causes a profound alteration in the seminiferous epithelium morphology in comparison with the parental species.     

Supplementary effect of peppermint (Mentha × piperita) on dry matter intake, digestibility, ruminal fermentation and milk production in early lactating dairy cows

The aim of this research was to investigate the effect of peppermint feeding on dry matter intake, nutrient digestibility, ruminal fermentation and milk production in early lactating cows. Four Holstein cows were offered a diet with 5% dried peppermint and four Holstein cows remained on a diet without 5% of dried peppermint on a dry matter basis. The addition of peppermint to feed did not affect dry matter intake, although the eating time of feed was increased by mixing the feed with peppermint. There were no significant differences in the nutrient digestibilities between the two treatments. The ruminal ammonia and volatile fatty acids concentrations were similar in the two treatments, however, peppermint ingestion by cows led to a decrease in ruminal pH. The lowered pH value was within the stable pH condition range. No significant differences in the treatments were observed in milk production or milk composition except for the milk fat content. These results suggest that feeding peppermint to early lactating cows had little effect on their dry matter intake, nutrient digestibility, ruminal fermentation and milk production.     

新疆家蚕种质资源RAPD多态性研究

以家蚕卵为材料。选用重复性较好的12种随机引物，对15份新疆家蚕种质资源材料进行了DNA多态性扩增(RAPD)研究，研究结果表明，新疆家蚕种质资源材料的DNA多态性较为丰富，共获得153个RAPD标记，系统聚类分析表明新疆家蚕种质资源材料的亲缘关系较近、在相似系数0．8的水平下聚为一类，遗传距离最大为0．3985，最小为0．1296。     

Remote monitoring system as a tool for calving management in Mediterranean Buffalo heifers (Bubalus bubalis)

Buffalo breeding is common in Southern Italy. Dystocia compromises dam's and newborn health and welfare. Difficult parturition could be solved through prompt calving assistance, even if the identification of the beginning of delivery is challenging. Herein, we aimed to evaluate a remote calving alarm system in 15 Mediterranean buffalo heifers. An intravaginal probe was placed close to the external cervical os once premonitory signs of delivery were observed. No vaginal discharge nor signs of discomfort were notified in the days following the insertion of the probe. Heifers calved from 48 to 72 hr after the alarm was activated. The system correctly warned the farm personnel at the beginning of stage II of parturition, except for 2 cases. In the former, the intravaginal probe was expelled but the poor carrier network coverage negatively affected phone's signal quality; in the latter, recurrent vaginal prolapse was responsible for non-retention of the probe. Overall median expulsive phase was 68 ± 8 min, while the expulsion of a female calf took 54 ± 22.0 min and 90 ± 34.0 min in males, with significant difference (p =.02). Deliveries were homogeneously distributed across a 24-hr interval. No retention of foetal membranes nor metritis was identified at postpartum clinical examination. The calving alarm system used in this work was well tolerated in buffalo heifers. The introduction of smart technology in buffalo farming could contribute to the overall farm net return by reducing calf losses, especially for calves born from sexed-sorted semen, and by increasing animal welfare through quick resolution of dystocia. Further studies will be necessary to evaluate the net return in buffalo farms which will implement a remote calving alarm system on a wider population.     

Randomly amplified polymorphic DNA (RAPD) analysis of Mycoplasma gallisepticum isolates from turkeys from the central valley of California.

Randomly amplified polymorphic DNA (RAPD) analysis was used to investigate the molecular epidemiology of 26 Mycoplasma gallisepticum (MG) isolates obtained from turkeys located in the central valley of California. The MG isolates were recovered from 5 different companies and 13 ranches. Each company had unique MG strains. No evidence of spread of MG between companies was detected. RAPD analysis of MG isolates within a ranch during an outbreak revealed only a single strain involved in each outbreak. RAPD analysis identified an isolate from 1 ranch with a banding pattern identical to that of the 6/85 vaccine strain, which had been used on that particular ranch. Similar RAPD banding patterns of isolates from different ranches within the same company suggested horizontal spread of MG between ranches. The use of 2 primer sets in RAPD analysis was critical to prevent misinterpretation of relationships between different isolates.     

Functional proteomic analysis of crossbred (Holstein Friesian × Sahiwal) bull spermatozoa

Male infertility is one of the prime concerns of dairy cattle production. The study was designed to find out differentially expressed proteins in categorized crossbred (Holstein Friesian × Sahiwal) bull semen to serve as potential biomarkers for male infertility. Frozen crossbred bull semen with satisfactory phenotypic records were defined as “good” and “poor” based on their fertility rates. A total of 1,547 proteins were detected in bull spermatozoa using liquid chromatography–mass spectrometer (LC‐MS/MS) analysis. Results revealed that 558 (36.1%) and 653 (42.2%) proteins were expressed to good and poor quality bull spermatozoa, respectively. A total of 336 proteins (21.7%) were reported to be unique for both good and poor quality bull semen, and among the common proteins, 224 (66.7%) and 112 (33.3%) were up‐ and downregulated in good and poor quality categorized bull semen, respectively. Gene Ontology analysis of global proteomes identified different signalling pathways, and most of them were related to cellular motility, immune systems as well as cellular metabolisms. The distinctive presence of some of the proteins may provide an insight into the molecular mechanistic role played by these proteins in crossbred bull infertility.     

Post mortem computed tomography as a complementary tool for diagnosing cholangiohepatitis in a giant panda (Ailuropoda melanoleuca)

A geriatric female giant panda developed grave signs of illness and was diagnosed with suspected hepatobiliary tract obstruction or other severe hepatic disease such as advanced cholangiohepatitis. The giant panda was euthanized and post mortem computed tomography was performed prior to necropsy. Common bile duct obstruction at the major duodenal papilla by a mineral attenuating calculus causing dilatation of common bile and gallbladder with concurrent multiple areas of liver abscess were detected by postmortem computed tomography. These were confirmed with gross necropsy. This is the first case report of common bile duct obstruction by mineral calculus with concurrent severe cholangiohepatitis in a giant panda.     

Comparison of the susceptibility of brown trout (Salmo trutta) and four rainbow trout (Oncorhynchus mykiss) strains to the myxozoan Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD)

Differences in susceptibility to the myxozoan parasite Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), between four strains of rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta) were evaluated. Fish were exposed to water enzootic for the parasite in the field for 5 days and were subsequently transferred to the laboratory. Relative parasite load was determined after 2, 3 and 4 weeks post-exposure (wpe) by quantitative real-time PCR (qPCR) of kidney samples and number of parasite stages was determined in immunohistochemical stained sections of kidney, liver and spleen tissues. According to qPCR results, the highest amount of parasite DNA per equal amount of host tissue at all time points was measured in brown trout. Two of the rainbow trout strains showed lower relative parasite load than all other groups at the beginning of the experiment, but the parasite multiplied faster in these strains resulting in an equal level of relative parasite load for all rainbow trout strains at 4 wpe. A weak negative correlation of fish size and parasite load was detected. Only in samples of a few fish, single stages of T. bryosalmonae were found in sections stained by immunohistochemistry impeding quantitative evaluation of parasite numbers by this method. The results indicate a differential resistance to T. bryosalmonae between the rainbow trout strains investigated and between rainbow trout and brown trout.