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Role of male-female interaction in regulating reproduction in sheep and goats.

The induction of synchronous ovulatory activity in anovulatory sheep and goats after the introduction of males, the 'male effect', has probably been used to advantage since these species were domesticated and the underlying physiological and behavioural mechanisms have been progressively elucidated over the past 50 years. Less well understood is the analogous effect of oestrous females on males. This review examines the nature and importance of these male-female interactions in sheep and goats, and describes the most important internal and external factors influencing the reproductive outcomes of such interactions. It is proposed that the male and female effects are both components of a self-reinforcing cycle of stimulation that, under ideal conditions, culminates in the synchronous very rapid onset (within days) of fertile reproductive activity. However, precisely because of the speed of this response, it is suggested that mechanisms have evolved to limit its efficacy, and thus prevent conception at inappropriate times. The complexity of these factors and the interactions between them are highlighted, and a broad conceptual framework for understanding them is proposed based upon an appreciation of variation in both the responsiveness of the target animal and the quality of the signal from the signalling animal.

Animal Nutritional Physiological Phenomena↗

Genetic diagnosis of claudin-16 deficiency and sex determination in bovine preimplantation embryos.

Renal tubular dysplasia is an autosomal recessively inherited disorder in Japanese black cattle that is due to deletion mutations in the claudin-16 gene and causes chronic renal failure and death of affected animals. Here, we report a multiplex-PCR procedure to determine the genotype for claudin-16 deficiency in preimplantation embryos. The presence or absence of the wild-type and mutant allele(s) was precisely detected with the multiplex-PCR using as little as 5 pg of genomic DNA from leukocytes. When biopsied embryo cells were examined for claudin-16 deficiency, 97.2% of genotypes were consistent with the PCR results obtained for the corresponding embryos. In addition, sexing of embryos by PCR was performed using an aliquot of DNA extracted from biopsied embryo cells, and determination of claudin-16 genotype and sex was successfully achieved with an efficiency of 91.7% for claudin-16 genotyping and 83.3% for sexing. The production of a 100-day fetus that was male and homozygous for claudin-16 deficiency, as expected from the analysis of biopsied embryo cells, gave evidence of the reliability and applicability of this procedure for preventing the transmission of this disease and for enabling advances in animal breeding.

Alleles↗

The effect of a low energy diet on the concentration of DDT in the adipose tissue of turkeys.

Turkey hens which had completed a breeding season and contained DDT in their fat were divided into 2 groups of 20 hens each. One group was fed a high wheat control diet throughout a 6 week period while another group was fed a low energy diet for 3 weeks and then the control diet for 3 weeks. Biopsy samples of adipose tissue taken initially and at 3 weeks and carcass adipose tissue samples of 6 weeks were analyzed for DDT and DDE. Total DDT concentration in adipose tissue increased when the hens were fed the low energy diet but decreased again when fed the control diet so that the overall change in DDT concentration over the 6 week period was not different for the two groups. The biopsy technique used in this study was successful in greatly reducing variability and improving precision.

Adipose Tissue↗

Leprosy, an overview.

Leprosy is both a public health problem and a model for host-parasite relationship. There is much progress in many areas, and in most, hurdles are not only scientific but also social and financial. The following are goals to reach: 1) progress of growth of M. leprae in animals (to be able to breed armadillos in captivity or to substitute them with other animals); 2) to develop methods for in vitro culture of M. leprae; 3) to obtain antigens from M. leprae and characterize them immunologically; 4) to achieve techniques for objective diagnosis of infection with M. leprae and of leprosy (these techniques will most likely be serologic); 5) to develop methods to direct and manipulate immune response. This should include a better definition of immunologically active cells, their characterization by reliable methods, and an understanding of their functions. Immune control mechanisms and ways to channel them should also be understood; and 6) to have better methods of treatment including flexible adaptable schedules. It will be clear that "solving" the problem of leprosy requires "solving" the social problems in developing countries, as well as unraveling the mystery of the precise control of immune response. As I pointed out, the problems that we face are basically similar to those of autoimmune disorders and of cancer immunology. Research in autoimmune diseases or cancer might provide answers to questions posed by leprosy. It may also be that research in leprosy could supply answers to questions concerning cancer and autoimmunity.

Animals↗

High resolution DNA flow cytometry of boar sperm cells in identification of boars carrying cytogenetic aberrations.

The cytogenetic quality of boars used for breeding determines the litter outcome and thus has large economical consequences. Traditionally, quality controls based on the examination of simple karyograms are time consuming and sometimes give uncertain results. As an alternative, the use of high-resolution DNA flow cytometry on DAPI-stained sperm cell nuclei (CV </= 1.3%) was investigated as a screening method for the cytogenetic quality of boars. By analyzing a series of 25 animals judged normal by their fertility statistics and a series of seven animals with known reciprocal translocations, a model for identifying sperm cells from cytogenetically aberrant animals was proposed. This model was applied to a series of 50 uncharacterized animals. The model successfully identified a mosaic or chimaeric carrier of an aberrant X chromosome. However, implementation of this technique for screening purposes would necessitate essential improvements in standardization and measurement precision.

Animals↗

Immunohistochemical and histopathologic features of 14 malignant fibrous histiocytomas from Flat-Coated Retrievers.

Flat-Coated Retrievers seem to be at increased risk of developing soft-tissue sarcomas, and undifferentiated round cell or spindle cell sarcomas account for approximately 59% of sarcomas in the breed. In an attempt to classify these tumors further, formalin-fixed, paraffin-embedded sections from 14 undifferentiated sarcomas from Flat-Coated Retrievers were reviewed and examined with a panel of histologic and immunohistochemical stains. The panel included vimentin, desmin, Myo D1, smooth muscle actin, cytokeratin, S100, von Willebrand factor (factor VIII), Mac 387, CD3, major histocompatibility complex (MHC) class II, and CD79a. The majority of the sarcomas showed greater than 70% staining for MHC class II. We conclude that these undifferentiated sarcomas in Flat-Coated Retrievers belong to a spectrum of tumors with varying proportions of characteristic cell types and morphologic features, some of which fit the diagnostic criteria for malignant fibrous histiocytoma. Many of these sarcomas seem to have a significant myofibroblast component and a mild or moderate T cell infiltrate but the precise cell lineage is still uncertain.

Animals↗

[The dose precision of PMSG in young and old sows in the process of biotechnical ovulation synchronization. 1. Comparison of diagnostic laparotomy].

Treatment was applied, on two farms, to 272 gilts and 169 adult sows, 24 hours after discontinuation of Suisynchron application to the gilts or weaning of piglets of adult sows. Included were 600, 800 or 1,000 IU of PMSG for the gilts and 750, 1,000 oder 1,250 IU of the same batch (Pregmagon-Dessau) for the adult sows for ovarian stimulation. This was followed by application of 500 IU of HCG to the gilts or 300 micrograms Gn-RH vet. "Berlin-Chemie" + 300 IU HCG to the adult sows for synchronised ovulation. Laparotomy was performed on the gilts on the fifth day from PMSG application and on the adult sows on the fourth day. Average ovariotropic stimulation was "mild" in response to lower doses (14.8 follicles in gilts and 20.4 follicles in adult sows) but was "pronounced" in response to medium doses (19.4 or 22.7 follicles, respectively). The pharmacological concept of "mild" exogenic ovarian stimulation by low dosage at the threshold of effectiveness should deserve more attention, last but not least, for breeding aspects. The two groups of animals on both farms differed considerably from one another with regard to reactivity, which cannot be etiologically interpreted for the time being.

Animals↗

Energy needs of companion animals: matching food intakes to requirements throughout the life cycle.

An animal's primary demand from its diet is for energy. Unlike an individual nutrient, there is no range of acceptable intakes, but rather a specific energy requirement for a given set of conditions. Accurate assessments of both food energy density and the energy needs of the animal are implicit in providing reliable feeding recommendations. The metabolizable energy content of a food is best measured using in vivo testing in the target species. However, the demands of biological testing mean that a set of factors is desirable to provide reasonable estimates. Assessment of energy requirement in dogs is confounded by their wide range of body weights. It should be expressed on an allometric basis, but the precise value for the exponent is still a matter of some debate. For cats recent studies suggest there may be an allometric relationship over a weight range of 2.5-6.5 kg. Energy requirement is dependent on a number of other factors, in particular, activity, environmental temperature and life stage. Demand increases most in lactation (four times) and growth (up to three times). Old animals show a decline in energy requirement, but this may reflect principally a decrease in activity, rather than any underlying change in metabolism.

Animals↗

The hepatocytes of the brown trout (Salmo trutta f. fario): a quantitative study using design-based stereology.

A stereological study was performed on brown trout hepatocytes aiming to disclose whether there are basic gender differences when minimal levels of sex hormones exist, and also to establish a platform for both interspecific comparisons and physiological correlations. We used the so-called "design-based stereology" (with no shape, size or orientation assumptions) and also some new related statistics. Two-year-old brown trout were collected in April, and the livers were fixed by perfusion. From liver slicing to microscopical field selection, systematic sampling was used. Stereology was applied at light and electron microscopy. Target parameters were the relative and total hepatocyte number, the mean individual hepatocyte volume and surface, and also both relative and total volumes, and surfaces, either of organelles or of cell compartments. Observed variability was usually high, but the precision of estimates was proved to be globally adequate facing the true biological variation amongst specimens. Females had more hepatocytes per liver (1.79x10(9) vs. 1.12x10(9)). Considering the individual hepatocytes, whereas no gender differences were detected in the cell volume, males had higher values of nuclear volume (199 vs. 151 microm3) and surface (170 vs. 131 microm2), endoplasmic reticulum volume (1,300 vs. 824 microm3), and microvilli volume (82 vs. 54 microm3) and surface (1,445 vs. 975 microm2). However, when dealing with quantities per liver, gender differences were found only in the volumes of dense bodies (56 vs. 97 mm3) and of residual cytoplasm (169 vs. 341 mm3)--both volumes were higher in females. Functional implications of data are discussed, namely that females seem to have basic structural traits for coping with the later demands of breeding. Data also support that structural remodelling of hepatocytes occurs after breeding, urging to pursue seasonal studies (namely on lysosomes). We advanced the hypothesis that genders differ in microvilli surface just to maintain an optimal physiological surface-to-volume ratio. Interspecific similarities and differences were disclosed. For example, the number of hepatocytes/cm3 of parenchyma of brown trout was much lower than those reported in rainbow trout, but in both trouts females seem to have an higher cell number. In addition, when comparing the size of hepatocytes of brown trout with that from other fish and mammals it was suggested that major interspecific differences exist.

Animals↗

Development of a new radioimmunoassay for erythropoietin using recombinant erythropoietin.

The development of a 24 hour radioimmunoassay for erythropoietin (EPO) using EPO derived from recombinant DNA as both immunogen and ligand is described in the present paper. Mixed breed rabbits immunized with 10 micrograms/kg of EPO derived from a stably transfected cell line (Elanex Pharmaceuticals Inc., Bothel, Washington, USA, through McDonnell Douglas Corp., St. Louis, Missouri, USA; "MD") produced antibodies to EPO with high titer (up to 1:896,000 final dilution in the tube), high affinity (8.4 x 10(11) liter/M), and good specificity. Purified EPO from the above source or from AmGen Biologicals (Thousand Oaks, California, USA; "AG") were successfully radioiodinated with the chloramine-T method and used as ligand in the radioimmunoassay. Standard dose-response curves prepared with EPO from both commercial sources were not significantly different and showed a sensitivity of 0.75 to 0.96 mU/tube. The dose-response curves in both systems also showed parallelism with serially diluted serum from a patient with aplastic anemia. Within-assay and between-assay precision were determined by assaying multiple replicates of a serum pool. Recovery of exogenous EPO added to a serum pool averaged 97% for both systems. The range of normal human serum EPO was determined by assaying the sera of 153 hematologically-normal adult subjects and was found to be 1.1 to 27.3 mU/ml for MD EPO and 0.5 to 16.7 mU/ml for AG EPO. Sera from several patients with hematologic abnormalities were also assayed, including those of 36 patients with anemia of end-stage renal disease (mean +/- SEM, 29.5 +/- 4.0 mU/ml; P less than 0.01). In conclusion, this new, more rapid and sensitive radioimmunoassay system can be used to measure EPO levels in sera from normal human subjects and patients with several types of anemia, and should also be very useful in therapeutic drug monitoring of patients receiving EPO from various commercial sources.

Anemia↗

ClearDepthIAS enables automated high-throughput quantification of roots in soil-grown taproot crops.

Understanding root system architecture is critical for improving crop productivity and resilience, yet phenotyping root traits such as root growth angle and rooting depth remains technically challenging, especially at high throughput. Here, we present ClearDepthIAS, a high-throughput imaging and analysis platform that enables nondestructive, automated quantification of root architecture traits in taproot system crops. By capturing and stitching 360&#xb0; images of roots growing along the transparent walls of pots and applying deep learning-based segmentation (ClearDepth-WRT), we measured wall root shallowness (WRS)-a proxy for root growth angle-with high precision. We demonstrated for the tap root systems of soybean and canola that the system accurately detects root tips, quantifies their vertical distribution, and extracts biologically meaningful traits such as root area, distribution indices, and growth angles. Validation experiments in canola and soybean demonstrated that WRS can correlate with root crown architecture in mature plants, both in greenhouse and field settings. Furthermore, WRS and root distribution indices derived from ClearDepthIAS are predictors of early root architecture and can be correlated with root biomass distribution across soil depths under field conditions; however, environmental interactions may influence these relationships and weaken or even negate such correlations, as observed when comparing field to field variation in root system architecture. Our system enables efficient phenotyping of genetically diverse populations, with medium to high trait heritability, supporting its utility for genome-wide association studies and breeding. ClearDepthIAS accelerates the development of root ideotypes for improved resource acquisition and carbon sequestration, offering a scalable tool for supporting climate-resilient agriculture.

Plant Roots↗

Polymorphic AP-1 binding site in bovine CSN1S1 shows quantitative differences in protein binding associated with milk protein expression.

Polymorphisms in 5'-flanking regions of milk protein encoding genes can influence the binding activity of the affected response elements and thus have an impact on the expression of the gene products. However, precise quantitative data concerning the binding properties of such variable response elements have so far not been described. In this study we present the results of a quantitative fluorescent electromobility shift assay comparing the allelic variants of a polymorphic activator protein-1 binding site in the promoter region of the bovine alphas1-casein encoding gene (CSN1S1), which is affected by an A-->G exchange at -175 bp (CSN1S1(-175bp)). A supershift assay using a commercial c-jun antibody was carried out to verify the specificity of protein binding. The gel shift analysis revealed specific and significantly reduced protein binding of oligonucleotides containing the G variant of the CSN1S1(-175bp) binding site. Further investigations comprised genotyping of the variable CSN1S1(-175bp) activator protein-1 element by an NmuCl restriction fragment length polymorphism in 62 cows of the breed Simmental and 80 cows of the breed German Holstein. Single milk proteins from at least 4 milk samples per cow were quantified by alkaline urea polyacrylamide gel electrophoresis. Homozygotes for CSN1S1(-175bp)*G were not observed, and the allele frequencies were 0.19 in Simmental and 0.05 in German Holstein. Carriers of CSN1S1(-175bp)*G showed higher content (%) as well as quantity (g/d) of alphas1-casein than CSN1S1(-175bp)*A homozygotes, independent of breed. We assume that the positive association of the CSN1S1(-175bp)*G variant with CSN1S1 expression is likely to be caused by a reduced affinity of the affected response element to a c-jun-containing CSN1S1 dimer with repressor properties.

Alleles↗

[Optimized methods for measuring coagulation factors II, IV, VII and X in cats].

Since the single factor activity of the cat differed in part distinctly from humans (factor II: 94%, factor V: 500%, factor VII: 125%, factor X: 62%) the method had to be optimized, above all to exclude an influence of a reactive increased factor V-activity on the results of the other coagulation factors. Dilution series of cat pool plasma (CPP) (n = 50 cats) showed, that a commercial human placenta thromboplastin was more suitable as activating reagent than rabbit brain thromboplastin. Using the required sample predilution (1:40), measurements with the latter reagent were more precise when measured on the relation between the imprecision and the time interval between different CPP-dilution steps. Reference curves were prepared for CPP using the optimized method and human placenta thromboplastin (50 microliters 1:40 prediluted sample and 50 microliters deficient plasma were incubated at 37 degrees C for 1 minute, addition of 200 microliters Ca-thromboplastin) and reference ranges were calculated using the values of 57 cats (factor II: 74-126%, factor V: 42-184%, factor VII 56-150%, factor X: 65-143%).

Animals↗

Vitamin E content of foods: comparison of results obtained from food composition tables and HPLC analysis.

BACKGROUND & AIMS: Data on vitamin E content of foods are essential for nutrition research and its application. The aim of this study was to investigate the precision of calculated vitamin E content of prepared meals. METHODS: The vitamin E content of 69 dishes of a menu cycle sampled at two occasions were calculated using 4 different food composition tables (FCT) and measured by HPLC. RESULTS: Data were complete for 50-69 dishes. The proportion of dishes with differences between FCTs < or = 20% were 17-100%. Differences between HPLC and the Souci-Fachmann-Kraut table were < or = 20% in 8/46 dishes for alpha- and in 14/46 dishes for gamma-tocopherol. Differences fell in the order of baked > raw > fried/roasted > boiled > mixed prepared foods. The 2 samplings taken 6 months apart showed considerable differences. CONCLUSIONS: There are substantial differences in calculated/measured vitamin E content of prepared foods: (i) between different FCTs; (ii) between FCTs and HPLC, and (iii) between different seasons. This can be explained by intrinsic variability (breeding, season, country of origin, ripeness, freshness) and food processing, as well as selection of FCTs and should be taken into account when interpreting data of dietary intervention studies.

Chromatography, High Pressure Liquid↗

From dysbiosis to resilience: Microbiome engineering for sustainable shrimp aquaculture.

The intensification of shrimp aquaculture has increased exposure to disease, environmental perturbations, and antimicrobial pressure, making microbial stability increasingly relevant to sustainable production. Microbiome stability-encompassing resistance to disturbance and resilience of functional recovery-provides an ecological framework for understanding how shrimp and culture-environment microbial communities respond to intensive farming. This review examines the transition from microbial homeostasis to dysbiosis and evaluates how microbiome engineering could redirect disrupted communities towards resilient states. Evidence is integrated across the intestine, hepatopancreas, rearing water, sediment and biofloc to assess how host genetics, ontogeny, diet, culture conditions, antibiotics and pollutants shape microbiome assembly and destabilization. Disease-associated changes in acute hepatopancreatic necrosis disease, white faeces syndrome, Enterocytozoon hepatopenaei infection, and white spot syndrome virus infection are critically evaluated, with explicit separation of associations, pathogen-induced dysbiosis, and community-level causality. Established and emerging interventions-including probiotics, prebiotics, synbiotics, functional diets, biofloc management, phages, postbiotics, microbiota transplantation and synthetic microbial communities-are assessed according to their capacity to modify microbial function, persistence and recovery rather than taxonomic change alone. We further examine how multi-omics, microbiome-informed breeding, and environmental monitoring could support biomarker development, predictive decision support and context-specific intervention. We argue that progress requires a shift from taxonomic description to function-guided engineering, from endpoint comparisons to direct measurement of resilience, and from laboratory efficacy to reproducible farm-scale validation. Overall, microbiome management may contribute to more disease-resilient and sustainable shrimp production, provided that its effectiveness can be validated under commercial farming conditions.

Dysbiosis↗

Fertility in dogs in relation to semen quality and the time and site of insemination with fresh and frozen semen.

In this study 470 bitches were inseminated; 405 with fresh semen into the cranial vagina and 65 with frozen semen transcervically into the uterus. The pregnancy rate was 65.7% with fresh semen and 41.5% with frozen. When corrected for stage of oestrus at the time of insemination and for semen quality the pregnancy rate was 83.8% with fresh semen and 69.3% with frozen semen. The pregnancy rate improved with an increase in the number of inseminations. Inseminations with fresh semen before the time of ovulation (less than 17.3 nmol progesterone/l) were often successful. With frozen semen the peripheral plasma level of progesterone at the time of insemination was greater than 30 nmol/l in all but 1 of the bitches that became pregnant. This difference is assumed to be attributable to a longer survival time of several days for fresh semen than for frozen-thawed semen. Cytological scoring was a satisfactory method for determining the stage of oestrus when using fresh semen, but more precise methods are needed when using frozen semen. Semen quality was difficult to correlate with fertility. Pregnancies were obtained with fresh semen of inferior quality, although the litter size was smaller. With frozen semen no pregnancies resulted when the semen quality was poor. Litter size was estimated to be 21.5% smaller in bitches inseminated with fresh semen compared with naturally mated bitches. Litter size in bitches inseminated with frozen semen was 23.3% smaller than in bitches inseminated with fresh semen. Puppy deaths occurred in 35.5% of the litters in this study. The overall puppy death rate during the first 3 weeks of life was 11.9%. Although not statistically significant, there was a tendency for bitches treated with antibiotics at the time of mating or during early pregnancy to have a higher pregnancy rate and a larger litter size. There was also a tendency for breed differences in pregnancy rate, although not significant.

Animals↗

Anatomical study of cranial nerve emergence and skull foramina in the dog using magnetic resonance imaging and computed tomography.

Twenty-two magnetic resonance imaging (MRI) brain studies of different breeds of dogs were reviewed to assess the anatomy of cranial nerve (CN) origins and associated skull foramina. These included five anatomic studies of normal brains using 2-mm-thick slices and 17 studies using conventional clinical protocols with 3- or 4-mm slices on both normal and abnormal brains. Images were obtained in transverse, sagittal, and dorsal planes to allow a thorough comparison between studies. CNs II, III, V (and its divisions), and VIII were observed consistently on conventional studies. On the thin-slice studies, the origins and proximal portions of CNN IV, VII, and the group of IX, X, and XI could be seen. The origins of CNN VI and XII were not observed with certainty. In parallel, a computed tomography study of an isolated skull was performed with a thin copper wire within each of the skull foramina to determine precisely each CN exit and to facilitate recognition of the course of CNs when exiting the skull on MRI images.

Animals↗

Genetic and management factors that influence the susceptibility of cattle to Mycobacterium bovis infection.

Genetic variation in the susceptibility of cattle to Mycobacterium bovis infection exists in differences between families and species, but not breeds. Susceptibility to M. bovis infection increases with age of cattle. Natural exposure to M. bovis or environmental mycobacteria may assist in the development of specific immunity, but there is no direct evidence for such immunological priming of tuberculosis resistance in cattle. This has, however, been demonstrated in humans and other animals. Since non-specific mechanisms have a role in protective immunity, developing an effective vaccine will be difficult, even though some protection of other species has been achieved. Immunological suppression in the periparturient period can produce anergic reactors, which may act as a constant source of infection for cattle-to-cattle transmission. Circumstantial evidence suggests that an adequate intake of mineral, vitamin and protein reduces the susceptibility of cattle. Although weather patterns have been implicated in the susceptibility of herds to M. bovis infection, there is insufficient information to determine the risk factors precisely. It is concluded that some reduction in the susceptibility of cattle to M. bovis infection can be achieved by modifications to the management system to minimize risk factors, but that a considerable amount of further research is required.

Age Factors↗