Effect on scrotal circumference of trucking beef bulls medium and long distances

Scrotal circumference (SC) was measured both before and after 2- to 3-yr-old beef bulls were trucked either medium (138 km) or long (565 km) distances. Two SC measurement techniques were used, one recommended by the Society for Theriogenology and the “slip-off” technique. No differences (P > 0.05) were observed between pre-and post- trucking SC measurements using the Theriogenology technique when bulls were trucked either medium (n = 12; 35.9 ± 0.8 vs 35.8 ± 0.7 cm) or long (n = 16; 36.5 ± 0.5 vs 36.6 ± 0.4 cm) distances. However, when the SC of the same bulls was measured using the “slip-off” technique, there was a small, but significant (P < 0.05), reduction in SC for bulls trucked medium (38.0 ± 0.7 vs 37.5 ± 0.7 cm) and long (38.9 ± 0.4 vs 38.6 ± 0.4 cm) distances. Bulls having SC at or below the recommended minimum standard reacted no differently to the stress of trucking than did bulls having a SC well above the standard. Errors in measurement technique, severe weather conditions and testicular degeneration over an extended interval are discussed in terms of their potential contribution to the reduction in SC observed by some cattlemen. These data support the use of the SC measurement technique recommended by the Society for Theriogenology because its accuracy is not affected by trucking of bulls. The suggested implementation of an adjustment period between the trucking of beef bulls and measurement of their SC using the Theriogenology technique is not supported.     

Echocardiography practice, training and accreditation in the intensive care: document for the World Interactive Network Focused on Critical Ultrasound (WINFOCUS)

Echocardiography is increasingly used in the management of the critically ill patient as a non-invasive diagnostic and monitoring tool. Whilst in few countries specialized national training schemes for intensive care unit (ICU) echocardiography have been developed, specific guidelines for ICU physicians wishing to incorporate echocardiography into their clinical practice are lacking. Further, existing echocardiography accreditation does not reflect the requirements of the ICU practitioner. The WINFOCUS (World Interactive Network Focused On Critical UltraSound) ECHO-ICU Group drew up a document aimed at providing guidance to individual physicians, trainers and the relevant societies of the requirements for the development of skills in echocardiography in the ICU setting. The document is based on recommendations published by the Royal College of Radiologists, British Society of Echocardiography, European Association of Echocardiography and American Society of Echocardiography, together with international input from established practitioners of ICU echocardiography. The recommendations contained in this document are concerned with theoretical basis of ultrasonography, the practical aspects of building an ICU-based echocardiography service as well as the key components of standard adult TTE and TEE studies to be performed on the ICU. Specific issues regarding echocardiography in different ICU clinical scenarios are then described. Obtaining competence in ICU echocardiography may be achieved in different ways – either through completion of an appropriate fellowship/training scheme, or, where not available, via a staged approach designed to train the practitioner to a level at which they can achieve accreditation. Here, peri-resuscitation focused echocardiography represents the entry level – obtainable through established courses followed by mentored practice. Next, a competence-based modular training programme is proposed: theoretical elements delivered through blended-learning and practical elements acquired in parallel through proctored practice. These all linked with existing national/international echocardiography courses. When completed, it is anticipated that the practitioner will have performed the prerequisite number of studies, and achieved the competency to undertake accreditation (leading to Level 2 competence) via a recognized National or European examination and provide the appropriate required evidence of competency (logbook). Thus, even where appropriate fellowships are not available, with support from the relevant echocardiography bodies, training and subsequently accreditation in ICU echocardiography becomes achievable within the existing framework of current critical care and cardiological practice, and is adaptable to each countrie's needs.     

Differences in bull spermiograms using eosin-nigrosin stain, feulgen stain, and phase contrast microscopy methods

The Society for Theriogenology recently adopted a minimum standard of 70% normal spermatozoa for the bull breeding soundness examination (BSE). We conducted this study to determine if spermiograms derived by brightfield microscopy of eosin-nigrosin stained semen smears (Society method) overestimated the proportion of normal spermatozoa. Comparison of the above method was made with that of phase contrast microscopy (Phase method). We then evaluated our ability to discern head abnormalities by comparing brightfield microscopy of Feulgen-stained sperm DNA (Feulgen method) with those of the Society and Phase methods. Spermiograms were determined for each of the 181 beef bulls using all 3 methods. Only bulls that were being routinely tested prior to the 1993 breeding season were included. The mean percentage of normal spermatozoa surpassed the minimum standard with the Society (72.8%) but not the Phase (52.5%) method, which identified more distal cytoplasmic droplets that adhered to cells without distal midpiece reflexes. The Phase method also identified more total primary and fewer primary head abnormalities. We conclude that the Phase method is not a suitable substitute for the Society method when applying the minimum standard during a routine BSE. The Feulgen method identified more head abnormalities, especially in the pattern of DNA, than the other methods, however, when compared to the minimum standard that improvement was not clinically important. Both the Phase and Feulgen methods are better than the Society method for monitoring changes in abnormal spermiograms over time.     

Breeding soundness evaluations of 3,648 yearling beef bulls using the 1993 Society for Theriogenology guidelines

Our objective was to perform a retrospective analysis of breeding soundness evaluations (BSEs) as classified by the 1993 Society for Theriogenology (SFT) guidelines [Chenoweth et al., Guidelines for using the bull breeding soundness evaluation form, in: Theriogenology Handbook, 1993, pp. B-10]. Data included BSE information obtained from five performance-testing stations in South Carolina (SC1, SC2, SC3) and Tennessee (TN1, TN2) from 1986 through 1999 on 3648 Angus, Brangus, Charolais, Chianina, Gelbvieh, Limousin, Polled Hereford, Santa Gertrudis, Simbrah, and Simmental bulls. Analyses were simplified by classifying all bulls as either satisfactory or unsatisfactory potential breeders. Of the 3648 bulls evaluated, 76.2% were classified as satisfactory potential breeders. Of all bulls evaluated, 4.0% were unsatisfactory due to inadequate spermatozoal motility, 7.0% due to inadequate spermatozoal morphology and 2.6% due to a combination of inadequate motility and morphology. Unsatisfactory classifications due to non-spermatozoal parameters out of all bulls were 10.2%, with 7.1% for inadequate scrotal circumference and 3.1% for physical abnormalities. For satisfactory and unsatisfactory bulls, respectively, means and standard deviations were 35.8 +/- 2.7 and 33.0 +/- 4.1 cm (P < 0.001) for scrotal circumference, 63 +/- 18 and 35 +/- 24% (P < 0.001) for percent motility, and 86 +/- 7 and 63 +/- 21% (P < 0.001) for percent normal morphology.     

The development of beef breeding bulls

Management of the bull battery will have a dramatic impact on profitability of the cow/calf enterprise. It is critical that young bulls be selected and developed to maximize longevity and productivity for the eventual buyer. Bulls must be structurally sound, healthy, and have adequate libido in order to service the required number of females. Once bulls complete their first breeding season, special care must be taken in order to ensure that they recover and regain needed body condition and pass a bull breeding soundness examination (BBSE). Mature bulls that have reached their genetic potential for growth require less intensive management, but the health program and annual BBSE cannot be overlooked. Mature bulls are also more likely to carry venereal disease and should be screened according to local disease incidence and state regulations. All bulls, regardless of age, should be observed early during the breeding season to ensure that they are physically capable of mounting and servicing females. The establishment of a complete management program, especially for young bulls, is essential to ensure that ranch resources are used efficiently, including maintenance of a high level of reproductive performance of the cow herd.     

The cow as an induced ovulator: Timed AI after synchronization of ovulation

Timed-AI after synchronization of ovulation has become one of the most used reproductive technologies developed during the past 40 years. Various adaptations of this technology are now extensively used worldwide, in the beef and dairy cattle industry. Our well-cited report, published in Theriogenology in 1995, presented a method termed Ovsynch, that used GnRH and PGF_2α to perform synchronization of ovulation and timed AI in lactating dairy cows. This report introduced Ovsynch, more as a concept of induced ovulation, and demonstrated the ovarian dynamics during the protocol. Validation and improvements on this method were subsequently performed in numerous university studies and on commercial dairies, worldwide. This review will provide a brief historical background, some personal recollections, and certain modifications that have been made in synchronization of ovulation protocols. Each section emphasizes the physiology that underlies the most widely-used synchronization of ovulation protocols and key modifications and some practical application of these protocols on commercial operations. Finally, the effect of timed AI in the US dairy industry and in the Brazilian beef cattle industry are compared. Although numerous studies have been done using these protocols, there is still substantial need for research to improve the synchronization, efficacy, simplicity, and practical application of these protocols.     

Scrotal circumference measurements in 764 beef bulls.

A total of 764 breeding soundness examinations was conducted on beef bulls utilizing the method of examination and criteria for classifying bulls of the Society for Theriogenology. In addition to this examination each bull was subjected to scrotal circumference measurement and to weighing. Classification of the bulls according to breeding soundness potentials was as follows: 88% of the bulls were satisfactory potential breeders, 8% of the bulls were questionable potential breeders and 4% of the bulls were unsatisfactory potential breeders. The proportions of bulls in each classification; satisfactory, questionable, or unsatisfactory; were not different among the four breeds evaluated. The majority of bulls evaluated in this study were between 14 and 36 months of age and weighed between 900 and 1500 pounds. For the ages and weights evaluated, scrotal circumference measurement variances were not closely related to age and weight differences. There was a tendency shown for “Questionable” and “Unsatisfactory Potential Breeders” to have smaller scrotal circumference measurements. The study indicates that Angus, Charolais, Horned Hereford and Polled Hereford bulls of breeding ages and weights should have scrotal circumference measurements of at least 32 centimeters in order to be classified as “Satisfactory Potential Breeders”.     

Outcomes of breeding soundness evaluation of 2898 yearling bulls subjected to different classification systems

This study is a retrospective analysis comparing data on yearling bull breeding soundness evaluation (BSE) subjected to 3 different classification systems: the Society for Theriogenology (SFT) 1983 and 1993 systems, and the Western Canadian Association of Bovine Practitioners (WC) 1993 system. Data were collected at 5 performance bull-test stations located in South Carolina and Tennessee from 1986 through 1996. Yearling bulls (n=2898) that were 10 to 20 mo of age were used in the analysis. To simplify analysis, bulls were determined to be either satisfactory or unsatisfactory potential breeders (including those classified as questionable, deferred or unsatisfactory). Data were analyzed 1) within location where a location was treated as an individual experiment, 2) with breeds and locations collapsed, and 3) within age-group where each age-group was treated as an individual experiment. An ANOVA was performed using GLM procedures of SAS, and this model was used to generate least square means for the proportion of bulls classified as satisfactory and the 5 possible unsatisfactory outcomes due to inadequacies in scrotal circumference, spermatozoal morphology, spermatozoal motility, a combination of inadequate spermatozoal morphology and motility, or physical abnormalities. Inadequate scrotal circumference or physical abnormality did not affect differences for BSE outcomes among systems. Using the SFT93 system, bulls failed at a higher rate due to inadequate spermatozoal morphology (P < 0.05) than when subjected to the SFT83 system. In the WC93 system, a higher percentage of bulls failed due to inadequate spermatozoal motility and to a combination of inadequate spermatozoal morphology and motility than in the other 2 systems (P < 0.05). The proportion of bulls in this data failing under the WC93 system appears to be the result of overestimation.     

Solid phase synthesis of peptide hydroxamic acids on poly(ethylene glycol)‐based support

A novel resin designed for solid‐phase synthesis of peptide hydroxamic acids (PHA) combining the trityl linker with poly(ethylene glycol)‐based support, ChemMatrix® type, is described. The synthesis of PHA can be performed according to a standard protocol, providing products in excellent purity and reasonable yields. Copyright © 2012 European Peptide Society and John Wiley & Sons, Ltd.     

PIG3V, an immortalized human vitiligo melanocyte cell line, expresses dilated endoplasmic reticulum

Summary Vitiligo is an enigmatic pigmentary disorder of the skin. Factors potentially involved in the progressive loss of melanocytes from the basal layer of the epidermis include genetically determined aberrancies of the vitiligo melanocyte. It follows that analysis of melanocytes cultured from vitiligo donors can contribute to a further understanding of the etiopathomechanism. A setback for vitiligo research has been the limited availability of vitiligo-derived melanocytes. To overcome this limitation, we have generated a vitiligo melanocyte cell line according to a protocol established previously for the immortalization of normal human melanocytes. Vitiligo melanocytes Ma9308P4 were transfected with HPV16 E6 and E7 genes using the retroviral construct LXSN16E6E7. Successful transformants were selected using geneticin and subsequently cloned to ensure genetic homogeneity. The resulting cell line PIG3V has undergone more than 100 cell population doublings ince its establishment as a confluent primary culture, whereas untransfected melanocytes derived from adult skin senesce after a maximum of 50 population doublings. Cells immortalized by this transfection procedure retain lineage-specific characteristics and proliferate significantly faster than parental cells. In this study, the phenotype of PIG3V resembled melanocytes rather than melanoma cells in culture. Tyrosinase was processed properly and melanosomes remained pigmented. Importantly, ultrastructural characterization of PIG3V cells revealed dilated endoplasmic reticulum profiles characteristic of vitiligo melanocytes. An explanation for this dilation may be found in the retention of proteins with molecular weight of 37.5, 47.5, and 56.5 kDa, as determined by gel electrophoresis of microsomal proteins isolated from radiolabeled cells. Presented in part at the Annual Meeting of the Panamerican Society for Pigment Cell Research, Aspen, Colorado, 1998.     

Psychiatry in General Practice

Psychiatrists should include the family doctor in their plans for future psychiatric services. The general practitioner now treats most of the patients who seek help for psychiatric disorder and he could not give up his psychiatric practice even if he wanted to. Furthermore, there are not now nor will there ever be enough psychiatrists to take over all patients with mental ills. Most emotionally disturbed patients can be better handled by their family physicians than by a specialist.To provide the best care for emotionally disturbed people the communication between family doctors and psychiatrists must be improved. The specialist must acknowledge the importance of the general practitioner''s role in psychiatric diagnosis and treatment and give him more help. Medical schools must provide better undergraduate and postgraduate psychiatric training for the students who will become family doctors. Health plans and other prepayment agencies should properly compensate the general practitioner for giving psychiatric treatment. The specialist in psychiatry should consult more readily with the general practitioner and help him carry out some of the therapy. General hospitals should permit family doctors to admit mental patients to psychiatric wards in a general hospital and to carry out psychiatric treatment with the help of the specialist in psychiatry.     

Cell biology and functional dynamics of the mammalian sperm surface

Theriogenology has now a 40-year rich history on covering sperm biological aspects with a special emphasis on farm and husbandry animals. The major and most influential of these contributions will be placed into an evolutionary perspective of ongoing and intriguing progresses made in this field. Although many molecular details have been published, it is more the aim of this contribution to provide a guide through the main established aspects and concepts of sperm surface biology and refer only to major molecular players and mechanisms involved in sperm physiology. Those interested in more molecular details and in-depth knowledge can easily access the most relevant literature which is included here for reference purposes. With this approach, a logical and easy to follow buildup can be made of the general picture of sperm surface dynamics and of the ergonomics of sperm physiology and their function in mammalian fertilization. Understanding the ins and outs of sperm surface biology and the dynamics thereof, might challenge future researchers to design novel generation of better sperm-handling procedures. This could be beneficial for assisted reproductive technology and animal breeding industries.     

Non-reducing alkaline solubilization and rapid on-column refolding of recombinant prion protein

Mature prion protein (PrP) is a 208-residue polypeptide that contains a single disulfide bond. We report an alternative method to purify recombinant mouse PrP produced in Escherichia coli. Bacterial inclusion bodies were solubilized in a buffer containing 2 M urea at pH 12.5. The solubilized protein was rapidly purified on a nickel affinity column without a chaotrope gradient, followed by ion-exchange chromatography. The yield and purity of PrP produced by this alternative approach was similar to that obtained using a conventional solubilization and on-column refolding protocol. Recombinant PrP produced using the non-reducing purification protocol is properly folded, as determined by circular dichroism, and a competent substrate for amyloid fibril formation, as determined by Thoflavin-T dye binding assays. In summary, this report describes a rapid method for producing properly folded recombinant PrP without reducing agents or a chaotrope gradient.     

Who is publishing in Theriogenology and on what subjects?

An analysis of 503 articles published in the journal Theriogenology in 1984, 1987 and 1990 was conducted to identify research trends and researchers publishing in the journal. A 30.5% increase in the number of papers published was found for 1990. During the 3yr period studied, the United States and Canada accounted for 63.1% of the total number of articles published, followed by European countries with 14.3%, and by Asia with 12.0%. At least 1 paper was published from 41 countries. The United States, at the top of the list, increased its share from 16% in 1984 to 21% in 1990. The number of papers from Japan increased significantly while the percentage of participation from Europe declined. The female animal was the sole subject of 63.2% of the articles, and cattle were the most researched species, appearing in 44.3% of the studies. Although the estrous cycle was the most studied reproductive event in all 3 yr, there was also a trend for the popularity of this topic to decrease in favor of the subject fertilization. The number of authors per paper increased significantly over the years analyzed, as did the number of studies carried out in international cooperation. Institutes of higher education were represented in almost 70% of the papers, followed by research institutes (12.8%). The present analysis confirms the central role that the journal Theriogenology plays in the world-wide communication of research results in animal reproduction.     

Effects of the anabolic steroid, boldenone undecylenate, on certain reproductive parameters in bulls

A total of 17 bulls was used to study the effects of boldenone undecylenate on growth and semen characteristics in beef bulls. In trial 1 nine mature mixed-breed beef bulls with satisfactory semen quality were divided into two groups. Group I (n = 5) received boldenone undecylenate (1.1 mg/kg) at 21 d intervals for a total of seven treatments (147 d). Group II (n = 4) served as untreated controls. Semen was collected from each group by electroejaculation on each treatment day and evaluated according to the standards of the Society for Theriogenology. Although neither the percentage of spermatozoa with primary or secondary morphological abnormalities was different, the ejaculates of Group I bulls contained a higher percentage of abnormal spermatozoa than those in Group II. In trial 2, eight mixed-breed bull calves, average weight 140.4 kg, were maintained under drylot conditions in a single paddock. The bulls were divided into two equal groups. Group I (n = 4) received boldenone undecylenate as in Trial 1. Group II (n = 4) served as untreated controls. The bulls were weighed and the scrotal circumference (SC) was measured every 21 d until it reached 30 cm, at which time semen was collected and evaluated as in Trial 1. Group I bulls had a higher percentage of spermatozoa with primary morphological abnormalities than bulls in Group II. Group I bulls had a higher average daily gain (ADG) than Group II bulls and required 21 d longer for the SC to reach 30 cm. Semen quality for all bulls was satisfactory at each sampling day.     

Isolation of a slowly adhering cell fraction containing stem cells from murine skeletal muscle by the preplate technique

This protocol details a procedure, known as the modified preplate technique, which is currently used in our laboratory to isolate muscle cells on the basis of selective adhesion to collagen-coated tissue culture plates. By employing this technique to murine skeletal muscle, we have been able to isolate a rapidly adhering cell (RAC) fraction within the earlier stages of the process, whereas a slowly adhering cell (SAC) fraction containing muscle-derived stem cells is obtained from the later stages of the process. This protocol outlines the methods and materials needed to isolate RAC and SAC populations from murine skeletal muscle. The procedure involves mechanical and enzymatic digestion of skeletal muscle tissue with collagenase XI, dispase and trypsin followed by plating the resultant muscle slurry on collagen type I-coated flasks where the cells adhere at different rates. The entire preplate technique requires 5 d to obtain the final preplate SAC population. Two to three additional days are usually required before this population is properly established. We also detail additional methodologies designed to further enrich the resultant cell population by continuing the modified preplating process on the SAC population. This process is known as replating and requires further time.     

An Optimized Protocol of Protargol Staining for Ciliated Protozoa

Protargol staining is a crucial method to reveal the infraciliature of ciliates, which is the most important morphological character for species identification. In the present study, Wilbert's protocol of protargol staining was emended mainly toward the highly happened improper bleaching. Through reciprocal treatments, both insufficient and excessive bleachings were much eliminated from the protargol protocol and the tests performed with four different species of ciliates established that the stainings were considerably improved and more reliable with optimized bleaching. Compared to the original protocol, the optimized method was proved to be more suitable for the groups difficult to stain, and it is also friendlier for the beginners and researchers in related fields.     

Reliable detection of paternal SNPs within deletion breakpoints for non-invasive prenatal exclusion of homozygous α-thalassemia in maternal plasma

Reliable detection of large deletions from cell-free fetal DNA (cffDNA) in maternal plasma is challenging, especially when both parents have the same deletion owing to a lack of specific markers for fetal genotyping. In order to evaluate the efficacy of a non-invasive prenatal diagnosis (NIPD) test to exclude α-thalassemia major that uses SNPs linked to the normal paternal α-globin allele, we established a novel protocol to reliably detect paternal SNPs within the (--(SEA)) breakpoints and performed evaluation of the diagnostic potential of the protocol in a total of 67 pregnancies, in whom plasma samples were collected prior to invasive obstetrics procedures in southern China. A group of nine SNPs identified within the deletion breakpoints were scanned to select the informative SNPs in each of the 67 couples DNA by multiplex PCR based mini-sequencing technique. The paternally inherited SNP allele from cffDNA was detected by allele specific real-time PCR. A protocol for reliable detection of paternal SNPs within the (--(SEA)) breakpoints was established and evaluation of the diagnostic potential of the protocol was performed in a total of 67 pregnancies. In 97% of the couples one or more different SNPs within the deletion breakpoint occurred between paternal and maternal alleles. Homozygosity for the (--(SEA)) deletion was accurately excluded in 33 out of 67 (49.3%, 95% CI, 25.4-78.6%) pregnancies through the implementation of the protocol. Protocol was completely concordant with the traditional reference methods, except for two cases that exhibited uncertain results due to sample hemolysis. This method could be used as a routine NIPD test to exclude gross fetal deletions in α-thalassemia major, and could further be employed to test for other diseases due to gene deletion.     

Canine pyometra

Pyometra, literally meaning pus in the uterus, is a common disease entity of intact bitches. Similar conditions occur, including hydrometra and mucometra [De Bosschere H, Ducatelle R, Vermeirsch H, Van Den Broeck W, Coryn M. Cystic endometrial hyperplasia-pyometra complex in the bitch: should the two entities be disconnected? Theriogenology 2001;55:1509-19]. The exact etiology is unknown; however the repeated and prolonged response to estrogen followed by long intervals of progesterone dominance in the intact bitch leads to hormonally mediated changes in the endometrium. The endometrium changes when impacted by bacterial infiltration; changes in endometrial steroid receptors can result in the clinical syndrome described as pyometra. This paper will describe the signalment, risk factors, prevalence, proposed etiologic events, and both medical and surgical therapies. In addition, the prognosis for successful outcome and effects on future reproduction will also be described.     

Assessment of the uncertainty of trace metal and nitrogen concentrations in mosses due to sampling,sample preparation and chemical analysis based on the French contribution to ICP-Vegetation

Moss bio-monitoring is a convenient tool for establishing specific or large-scale exposure to atmospheric trace metals and nitrogen pollution. However, the uncertainty associated with sampling, sample preparation and chemical analysis of bio-monitors has been poorly documented, with the exception of one study dealing with lichens; thus, the uncertainty associated with moss bio-monitoring has never been assessed. Here, we propose following the Eurachem guidelines to determine the uncertainties associated with the concentrations of elements measured in mosses during the sampling survey across France in 2011. In addition, we assess the analytical method used in four surveys from 1996 to 2011. Uncertainties were expressed as linear functions of the element concentrations, with minimum and maximum slopes of 14 and 61%, corresponding to nitrogen and chromium, and a median of 32%. Although the data reveal that some steps of the protocol should be performed carefully, they also indicate that the protocol was executed properly and is reproducible. The chemical analyses contribute to a small proportion of the uncertainty associated with the protocol, except for the analysis of chromium. The sampling period, and intra-site variability largely contribute to this uncertainty. The moss species did not introduce additional uncertainty. This integrative assessment of measurement uncertainty will help improve the protocol for future surveys. Such studies could be useful for developing a standard operating procedure and could be used to improve comparisons between countries and to identify temporal trends across Europe.