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Biomedical subjects

W E Berndtson

Publications and source records attributed to W E Berndtson.

At least 19 recordsLinked to original sources

Plasma and testicular testosterone levels, volume density and number of Leydig cells and spermatogenic efficiency of rabbits.

Plasma and tissue testosterone concentrations were determined by radioimmunoassay in 12 eight-month-old sexually mature New Zealand White rabbits and evaluated for possible associations with spermatogenic efficiency as well as with volume density and number of Leydig cells. Testicular tissue was processed histologically and histometry was performed in order to quantify germ cells, Sertoli cells and Leydig cells. Spermatogenic efficiency, reported as the ratios among germ cells (spermatogonia, primary spermatocytes and round spermatids) and by the ratio of germ cells to Sertoli cells, was not associated with testosterone levels. However, Leydig cell parameters such as number of Leydig cells per gram of testis, total number of Leydig cells per testis and percent cell volume of Leydig cell nuclei were correlated significantly with testosterone levels. The statistically significant correlation (r = 0.82, P<0.05) observed between testosterone levels and the number of Leydig cells per gram of testis suggests that, in the rabbit, the latter parameter can serve as a criterion for monitoring testosterone levels in this species under normal conditions.

Animals↗

Assessing sex-drive in young Bos taurus bulls.

Objectives in this study were to determine the accuracy of different methods of evaluating libido score (L), service rate (SR) and reaction time to service (RTS) in yearling Bos taurus bulls. Using restrained, non-estrus females, 26 yearling Bos taurus bulls were evaluated a total of eight times (four sessions, two tests per session) over 2 consecutive months for L, SR, and RTS. Individual bull variation influenced both L (P<0.0001) and SR (P<0.003). Repeatability was moderate for L (R=0.64) and low for both SR (R=0.12) and RTS (R=0.04). Under the conditions of this study and characteristics of these tests, variance was reduced to 69-73% for L and 26-23% for SR with four to eight repetitions, respectively. Bulls that scored highly in the first session, however, tended to score highly throughout. Although, three bulls did not serve in any test, RTS was independent of effects. However, the tendency of RTS to decrease, and for L and SR to both increase with consecutive tests, suggests influences other than genetic, such as learning and/or environmental factors. These tendencies were most evident in bulls which obtained low L scores at the first session. It was concluded that, despite the important degree of individual variability observed in L and SR, accurate quantitative evaluation of the sex-drive in young Bos taurus beef bulls would benefit from use of test procedures of greater repeatability.

Analysis of Variance↗

Testicular weight, Sertoli cell number, daily sperm production, and sperm output of sexually mature rabbits after neonatal or prepubertal hemicastration.

The present study was conducted to investigate the influence of hemicastration and age at hemicastration on the subsequent testicular development of male rabbits through sexual maturity. Thirty New Zealand white rabbits were left intact or were hemicastrated on Day 35, 49, 77, or 105 postconception. Beginning at 7 mo of age, ejaculates were collected every other day for 1 mo, and the last ten ejaculates were used to quantify daily sperm output. At 8 mo, the rabbits were killed and their carcases, testes, testicular capsules, and epididymides were weighed. Testicular tissue was processed for quantification of sperm production rates by enumeration of homogenization-resistant spermatids and via histometric evaluation. Regardless of the age at hemicastration, this manipulation did not alter the development of the remaining testis as assessed from testis weight, numbers of Sertoli cells per testis, daily sperm production, or sperm output (p > 0.05). On the basis of these findings, it would appear that the development of the spermatogenic capacity of the remaining testis of the rabbit is not altered appreciably by hemicastration at a young age.

Animals↗

A simple, rapid and reliable method for selecting or assessing the number of replicates for animal experiments.

A simple approach was developed for determining the number of replicates needed per treatment group to provide experiments of known power and sensitivity, where power equals the probability that a treatment effect would not go undetected if an effect existed and sensitivity equals the minimal treatment response that will be detectable. This approach, in turn, was used to construct reference tables, applicable across scientific disciplines, from which researchers may read replication requirements directly with ease, speed and reliability. To use the tables, one need only furnish a reliable estimate of the coefficient of variability expected among replicates, which may be obtained from prior observations on similar populations. The tabular data also enable a rapid, reliable assessment of the actual power and sensitivity of completed experiments, such as those contained within the published literature.

Animals↗

Replication requirements and number of ejaculates needed for assessing treatment effects on sperm output and seminal characteristics of electroejaculated Holstein bulls.

Two ejaculates were harvested by electroejaculation on each of 3 d per week for 14 wk from 14, 12- to 24-mo-old Holstein bulls. Ejaculates during the first 2 wk served to stabilize sperm output. Data for the remaining ejaculates were used in a components of variance approach to determine the number of bulls per treatment and ejaculates per bull that would be needed to provide adequate sensitivity and precision for assessing treatment effects on seminal characteristics. The latter included the number of sperm (total and motile), volume and sperm concentrations of the sperm-rich fraction, and the percentage of progressively motile sperm in first and second ejaculates or per day. Replication requirements declined as the number of ejaculates per bull was increased to about 15 to 25 but declined minimally thereafter. The number of bulls needed per treatment varied in relation to the size of the treatment response to be detected but was much greater than the replication used in typical experiments. Replication for assessing sperm output in first ejaculates or per day was approximately one-half as great for experiments in which pretreatment information was used to adjust post-treatment data. Tables should be useful as a guide for designing efficient, cost-effective studies of known sensitivity and precision.

Animals↗

Optimal replication for histometric analyses of testicular function in rats or rabbits.

Quantitative evaluations of testicular histology can provide sensitive endpoints for determining toxicity of chemicals to the male reproductive system. But, the numbers of observations per testis or number of animals per treatment group often are selected by tradition or availability, rather than from a statistical basis. Therefore, we studied the number of observations per male (sampling intensity) and number of animals per treatment (replication) needed to detect treatment effects of given magnitude, with predictable error probabilities, using data from Sprague-Dawley rats and Dutch-belted rabbits that received 0.0, 0.94, 1.88, 3.75, 7.5, or 15.0 mg/kg body wt of 1,2-dibromo-3-chloropropane (DBCP). Data for one testis from 102 rats and 34 rabbits were available. For each testis, observations included measurement of the minor diameter of 50 seminiferous tubules, counts of the number of leptotene primary spermatocytes per 250 Sertoli cells, and counts of spherical spermatids within 20 seminiferous tubular cross sections. Tabular data are presented showing optimal numbers of observations per testis and animals per treatment group as a function of the difference to be detected and selected probabilities for Type I and II errors. In general, precise assessments required far fewer observations per testis than are used routinely. However, due to the inherent variability among animals, the number of animals required per treatment tended to be greater for experiments with the rabbit, and increased substantially for both species when detection of small differences had to be ensured. The data presented should enable investigators to design experiments of chosen sensitivity and precision while making cost-effective use of animals and labor.

Animals↗

Relationship of intratesticular testosterone content of stallions to age, spermatogenesis, Sertoli cell distribution and germ cell-Sertoli cell ratios.

Testes were obtained from 47 1-20-year-old stallions during the natural breeding season. Total testicular testosterone and testosterone/g testis increased with age (P less than 0.005), and total testicular testosterone was associated with larger testis size (P less than 0.05). Neither testosterone per gram nor per paired testes were related to total Sertoli cell number (P greater than 0.05), but greater testosterone per paired testes was associated with fewer Sertoli cells per unit of seminiferous tubule length (P less than 0.005) or basement membrane area (P less than 0.02) and with a higher number of germ cells supported per Sertoli cell (P less than 0.05). Although values for testosterone per gram and per paired testes were unrelated (P greater than 0.10) to sperm production/g testis or to the yield of spermatids/spermatogonium, testosterone per paired testes was positively related to sperm production per paired testes (P less than 0.05). It is concluded that intratesticular testosterone increases with age, is related in a positive manner to quantitative rates of sperm production, and can account for some of the differences in sperm production among individual stallions within a single breeding season.

Aging↗

Sampling intensities and replication requirements for detection of treatment effects on testicular function in bulls and stallions: a statistical assessment.

Data from testes of 16, 2- to 3-yr-old stallions and 34 yearling beef bulls were utilized in a components of variance approach to calculate the number of observations required per testis and(or) the number of animals required per treatment group to provide experiments of known sensitivity and precision, where treatment was to be assessed by one of several endpoints. The latter included paired testes weight, seminiferous tubular diameter, the number of germ cells per seminiferous tubular cross-section, or the number of elongated spermatids per gram of testicular parenchyma or per testis. For all variables for which several observations were available for each testis, precise assessment of any given male required far fewer observations than have been used routinely. However, replication requirements varied substantially in relation to the sensitivity (i.e., size of difference to be detected) and precision (i.e., Type I and II error probabilities) desired. Replication requirements were greater for stallions than for bulls, particularly at higher levels of sensitivity, for which requirements for both species were very large. The data presented should permit future experiments involving assessment of these endpoints to be designed with known sensitivity and precision and with optimal efficiency and cost-effectiveness.

Animals↗

Numbers of Sertoli cells, quantitative rates of sperm production, and the efficiency of spermatogenesis in relation to the daily sperm output and seminal quality of young beef bulls.

Data from 34 yearling Hereford or Angus bulls were used to investigate relationships of testicular size, quantitative rates of sperm production, Sertoli cell numbers, numbers of germ cells supported per Sertoli cell, and the efficiency of spermatogenesis to daily sperm output and seminal quality. Two ejaculates were collected by electroejaculation from each bull on each of 2 days/week throughout the study. The percentage of progressively motile sperm and the percentage of morphologically normal sperm were determined from aliquots of fresh semen. Additional aliquots of semen were frozen in glass ampules or plastic straws and subsequently evaluated for postthaw motility and percentage of sperm with intact acrosomes. Sertoli cell numbers, the numbers of germ-cells per Sertoli cell, and the efficiency of spermatogenesis were unrelated to the quality of fresh or frozen semen (P greater than 0.05). In first ejaculates, the numbers of sperm and motile sperm were related (P less than 0.05) to testicular parenchymal weight (r = 0.38 and 0.50), daily sperm production (r = 0.45 and 0.53), and spermatids per gram of testicular parenchyma (r = 0.35 and 0.34). Testicular parenchymal weight and daily sperm production also were related to daily sperm output and to the average daily motile sperm output of these bulls (P less than 0.05), but could account for less than 25% of the variability in these end points among bulls.

Animals↗

Spermatogenesis, sperm output and seminal quality of Holstein bulls electroejaculated after administration of oxytocin.

The 12- to 24-month-old Holstein bulls were electroejaculated twice on each of 3 days per week throughout the study. After a 2-week stabilization period and subsequent 2-week pre-treatment period, 7 bulls were given 50 i.u. oxytocin via the jugular vein 10 min before each first ejaculate for 10 weeks. The 7 control bulls were handled identically but did not receive oxytocin. All bulls were castrated at the end of the study. Oxytocin was without effect on spermatogenesis (P greater than 0.10). Oxytocin did not alter the total number of spermatozoa harvested per collection day (P greater than 0.10), but increased the number of spermatozoa in first ejaculates by an average of 34.2% (P less than 0.025). Oxytocin did not affect sperm quality (P greater than 0.10) as judged by the motility of spermatozoa in fresh semen or by the motility or percentage of spermatozoa with intact acrosomes in thawed semen. It is concluded that 50 i.u. oxytocin enhanced sperm output in first ejaculates of electroejaculated bulls without altering daily sperm production or seminal quality.

Animals↗

The numbers of Sertoli cells in mature Holstein bulls and their relationship to quantitative aspects of spermatogenesis.

Testes from 37 Holstein bulls, 38-99 mo of age, were used to investigate the relationship of Sertoli cell number, Sertoli cell-germ cell ratios and other related factors to daily sperm production (DSP). DSP was assessed by enumeration of spermatids in testicular homogenates, whereas Sertoli cell and germ cell ratios were based on direct counts in 20 round Stage VIII seminiferous tubular cross sections per bull. Numbers of Sertoli cells were calculated as (total homogenization resistant spermatids:spermatid:Sertoli cell ratio)/0.394; the factor of 0.394 adjusted for the presence of homogenization resistant spermatids during only 39.4% of the spermatogenic cycle. Data were subjected to simple linear and second-order regression analyses. Positive linear relationships were observed between DSP and testicular parenchymal weight (p less than 0.005, R = +0.71), DSP per gram (p less than 0.005, R = +0.79), total Sertoli cells (p less than 0.005, R = +0.83), Sertoli cells per gram (p less than 0.01, R = +0.47) and the yield of Step 8 spermatids per Type A spermatogonium (p less than 0.05, R = +0.34). DSP was not related (p greater than 0.10) to the number of germ cells supported per Sertoli cell. Testicular parenchymal weight and DSP per gram were unrelated to each other (p greater than 0.10), but both were related (p less than 0.005) to the total Sertoli cell number (R = +0.61 and +0.62, respectively). Total number of Sertoli cells accounted for more of the variation in DSP between bulls (R2 = 68.2%) than did any other factor examined. It was suggested that total Sertoli cell number may be an important determinant of a bull's spermatogenic potential.

Animals↗

Relationship of absolute numbers of Sertoli cells to testicular size and spermatogenesis in young beef bulls.

Testes were obtained from 34 Hereford or Angus bulls at about 1.5 yr of age and were used to investigate the relationship between the absolute number of Sertoli cells vs testicular size and daily spermatozoal production (DSP). Quantitative determination of DSP was based upon enumeration of elongated spermatids in testicular homogenates. The ratio of step 8 spermatids to Sertoli cells (S:SC) was established by direct counts of these cells in each of 20 round stage VIII seminiferous tubular cross sections for each bull. The number of Sertoli cells per paired testes was calculated as (total spermatids divided by S:SC)/.394, where total spermatids equalled the number of homogenization-resistant spermatids. The factor of .394 adjusted for the fact that the latter cells are present for only 39.4% of the spermatogenic cycle. All data were subjected to simple linear and second-order regression analyses. A positive linear relationship (P less than .005) was found between testicular weight (Y, in grams) and the absolute number of Sertoli cells per paired testes (X, in billions), which was characterized by the equation Y = 315.2 + 10.74X and a coefficient of correlation (r) of .56 (P less than .01). A similar relationship was observed between DSP (Y, in billions) and Sertoli cell numbers (X, in billions). This was characterized by the equation Y = 1.36 + .222X (P less than .005) and a coefficient of correlation of .70 (P less than .01). Daily sperm production was unrelated to the S:SC ratio (P greater than .05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Assessment of procedures for screening agents for effects on male reproduction: effects of dibromochloropropane (DBCP) on the rat.

This study was designed to evaluate new protocols proposed for use as an initial screen or a dose-response test of an agent for adverse effects on male reproduction. Seven groups of adult male rats (15/group) were nongavaged or received orally 0.00, 0.94, 1.88, 3.75, 7.5, or 15.0 mg/kg of 1,2-dibromo-3-chloropropane (DBCP) in corn oil each day for 77 days. From Day 65 to Day 71, each male was caged with 2 untreated female rats. Pregnancy rate and the ratio between the numbers of embryos and corpora lutea were determined. Males were killed on Day 78 and blood was collected and one testis and epididymis were used to determine daily spermatozoal production and epididymal spermatozoal reserves. The contralateral testis was fixed for quantitative histologic evaluation. Considering only data for the 15.0 and 0.00 mg/kg doses, as in an initial screen, DBCP would have been identified as a compound of concern. Body weight, paired testicular weight, parenchymal weight of the left testis, daily spermatozoal production per testis, numbers of sperm in the caput-corpus or cauda epididymidis, mean diameter of seminiferous tubules, and the ratio of leptotene spermatocytes to Sertoli cells were reduced (p less than 0.05). Fertility was normal, but the incidence of dead embryos and the ratio of dead embryos to corpora lutea were increased. Considering all data, as in a dose-response test, discriminant analysis was more sensitive in distinguishing treated rats from those receiving 0.0 mg DBCP/kg than was univariate analysis of variance. Using DBCP as a test compound, it was concluded that use of relatively simple, objective criteria in a test lasting approximately 10 weeks will be useful in identifying agents requiring detailed scrutiny.

Animals↗

Use of quantitative testicular histology to assess the effect of dibromochloropropane (DBCP) on reproduction in rabbits.

Dibromochloropropane (DBCP) was administered orally to 36 sexually mature male Dutch Belted rabbits assigned at random to one of six groups to receive 0, 0.94, 1.88, 3.75, 7.5, or 15.0 mg DBCP per kilogram of body weight daily 5 days per week during a 69-day treatment period. Animals were euthanized and necropsied on Day 70. Body weights and weights of the kidneys, liver, epididymides, and accessory sex glands were not influenced by DBCP treatment. The highest dosage reduced mean paired testes weight to 45% of control values (p less than 0.01). Mean seminiferous tubular diameter was reduced by 15 and 29% with the two highest dosages of DBCP, respectively (p less than 0.01), and the percentage of seminiferous tubules containing elongating and round spermatids, as the most advanced cell type present, was decreased (p less than 0.01). The number of leptotene primary spermatocytes per Sertoli cell, and the numbers of spermatogonia, young and old primary spermatocytes, and Step 1 spermatids per Stage I seminiferous tubular cross section likewise indicated a general depression (p less than 0.01) of spermatogenesis. Based upon the number of Step 1 spermatids per cross section at Day 70, production of sperm at this time by rabbits receiving the highest dose of DBCP was estimated to be less than one-fourth of control values. No significant effect (p greater than 0.05) of 0.94 mg DBCP/kg on any parameters was found, so this could be considered to be the no effect dose. However, the regressions and mean values are interpreted to indicate that the no effect level of DBCP may be less than 0.94 mg DBCP/kg of body weight. It is suggested that the most sensitive tests described here be included in future screening tests for potential effects of agents on male reproduction.

Animals↗

Dietary potentiation of the antifertility effects of 5-thio-D-glucose in male rats.

Male rats of proven fertility were fed the following diets for 28 d either with or without 0.075% 5-thioglucose (5-THG): AIN-76 diet (A76): a diet with 13% casein, 2% glucose and the balance of the calories as free corn oil fatty acids from corn oil (2G); and a similar diet, isocaloric with 2G, with the glucose level increased to 20% (20G). The diets alone without 5-THG had no influence on any of the parameters measured. Body weight gain was lower in rats fed diets containing 5-THG than in those fed diets without 5-THG. In rats fed A76, the only 5-THG effects on male reproductive tract (MRT) tissues was the appearance of testicular multinucleate giant cells (MGC). In rats fed either 2G or 20G, the MRT effects of 5-THG included the appearance of MGC, a lower number of germ cells at most stages of maturation, lower sperm counts and biochemical changes in testis slices and in germ cell preparations compared to rats not fed 5-THG. There were fewer Step 7 spermatids in rats fed 5-THG in 2 G than in those fed 5-THG in 20G. It is concluded that the MRT toxicity of 5-THG is influenced by diet, being potentiated by the low protein diet high in free fatty acids and, to a lesser extent, by low glucose levels within these diets.

Analysis of Variance↗

A quantitative study of Sertoli cell and germ cell populations as related to sexual development and aging in the stallion.

Testes from 47 stallions, 1-20 yr of age, were used to examine the influence of age on Sertoli and germ cell populations as well as on functional activity of Sertoli cells. For these stallions, the number of Sertoli cells per paired testes declined linearly with age, and was only 41.7% as great at age 20 as at age 2. However, development of reproductive organs proceeded until age 12-13, as evident from increases in paired testes weight and quantitative rates of spermatozoal production. Although the absolute number of Sertoli cells declined during this period of development, individual Sertoli cells displayed a remarkable capacity to accommodate greater numbers of developing germ cells. Between age 2 and age 12, the mean numbers of developing spermatogonia, young primary spermatocytes, old primary spermatocytes, and round spermatids supported by each Sertoli cell at Stage I of spermatogenesis increased by 49, 176, 153, and 161%, respectively.

Aging↗

Effect of dihydrostreptomycin or oxytetracycline on reproductive capacity of bulls.

Young beef bulls (n = 27) were used in a trial to study the effect of dihydrostreptomycin sulfate (DHS) or oxytetracycline (OTC) hydrochloride on spermatogenesis, epididymal sperm maturation, and freezability of sperm. Nine of the bulls were given a 22 mg/kg dose of DHS twice, 12 hours between doses. Nine other bulls were treated with OTC--1 dose of 26.4 mg/kg of body weight, and then 6 more doses each of 17.6 mg/kg, ca 12 hours between doses. The remaining 9 bulls were nontreated controls. The treatment regimens with the 2 antibiotics were without effect on spermatogenesis. These treatments also were without effect on seminal pH, ejaculate volume, percentage of motile spermatozoa, rate of spermatozoal motility, or concentration of spermatozoa in ejaculates harvested on day 3 or 7 of the study (day 0 = 1st day of treatment). There was a treatment-by-day effect on spermatozoal concentration; the number of sperm per milliliter was markedly increased on day 3 for OTC-treated bulls. The increased spermatozoal concentration in the OTC-treated group was associated with an influence of the antibiotic on ejaculation. All bulls given this antibiotic ejaculated without palpable penile engorgement or erection on day 3. On day 7 the rate of spermatozoal motility was increased in the 2 treatment groups compared with the rate in the control bulls. Also on day 7, the percentage of motile spermatozoa was greater in the OTC-treated bulls than in the control or DHS-treated bulls.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗