McCartney, H. K.

Stallcup, O. T.

English

UARK (University of Arkansas )

Journal of the Arkansas Academy of Science Volume 6 Article 11 1953 Motility of Spermatazoa and Control of Bacteria in Bovine Semen Extenders Containing Sulfanilamide, Aureomycin, Terramycin, Bacitracin, Penicillin, and Strepromycin H. K. McCartney University of Arkansas, Fayetteville O. T. Stallcup University of Arkansas, Fayetteville Follow this and additional works at: https://scholarworks.uark.edu/jaas  Part of the Other Animal Sciences Commons Recommended Citation McCartney, H. K. and Stallcup, O. T. (1953) "Motility of Spermatazoa and Control of Bacteria in Bovine Semen Extenders Containing Sulfanilamide, Aureomycin, Terramycin, Bacitracin, Penicillin, and Strepromycin," Journal of the Arkansas Academy of Science: Vol. 6 , Article 11. Available at: https://scholarworks.uark.edu/jaas/vol6/iss1/11 This article is available for use under the Creative Commons license: Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0). Users are able to read, download, copy, print, distribute, search, link to the full texts of these articles, or use them for any other lawful purpose, without asking prior permission from the publisher or the author. This Article is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Journal of the Arkansas Academy of Science by an authorized editor of ScholarWorks@UARK. For more information, please contact scholar@uark.edu. MOTILITY OF SPERMATOZOA AND CONTROL OF BACTERIAIN BOVINE SEMEN EXTENDERS CONTAINING SULFANILAMIDE,AUREOMYCIN, TERRAMYCIN. BACITRACIN, PENICILLIN, AND STREPTOMYCINH. K. McCARTNEY and 0. T. STALLCUP 1University of ArkansasThe importance of controlling bacterial populations in bovine semen usedfor artificial insemination has been pointed out by Salisbury et al. (15). Asignificant relationship has been found between the number of bacteria in di-luted semen and its fertility when all types of bacteria were considered (4).The growth of many organisms commonly found in bovine semen is reportedly in-hibited by the addition of sulfonamides (6, 9, 10, 13, 16), of penicillin (1, 8,9, 18), of streptomycin (2, 18), of streptomycin plus penicillin (3, 12), ofpolymyxins (6), and of aureomycin (6, 11, 18), without significant toxic effectson. the motility of spermatozoa. However, no data have been reported on the con-trol of bacterial growth in semen by the use of bacitracin or terramycin.Finlay et al. (5) have reported that terramycin is bacteriostatic to manyGram-negative enteric organisms, aerobic spore formers, and Gram-positive cocci.It also has antirickettsial activity and appears to inhibit certain influenzaviruses.Rose (14) has reported that bacitracin is effective against most strains ofhemolytic streptococci, nonhemolytic streptococci, coagulase positive staphly-ococci, pneumococci , gonococci, anaerobic cocci in general, certain bacilli,certain spirochetes, actinomycetes and some protozoa. It shows little actionagainst the large group of aerobic Gram-negative nonsporeforming bacilli. Inview of the reported high bactericidal activity of terramycin and bacitracin, itseemed desirable to study the control of bacterial growth and effects on mo-tility of the spermatozoa by these compounds as compared to previously usedbactericidal agents.Materials and Methods. The antibacterial agents used in these investiga-tions are shown in Table 1.TAELE 1. Levels of Antibacterial Agents Used per 100 ml of Citrate-yolk ExtenderLPeni-Aureo-< i11in mycin Dihydro- TerramycinSulfa- G hydrochol- Bacitracin strepto- hydrochol-nilamide sodium ride (55 units mycin rideExtender (mgm) (mgn) (mgm) per mgn) (mgm) (mgn)Control ----0 0 0 0 0 0A 300 30 10 10 100 10BC-- --10D- -. 300 30 - -- 100E--10Extender D was used for comparison because of the common usage of theseantibacterial agents in extenders. Extender A was used to determine ifa combi-nation of all the antibiotics studies was desirable. To make a simultaneouscomparison of the effects of the different antibiotics on the motility of sperm-The authors are indebted to Herbert G. Luther, Charles Pfizer and Company, Incorporation,for the terramycin hydrochloride used in these investigations, and to Gertrude M. Cox, direc-tor, Institute of Statistics, University of North Carolina, for assistance in outlining some ofthe statistical procedures used in the analysis of the data.Research Paper 1070 Journal Series. University of Arkansas*39Journal of the Arkansas Academy of Science, Vol. 6 [1953], Art. 11Published by Arkansas Academy of Science, 195340 ARKANSAS ACADEMY OF SCIENCEatozoa and upon bacterial control, semen samples obtained from the ArkansasArtificialBreeders' Association of Fayetteville, Arkansas, were divided intoas many equal portions as there were antibacterial agents to be tested. Eachportion of semen was mixed with 3 per cent citrate-yolk extender to which theantibiotics, or antibiotics plus sul fani 1amide, had been added. The averagedilution rate was one part of semen to 40 parts of citrate-yolk extender. Theextended semen then was divided into equal portions and placed in 9-ml tgsttubes filled to capacity. One portion was stored in a refrigerator at 5 Cwhile the other portion was exposed to a temperature of 30° C ± 2° C. Dailymicroscopic examinations of the diluted semen afforded an estimate of the ef-fects of the antibacterial agents on the motility of the spermatozoa. Averagemotility readings, after 48 hours of storage, are presented as this time in-terval and probably represent a near maximum storage time during which semen isused by most artificial breeding associations in this country. The samples wereread until the motility reading fell below 30 per cent and then discarded.Statistical significance between means was tested by analysis of variance (17).The number of living bacteria was determined by the plate count method.After incubation at 37° C for 48 hours, the number of colonies on each platewas counted and the number of bacteria that would have been present in an en-tire milliliterof the diluted semen was calculated. Control plates were madeto check the sterility of the water used for dilutions, of the agar, and of theatmosphere in the laboratory during the plating procedure.Results. A summary of the data concerning the spermatozoa motility, theduration of motility, and the number of bacteria in extended semen containingthe antibacterial agents studied is presented in Table 2. After 48 hours ofstorage at 5* C, bacterial growth was not excessive in any of the extended semensamples. The combination of all antibacterial agents usually inhibited bacteri-al growth completely. Bacitracin and aureomycin completely inhibited bacterialgrowth in 50 per cent of the samples, this difference in bactericidal actionapparently resulting from the predominating types of bacteria in differentsamples.TABLE 2. Motility of Spermatozoa, Duration of Motility, and Numbers of Bacteriain Extended Semen Containing Various Antibacterial AgentsDurationStorage After 48 hours storage motilityTemper- motile bacteria rating ofatures spermatozoa 1 per ml. 2 30 per cent 2°C per cent Thousands HoursControl 5° 39.8 5.0 145.330* 9.6 415.7 28.0A (Combination of anti- 5° 48.7 .3 181.3bacterial agents) 30° 32.8 2.5 85.3B (Terramycin hydrochlo- 5° 46.0 6.5 154.7ride) 30° 19.9 5.2 42.7C (Bacitracin) 5° 48.1 .8 180.030° 7.9 474.7 38.0D (Penicillin / strepto- 5° 44.1 5.1 173.3mycin / sulfanilamide) 30° 33.5 15.2 77.3E (Aureomycin hydrochlo- 5° 40.6 .7 156.0ride) 30* 28.0 18.2 58.7'Average of 10 ejaculates.Average of nine ejaculatesAt 30° C bacteria multiplied rapidly when no antibacterial agent was pres-ent as was the case when bacitracin was present, this antibacterial agent ap-parently being inactivated at the higher temperature. Putrefaction of the eggyolk consistently occurred when bacterial growth was high. The percentage of40Journal of the Arkansas Academy of Science, Vol. 6 [1953], Art. 11https://scholarworks.uark.edu/jaas/vol6/iss1/11MOTILITY OF BOVINE SPERMATOZOA 41samples exhibiting a putrid odor after 48 hour's storage at 30° C was 40, and30, respectively, for the extenders containing bacitracin and no antibacterialagents, respectively.The analyses of variance of the percentage of progressively motile sperma-tozoa after 48 hour's storage is presented in Table 3. At 5° C the percentageof motile spermatozoa was similar in all extenders throughout the 48 hour'sstorage period. The effects of temperature appear to be significant, all sam-ples showing a lower average motility after 48 hours at 30* C as compared to 5°C, regardless of the diluter used (P<0.01). Similar results for a 72 hourstorage interval were reported by Foote and Bratton (7). At 30° C after a 48hour storage period significant differences between extenders were observed(P<0.01). Aureomycin, penicillin / streptomycin / sul fanilamide, and a combi-nation of antibacterial agencs were significantly superior (P<0.01) to noantibacterial agent and bacitracin at this temperature. Bacitracin, terramycin,and no antibacterial agent were not significantly different from each other(P<0.05). This was probably caused by a high bacterial count in samples con-taining bacitracin, and the low motility of the samples containing terramycinmay have resulted from toxicity although no such effects were evident when sam-ples were stored at 5° C.An analysis of variance of the data concerning the length of time a pro-gressive motility rating of 30 per cent was maintained is presented in Table 4.The temperature effects were again significant, the duration of motility beingappreciably lower in samples stored at 30* C as compared to those stored at 5°C. The fact that the extender effect at 30* C is riot significant suggests thatthe criterion of response used here is not as precise as the one used in Table3. This is probably because the spermatozoa died rapidly in some extendersstored at 30° C, and daily microscopic examination would not detect preciselywhen 30 per cent of the spermatozoa were motile.TABLE 3. Analysis of Variance of Percentages of Progressively MotileSpermatozoa after 48 hour's StorageFactor S.S. D.F. M.S. FTotal 33,093.5 119Extenders at 5° C. 702.95 5 140.59 2.2320.39 122. 911Extenders at 30° C. 6,418.55 5 1,283.7115,322.8Temperature 15,322.8 1Ejaculates 4,417.0 9Remainder (error) 6,232.2 99 62.95highly significant (P = < 0.01)TABLE 4. Analysis of Variance of thetozoa Remained Progressively MotileFactor S.S.Total 809,263Extenders at 5° C. 10,221.3Extenders at 30° C. 27,232Temperature---------337,345Ejaculates 175,099Remainder 259,365.7highly significant (P = < 0.01)Hours that 30 Per cent of the Sperma-VarianceD.F. F1075 2,044.35,446.45 1.85114. 47 1337,3451888 2,947Summary. The effect of terramycin, bacitracin, aureomycin, penicillin /streptomycin / sul fanilamide, and a combination of the aforementioned antibac-terial agents, on the motility of bovine spermatozoa and upon the control ofbacterial growth in bovine semen extended with citrate buffered yolk and storedat 5° C. and 30° C, was investigated. It was found that there was no signifi-cant difference in the motility of the spermatozoa or in the duration of a 3041Journal of the Arkansas Academy of Science, Vol. 6 [1953], Art. 11Published by Arkansas Academy of Science, 195342 ARKANSAS ACADEMY OF SCIENCEper cent motility rating of samples stored at 5° C that could be attributed todifferences in the action of extenders. A significant difference was found inthe percentage motility after 48 hours storage and the duration of motilitybetween samples stored at 5° C and 30° C, those stored at 30° C being consist-ently lower. Low motility ratings were observed in extenders containing baci-tracin, Lerramycin, and no antibacterial agent when samples were stored at 30°C. All antibacterial agents studied were highly bacteriostatic and/or bacteri-cidal with the exception of bacitracin when samples containing this antibioticwere stored at 30° C.References1. Almquist, J. 0., Thorp, W. T. S., and Knodt, C. B. The Effect of Penicillin upon theLivability, Glycolysis and Bacterial Content of Bovine Semen, volume 31, J. Dairy Sci.,pages 11-19; 1948.2. Almquist, J. 0., Glantz, P. J., and Thorp, W. T. S. The Effect of Streptomycin upon theLivability and Bacterial Content of Bovine Semen, volume 31, J. Dairy Sci., pages 501-507; 1948.3. Almquist, J. 0., Glantz, P. J., and Shaffer, H. E. The Effect of a Combination of Peni-cillin and Streptomycin upon the Livability and Bacterial Content of Bovine Semen, volume32, J. Dairy Sci., pages 183-190; 1949.4. Bush, L. J., Ludwick, T. M., Ferguson, L. C., and Ely, F. The Effect of Bacteria on theFertility of Bovine Semen, volume 33, J. Dairy Sci., pages 633-638; 1950.5. Finlay, A. C. ,Hobby, G. L., P'an, S. Y., Regna, P. P., Routien, J. B., Seeley, D. B.,Shull, G. M.,Sobin, B. A., Solomons, I. A., Vinson, J. W., and Kane, J. H. Terramycin, aNew Antibiotic, volume III,Science, page 85; 1950.6. Foote, R. H., and Bratton, R. W. Motility of Spermatozoa and Control of Bacteria inBovine Semen Extenders Containing Sulfanilamide, Polymyxin, and Aureomycin. volume 33, J.Dairy Sci. ,pages 539-543; 1950.7. Foote, R. H., and Bratton, R. W. Motility of Bovine Spermatozoa and Control of Bacteriaat 5° and 25* C. in Extenders Containing Sulfanilamide, Penicillin, Streptomycin and Poly-myxin. volume 33, J. Dairy Sci., pages 842-846; 1950.8. Foote, R. H. , and Salisbury, G. W. The Effect of Pyridium, Penicillin, Furacin, andPhenoxethol upon the Livability of Spermatozoa and upon the Control of Bacteria in Di-luted Bull Semen, volume 31, J. Dairy Sci., pages 763-768; 1948.9. Foote, R. H., and Salisbury, G. W. The Effect of Sulfonamides upon the Livability ofSpermatozoa and upon the Control of Bacteria in Diluted Bull Semen, volume 31, J. DairyScience, pages 769-778; 1948.10. Knodt, C. B., and Salisbury, G. W. The Effect of Sulfanilamide upon the Livability andMetabolism of Bovine Spermatozoa, volume 29, J. Dairy Sci., pages 285-291; 1946.11. Myers, R. M., Almquist, J. 0., and Prince, P. W. The Influence of Aureomycin upon theLivability and Bacterial Content of Bull Semen, volume 33, J. Dairy Sci., page 394; 1950.12. Olds, D., Oliver, L., and Seath, D. M. The Effect of Antibiotics on the Fertility of BullSemen and Their Relationship to the Estrus Cycle Length of Dairy Cattle Following Arti-ficial Insemination, volume 34, J. Dairy Sci., pages 966-970; 1951.13. Phillips, P. H., and Spitzer, R. R. -4 Snythetic Pablum for the Preservation of BullSemen, volume 29, J. Dairy Sci., pages 407-414; 1946.14. Rose, H. T. The Use of Bacitracin in Small Animal Medicine, volume 117, J. Am. Vet. Med.Ass'n., pages 306-310; 1950.15. Salisbury, G. W. , Willett, E. L., and Gunsalus, I. C. Some Problems in Bull Semen Stor-age, volume 32, Proc. Am. Soc. Animal Production, pages 210-212; 1939.16. Salisbury, G. W., and Knodt, C. B. The Effect of Sulfanilamide in the Diluent upon Fer-tility of Bull Semen, volume 30, J. Dairy Sci., pages 361-369; 1947.17. Snedecor, G. W. Statistical Methods, edition 4, The Iowa State College Press, Ames.;1946.18. Sykes, J. G., and Mixner, J. P. Toxicity to Bull Spermatozoa of Various Salts, Brands andLots of Penicillin, Streptomycin, Aureomycin and Chloromycetin. volume 34, J. Dairy Sci.,pages 342-46; 1951.42Journal of the Arkansas Academy of Science, Vol. 6 [1953], Art. 11https://scholarworks.uark.edu/jaas/vol6/iss1/11

Motility of Spermatazoa and Control of Bacteria in Bovine Semen Extenders Containing Sulfanilamide, Aureomycin, Terramycin, Bacitracin, Penicillin, and Strepromycin

ScholarWorks@UARK

Journal of the Arkansas Academy of ScienceVolume 6 Article 111953Motility of Spermatazoa and Control of Bacteria inBovine Semen Extenders ContainingSulfanilamide, Aureomycin, Terramycin, Bacitracin,Penicillin, and StrepromycinH. K. McCartneyUniversity of Arkansas, FayettevilleO. T. StallcupUniversity of Arkansas, FayettevilleFollow this and additional works at: http://scholarworks.uark.edu/jaasPart of the Other Animal Sciences CommonsThis article is available for use under the Creative Commons license: Attribution-NoDerivatives 4.0 International (CC BY-ND 4.0). Users are able toread, download, copy, print, distribute, search, link to the full texts of these articles, or use them for any other lawful purpose, without asking priorpermission from the publisher or the author.This Article is brought to you for free and open access by ScholarWorks@UARK. It has been accepted for inclusion in Journal of the Arkansas Academyof Science by an authorized editor of ScholarWorks@UARK. For more information, please contact scholar@uark.edu, ccmiddle@uark.edu.Recommended CitationMcCartney, H. K. and Stallcup, O. T. (1953) "Motility of Spermatazoa and Control of Bacteria in Bovine Semen Extenders ContainingSulfanilamide, Aureomycin, Terramycin, Bacitracin, Penicillin, and Strepromycin," Journal of the Arkansas Academy of Science: Vol. 6 ,Article 11.Available at: http://scholarworks.uark.edu/jaas/vol6/iss1/11MOTILITY OF SPERMATOZOA AND CONTROL OF BACTERIAIN BOVINE SEMEN EXTENDERS CONTAINING SULFANILAMIDE,AUREOMYCIN, TERRAMYCIN. BACITRACIN, PENICILLIN, AND STREPTOMYCINH. K. McCARTNEY and 0. T. STALLCUP 1University of ArkansasThe importance of controlling bacterial populations in bovine semen usedfor artificial insemination has been pointed out by Salisbury et al. (15). Asignificant relationship has been found between the number of bacteria in di-luted semen and its fertility when all types of bacteria were considered (4).The growth of many organisms commonly found in bovine semen is reportedly in-hibited by the addition of sulfonamides (6, 9, 10, 13, 16), of penicillin (1, 8,9, 18), of streptomycin (2, 18), of streptomycin plus penicillin (3, 12), ofpolymyxins (6), and of aureomycin (6, 11, 18), without significant toxic effectson. the motility of spermatozoa. However, no data have been reported on the con-trol of bacterial growth in semen by the use of bacitracin or terramycin.Finlay et al. (5) have reported that terramycin is bacteriostatic to manyGram-negative enteric organisms, aerobic spore formers, and Gram-positive cocci.It also has antirickettsial activity and appears to inhibit certain influenzaviruses.Rose (14) has reported that bacitracin is effective against most strains ofhemolytic streptococci, nonhemolytic streptococci, coagulase positive staphly-ococci, pneumococci , gonococci, anaerobic cocci in general, certain bacilli,certain spirochetes, actinomycetes and some protozoa. It shows little actionagainst the large group of aerobic Gram-negative nonsporeforming bacilli. Inview of the reported high bactericidal activity of terramycin and bacitracin, itseemed desirable to study the control of bacterial growth and effects on mo-tility of the spermatozoa by these compounds as compared to previously usedbactericidal agents.Materials and Methods. The antibacterial agents used in these investiga-tions are shown in Table 1.TAELE 1. Levels of Antibacterial Agents Used per 100 ml of Citrate-yolk ExtenderLPeni- Aureo-< i11in mycin Dihydro- TerramycinSulfa- G hydrochol- Bacitracin strepto- hydrochol-nilamide sodium ride (55 units mycin rideExtender (mgm) (mgn) (mgm) per mgn) (mgm) (mgn)Control ----0 0 0 0 0 0A 300 30 10 10 100 10BC-- --10D --.300 30- --100E--10Extender D was used for comparison because of the common usage of theseantibacterial agents in extenders. Extender A was used to determine ifa combi-nation of all the antibiotics studies was desirable. To make a simultaneouscomparison of the effects of the different antibiotics on the motility of sperm-The authors are indebted to Herbert G. Luther, Charles Pfizer and Company, Incorporation,for the terramycin hydrochloride used in these investigations, and to Gertrude M. Cox, direc-tor, Institute of Statistics, University of North Carolina, for assistance in outlining some ofthe statistical procedures used in the analysis of the data.Research Paper 1070 Journal Series. University of Arkansas*39Journal of the Arkansas Academy of Science, Vol. 6 [1953], Art. 11Published by Arkansas Academy of Science, 195340 ARKANSAS ACADEMY OF SCIENCEatozoa and upon bacterial control, semen samples obtained from the ArkansasArtificialBreeders' Association of Fayetteville, Arkansas, were divided intoas many equal portions as there were antibacterial agents to be tested. Eachportion of semen was mixed with 3 per cent citrate-yolk extender to which theantibiotics, or antibiotics plus sul fani 1amide, had been added. The averagedilution rate was one part of semen to 40 parts of citrate-yolk extender. Theextended semen then was divided into equal portions and placed in 9-ml tgsttubes filled to capacity. One portion was stored in a refrigerator at 5 Cwhile the other portion was exposed to a temperature of 30° C ± 2° C. Dailymicroscopic examinations of the diluted semen afforded an estimate of the ef-fects of the antibacterial agents on the motility of the spermatozoa. Averagemotility readings, after 48 hours of storage, are presented as this time in-terval and probably represent a near maximum storage time during which semen isused by most artificial breeding associations in this country. The samples wereread until the motility reading fell below 30 per cent and then discarded.Statistical significance between means was tested by analysis of variance (17).The number of living bacteria was determined by the plate count method.After incubation at 37° C for 48 hours, the number of colonies on each platewas counted and the number of bacteria that would have been present in an en-tire milliliterof the diluted semen was calculated. Control plates were madeto check the sterility of the water used for dilutions, of the agar, and of theatmosphere in the laboratory during the plating procedure.Results. A summary of the data concerning the spermatozoa motility, theduration of motility, and the number of bacteria in extended semen containingthe antibacterial agents studied is presented in Table 2. After 48 hours ofstorage at 5* C, bacterial growth was not excessive in any of the extended semensamples. The combination of all antibacterial agents usually inhibited bacteri-al growth completely. Bacitracin and aureomycin completely inhibited bacterialgrowth in 50 per cent of the samples, this difference in bactericidal actionapparently resulting from the predominating types of bacteria in differentsamples.TABLE 2. Motility of Spermatozoa, Duration of Motility, and Numbers of Bacteriain Extended Semen Containing Various Antibacterial AgentsDurationStorage After 48 hours storage motilityTemper- motile bacteria rating ofatures spermatozoa 1 per ml. 2 30 per cent 2°C per cent Thousands HoursControl 5° 39.8 5.0 145.330* 9.6 415.7 28.0A (Combination of anti- 5° 48.7 .3 181.3bacterial agents) 30° 32.8 2.5 85.3B (Terramycin hydrochlo- 5° 46.0 6.5 154.7ride) 30° 19.9 5.2 42.7C (Bacitracin) 5° 48.1 .8 180.030° 7.9 474.7 38.0D (Penicillin / strepto- 5° 44.1 5.1 173.3mycin / sulfanilamide) 30° 33.5 15.2 77.3E (Aureomycin hydrochlo- 5° 40.6 .7 156.0ride) 30* 28.0 18.2 58.7'Average of 10 ejaculates.Average of nine ejaculatesAt 30° C bacteria multiplied rapidly when no antibacterial agent was pres-ent as was the case when bacitracin was present, this antibacterial agent ap-parently being inactivated at the higher temperature. Putrefaction of the eggyolk consistently occurred when bacterial growth was high. The percentage of40Journal of the Arkansas Academy of Science, Vol. 6 [1953], Art. 11http://scholarworks.uark.edu/jaas/vol6/iss1/11MOTILITY OF BOVINE SPERMATOZOA 41samples exhibiting a putrid odor after 48 hour's storage at 30° C was 40, and30, respectively, for the extenders containing bacitracin and no antibacterialagents, respectively.The analyses of variance of the percentage of progressively motile sperma-tozoa after 48 hour's storage is presented in Table 3. At 5° C the percentageof motile spermatozoa was similar in all extenders throughout the 48 hour'sstorage period. The effects of temperature appear to be significant, all sam-ples showing a lower average motility after 48 hours at 30* C as compared to 5°C, regardless of the diluter used (P<0.01). Similar results for a 72 hourstorage interval were reported by Foote and Bratton (7). At 30° C after a 48hour storage period significant differences between extenders were observed(P<0.01). Aureomycin, penicillin / streptomycin / sul fanilamide, and a combi-nation of antibacterial agencs were significantly superior (P<0.01) to noantibacterial agent and bacitracin at this temperature. Bacitracin, terramycin,and no antibacterial agent were not significantly different from each other(P<0.05). This was probably caused by a high bacterial count in samples con-taining bacitracin, and the low motility of the samples containing terramycinmay have resulted from toxicity although no such effects were evident when sam-ples were stored at 5° C.An analysis of variance of the data concerning the length of time a pro-gressive motility rating of 30 per cent was maintained is presented in Table 4.The temperature effects were again significant, the duration of motility beingappreciably lower in samples stored at 30* C as compared to those stored at 5°C. The fact that the extender effect at 30* C is riot significant suggests thatthe criterion of response used here is not as precise as the one used in Table3. This is probably because the spermatozoa died rapidly in some extendersstored at 30° C, and daily microscopic examination would not detect preciselywhen 30 per cent of the spermatozoa were motile.TABLE 3. Analysis of Variance of Percentages of Progressively MotileSpermatozoa after 48 hour's StorageFactor S.S. D.F. M.S. FTotal 33,093.5 119Extenders at 5° C. 702.95 5 140.59 2.2320.39 122. 911Extenders at 30° C. 6,418.55 5 1,283.7115,322.8Temperature 15,322.8 1Ejaculates 4,417.0 9Remainder (error) 6,232.2 99 62.95highly significant (P = < 0.01)TABLE 4. Analysis of Variance of thetozoa Remained Progressively MotileFactor S.S.Total 809,263Extenders at 5° C. 10,221.3Extenders at 30° C. 27,232Temperature --------- 337,345Ejaculates 175,099Remainder 259,365.7highly significant (P = < 0.01)Hours that 30 Per cent of the Sperma-VarianceD.F. F1075 2,044.35,446.45 1.85114. 47 1337,3451888 2,947Summary. The effect of terramycin, bacitracin, aureomycin, penicillin /streptomycin / sul fanilamide, and a combination of the aforementioned antibac-terial agents, on the motility of bovine spermatozoa and upon the control ofbacterial growth in bovine semen extended with citrate buffered yolk and storedat 5° C. and 30° C, was investigated. It was found that there was no signifi-cant difference in the motility of the spermatozoa or in the duration of a 3041Journal of the Arkansas Academy of Science, Vol. 6 [1953], Art. 11Published by Arkansas Academy of Science, 195342 ARKANSAS ACADEMY OF SCIENCEper cent motility rating of samples stored at 5° C that could be attributed todifferences in the action of extenders. A significant difference was found inthe percentage motility after 48 hours storage and the duration of motilitybetween samples stored at 5° C and 30° C, those stored at 30° C being consist-ently lower. Low motility ratings were observed in extenders containing baci-tracin, Lerramycin, and no antibacterial agent when samples were stored at 30°C. All antibacterial agents studied were highly bacteriostatic and/or bacteri-cidal with the exception of bacitracin when samples containing this antibioticwere stored at 30° C.References1. Almquist, J. 0., Thorp, W. T. S., and Knodt, C. B. The Effect of Penicillin upon theLivability, Glycolysis and Bacterial Content of Bovine Semen, volume 31, J. Dairy Sci.,pages 11-19; 1948.2. Almquist, J. 0., Glantz, P. J., and Thorp, W. T. S. The Effect of Streptomycin upon theLivability and Bacterial Content of Bovine Semen, volume 31, J. Dairy Sci., pages 501-507; 1948.3. Almquist, J. 0., Glantz, P. J., and Shaffer, H. E. The Effect of a Combination of Peni-cillin and Streptomycin upon the Livability and Bacterial Content of Bovine Semen, volume32, J. Dairy Sci., pages 183-190; 1949.4. Bush, L. J., Ludwick, T. M., Ferguson, L. C., and Ely, F. The Effect of Bacteria on theFertility of Bovine Semen, volume 33, J. Dairy Sci., pages 633-638; 1950.5. Finlay, A. C. ,Hobby, G. L., P'an, S. Y., Regna, P. P., Routien, J. B., Seeley, D. B.,Shull, G. M.,Sobin, B. A., Solomons, I. A., Vinson, J. W., and Kane, J. H. Terramycin, aNew Antibiotic, volume III,Science, page 85; 1950.6. Foote, R. H., and Bratton, R. W. Motility of Spermatozoa and Control of Bacteria inBovine Semen Extenders Containing Sulfanilamide, Polymyxin, and Aureomycin. volume 33, J.Dairy Sci. ,pages 539-543; 1950.7. Foote, R. H., and Bratton, R. W. Motility of Bovine Spermatozoa and Control of Bacteriaat 5° and 25* C. in Extenders Containing Sulfanilamide, Penicillin, Streptomycin and Poly-myxin. volume 33, J. Dairy Sci., pages 842-846; 1950.8. Foote, R. H. , and Salisbury, G. W. The Effect of Pyridium, Penicillin, Furacin, andPhenoxethol upon the Livability of Spermatozoa and upon the Control of Bacteria in Di-luted Bull Semen, volume 31, J. Dairy Sci., pages 763-768; 1948.9. Foote, R. H., and Salisbury, G. W. The Effect of Sulfonamides upon the Livability ofSpermatozoa and upon the Control of Bacteria in Diluted Bull Semen, volume 31, J. DairyScience, pages 769-778; 1948.10. Knodt, C. B., and Salisbury, G. W. The Effect of Sulfanilamide upon the Livability andMetabolism of Bovine Spermatozoa, volume 29, J. Dairy Sci., pages 285-291; 1946.11. Myers, R. M., Almquist, J. 0., and Prince, P. W. The Influence of Aureomycin upon theLivability and Bacterial Content of Bull Semen, volume 33, J. Dairy Sci., page 394; 1950.12. Olds, D., Oliver, L., and Seath, D. M. The Effect of Antibiotics on the Fertility of BullSemen and Their Relationship to the Estrus Cycle Length of Dairy Cattle Following Arti-ficial Insemination, volume 34, J. Dairy Sci., pages 966-970; 1951.13. Phillips, P. H., and Spitzer, R. R. -4 Snythetic Pablum for the Preservation of BullSemen, volume 29, J. Dairy Sci., pages 407-414; 1946.14. Rose, H. T. The Use of Bacitracin in Small Animal Medicine, volume 117, J. Am. Vet. Med.Ass'n., pages 306-310; 1950.15. Salisbury, G. W. , Willett, E. L., and Gunsalus, I. C. Some Problems in Bull Semen Stor-age, volume 32, Proc. Am. Soc. Animal Production, pages 210-212; 1939.16. Salisbury, G. W., and Knodt, C. B. The Effect of Sulfanilamide in the Diluent upon Fer-tility of Bull Semen, volume 30, J. Dairy Sci., pages 361-369; 1947.17. Snedecor, G. W. Statistical Methods, edition 4, The Iowa State College Press, Ames.;1946.18. Sykes, J. G., and Mixner, J. P. Toxicity to Bull Spermatozoa of Various Salts, Brands andLots of Penicillin, Streptomycin, Aureomycin and Chloromycetin. volume 34, J. Dairy Sci.,pages 342-46; 1951.42Journal of the Arkansas Academy of Science, Vol. 6 [1953], Art. 11http://scholarworks.uark.edu/jaas/vol6/iss1/11

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Motility of Spermatazoa and Control of Bacteria in Bovine Semen Extenders Containing Sulfanilamide, Aureomycin, Terramycin, Bacitracin, Penicillin, and Strepromycin

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